Bio

Clinical Focus


  • Pathology
  • Pathology and Laboratory Medicine

Academic Appointments


Administrative Appointments


  • Chair, Stanford University School of Medicine - Pathology (1999 - Present)

Professional Education


  • Fellowship:Massachusetts General Hospital (1979) MA
  • Medical Education:Harvard Medical School (1973) MA
  • Residency:Massachusetts General Hospital (1977) MA

Research & Scholarship

Current Research and Scholarly Interests


Mast cells, which normally reside in the tissues, and basophils, which circulate in the blood, are major effector cells of asthma and other IgE-associated allergic disorders and immune responses to parasites. However, mast cells also have been implicated (as effector and/or immunoregulatory cells) in many other settings, including certain autoimmune or inflammatory disorders, innate immune responses to pathogens and resistance to exogenous and exogenous agents which can express significant toxicity; mast cells also may contribute importantly, in certain settings, to angiogenesis, the regulation of epithelial development and function and fibrosis and other examples of tissue remodeling.

The goals of Dr. Galliĺs laboratory are to develop and employ genetic approaches in mice to understand the regulation of mast cell and basophil development and the expression of mast cell and basophil function, and to elucidate the roles of these cells in health and disease. In parallel with these mouse studies, we investigate the roles of mast cells in human health and disease by conducting studies of human mast cells, or by analyzing specimens derived from patients with asthma or other disorders in which mast cell have been implicated.

Teaching

2013-14 Courses


Publications

Journal Articles


  • Mast Cells Are Required for Full Expression of Allergen/SEB-Induced Skin Inflammation JOURNAL OF INVESTIGATIVE DERMATOLOGY Ando, T., Matsumoto, K., Namiranian, S., Yamashita, H., Glatthorn, H., Kimura, M., Dolan, B. R., Lee, J. J., Galli, S. J., Kawakami, Y., Jamora, C., Kawakami, T. 2013; 133 (12): 2695-2705

    Abstract

    Atopic dermatitis (AD) is a chronic pruritic inflammatory skin disease. We recently described an animal model in which repeated epicutaneous applications of a house dust mite extract and Staphylococcal enterotoxin B induced eczematous skin lesions. In this study we showed that global gene expression patterns are very similar between human AD skin and allergen/staphylococcal enterotoxin B-induced mouse skin lesions, particularly in the expression of genes related to epidermal growth/differentiation, skin barrier, lipid/energy metabolism, immune response, or extracellular matrix. In this model, mast cells and T cells, but not B cells or eosinophils, were shown to be required for the full expression of dermatitis, as revealed by reduced skin inflammation and reduced serum IgE levels in mice lacking mast cells or T cells (TCR?(-/-) or Rag1(-/-)). The clinical severity of dermatitis correlated with the numbers of mast cells, but not eosinophils. Consistent with the idea that T helper type 2 (Th2) cells play a predominant role in allergic diseases, the receptor for the Th2-promoting cytokine thymic stromal lymphopoietin and the high-affinity IgE receptor, Fc?RI, were required to attain maximal clinical scores. Therefore, this clinically relevant model provides mechanistic insights into the pathogenic mechanism of human AD.Journal of Investigative Dermatology advance online publication, 11 July 2013; doi:10.1038/jid.2013.250.

    View details for DOI 10.1038/jid.2013.250

    View details for Web of Science ID 000327015400010

    View details for PubMedID 23752044

  • Rapid desensitization induces internalization of antigen-specific IgE on mouse mast cells. journal of allergy and clinical immunology Oka, T., Rios, E. J., Tsai, M., Kalesnikoff, J., Galli, S. J. 2013; 132 (4): 922-932 e16

    Abstract

    Rapid desensitization transiently prevents severe allergic reactions, allowing administration of life-saving therapies in previously sensitized patients. However, the mechanisms underlying successful rapid desensitization are not fully understood.We sought to investigate whether the mast cell (MC) is an important target of rapid desensitization in mice sensitized to exhibit IgE-dependent passive systemic anaphylaxis inávivo and to investigate the antigen specificity and underlying mechanisms of rapid desensitization in our mouse model.C57BL/6 mice (inávivo) or primary isolated C57BL/6 mouse peritoneal mast cells (PMCs; inávitro) were passively sensitized with antigen-specific anti-2,4-dinitrophenyl IgE, anti-ovalbumin IgE, or both. MCs were exposed over a short period of time to increasing amounts of antigen (2,4-dinitrophenyl-human serum albumin or ovalbumin) in the presence of extracellular calcium inávitro or by means of intravenous administration to sensitized mice inávivo before challenging the mice with or exposing the PMCs to optimal amounts of specific or irrelevant antigen.Rapidly exposing mice or PMCs to progressively increasing amounts of specific antigen inhibited the development of antigen-induced hypothermia in sensitized mice inávivo and inhibited antigen-induced PMC degranulation and prostaglandin D2 synthesis inávitro. Such MC hyporesponsiveness was induced antigen-specifically and was associated with a significant reduction in antigen-specific IgE levels on MC surfaces.Rapidly exposing MCs to progressively increasingáamounts of antigen can both enhance the internalization of antigen-specific IgE on the MC surface andáalso desensitize these cells in an antigen-specific manneráinávivo and inávitro.

    View details for DOI 10.1016/j.jaci.2013.05.004

    View details for PubMedID 23810240

  • PLA2G3 promotes mast cell maturation and function. Nature immunology Starkl, P., Marichal, T., Galli, S. J. 2013; 14 (6): 527-529

    View details for DOI 10.1038/ni.2612

    View details for PubMedID 23685814

  • Mast cell anaphylatoxin receptor expression can enhance IgE-dependent skin inflammation in mice JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY Schaefer, B., Piliponsky, A. M., Oka, T., Song, C. H., Gerard, N. P., Gerard, C., Tsai, M., Kalesnikoff, J., Galli, S. J. 2013; 131 (2): 541-?

    Abstract

    Mast cells express receptors for complement anaphylatoxins C3a and C5a (ie, C3a receptor [C3aR] and C5a receptor [C5aR]), and C3a and C5a are generated during various IgE-dependent immediate hypersensitivity reactions in vivo. However, it is not clear to what extent mast cell expression of C3aR or C5aR influences C3a- or C5a-induced cutaneous responses or IgE-dependent mast cell activation and passive cutaneous anaphylaxis (PCA) in vivo.We sought to assess whether mouse skin mast cell expression of C3aR or C5aR influences (1) the cells' responsiveness to intradermal injections of C3a or C5a or (2) the extent of IgE-dependent mast cell degranulation and PCA in vivo.We measured the magnitude of cutaneous responses to intradermal injections of C3a or C5a and the extent of IgE-dependent mast cell degranulation and PCA responses in mice containing mast cells that did or did not express C3aR or C5aR.The majority of the skin swelling induced by means of intradermal injection of C3a or C5a required that mast cells at the site expressed C3aR or C5aR, respectively, and the extent of IgE-dependent degranulation of skin mast cells and IgE-dependent PCA was significantly reduced when mast cells lacked either C3aR or C5aR. IgE-dependent PCA responses associated with local increases in C3a levels occurred in antibody-deficient mice but not in mice deficient in Fc?RI?.Expression of C3aR and C5aR by skin mast cells contributes importantly to the ability of C3a and C5a to induce skin swelling and can enhance mast cell degranulation and inflammation during IgE-dependent PCA in vivo.

    View details for DOI 10.1016/j.jaci.2012.05.009

    View details for Web of Science ID 000314661500034

    View details for PubMedID 22728083

  • Evidence that mast cells are not required for healing of splinted cutaneous excisional wounds in mice. PloS one Nauta, A. C., Grova, M., Montoro, D. T., Zimmermann, A., Tsai, M., Gurtner, G. C., Galli, S. J., Longaker, M. T. 2013; 8 (3)

    Abstract

    Wound healing is a complex biological process involving the interaction of many cell types to replace lost or damaged tissue. Although the biology of wound healing has been extensively investigated, few studies have focused on the role of mast cells. In this study, we investigated the possible role of mast cells in wound healing by analyzing aspects of cutaneous excisional wound healing in three types of genetically mast cell-deficient mice. We found that C57BL/6-Kit(W-sh/W-sh), WBB6F1-Kit(W/W-v), and Cpa3-Cre; Mcl-1(fl/fl) mice re-epithelialized splinted excisional skin wounds at rates very similar to those in the corresponding wild type or control mice. Furthermore, at the time of closure, scars were similar in the genetically mast cell-deficient mice and the corresponding wild type or control mice in both quantity of collagen deposition and maturity of collagen fibers, as evaluated by Masson's Trichrome and Picro-Sirius red staining. These data indicate that mast cells do not play a significant non-redundant role in these features of the healing of splinted full thickness excisional cutaneous wounds in mice.

    View details for DOI 10.1371/journal.pone.0059167

    View details for PubMedID 23544053

  • New models for analyzing mast cell functions in vivo TRENDS IN IMMUNOLOGY Reber, L. L., Marichal, T., Galli, S. J. 2012; 33 (12): 613-625

    Abstract

    In addition to their well-accepted role as critical effector cells in anaphylaxis and other acute IgE-mediated allergic reactions, mast cells (MCs) have been implicated in a wide variety of processes that contribute to disease or help to maintain health. Although some of these roles were first suggested by analyses of MC products or functions in vitro, it is critical to determine whether, and under which circumstances, such potential roles actually can be performed by MCs in vivo. This review discusses recent advances in the development and analysis of mouse models to investigate the roles of MCs and MC-associated products during biological responses in vivo, and comments on some of the similarities and differences in the results obtained with these newer versus older models of MC deficiency.

    View details for DOI 10.1016/j.it.2012.09.008

    View details for Web of Science ID 000312055700005

    View details for PubMedID 23127755

  • Evidence questioning cromolyn's effectiveness and selectivity as a 'mast cell stabilizer' in mice LABORATORY INVESTIGATION Oka, T., Kalesnikoff, J., Starkl, P., Tsai, M., Galli, S. J. 2012; 92 (10): 1472-1482

    Abstract

    Cromolyn, widely characterized as a 'mast cell stabilizer', has been used in mice to investigate the biological roles of mast cells in vivo. However, it is not clear to what extent cromolyn can either limit the function of mouse mast cells or influence biological processes in mice independently of effects on mast cells. We confirmed that cromolyn (at 10 mg/kg in vivo or 10-100 ?M in vitro) can inhibit IgE-dependent mast cell activation in rats in vivo (measuring Evans blue extravasation in passive cutaneous anaphylaxis (PCA) and increases in plasma histamine in passive systemic anaphylaxis (PSA)) and in vitro (measuring peritoneal mast cell (PMC) ?-hexosaminidase release and prostaglandin D(2) synthesis). However, under the conditions tested, cromolyn did not inhibit those mast cell-dependent responses in mice. In mice, cromolyn also failed to inhibit the ear swelling or leukocyte infiltration at sites of PCA. Nor did cromolyn inhibit IgE-independent degranulation of mouse PMCs induced by various stimulators in vitro. At 100 mg/kg, a concentration 10 times higher than that which inhibited PSA in rats, cromolyn significantly inhibited the increases in plasma concentrations of mouse mast cell protease-1 (but not of histamine) during PSA, but had no effect on the reduction in body temperature in this setting. Moreover, this concentration of cromolyn (100 mg/kg) also inhibited LPS-induced TNF production in genetically mast cell-deficient C57BL/6-Kit(W-sh/W-sh) mice in vivo. These results question cromolyn's effectiveness and selectivity as an inhibitor of mast cell activation and mediator release in the mouse.

    View details for DOI 10.1038/labinvest.2012.116

    View details for Web of Science ID 000309324600009

    View details for PubMedID 22906983

  • The Chymase Mouse Mast Cell Protease 4 Degrades TNF, Limits Inflammation, and Promotes Survival in a Model of Sepsis AMERICAN JOURNAL OF PATHOLOGY Piliponsky, A. M., Chen, C., Rios, E. J., Treuting, P. M., Lahiri, A., Abrink, M., Pejler, G., Tsai, M., Galli, S. J. 2012; 181 (3): 875-886

    Abstract

    Mouse mast cell protease 4 (mMCP-4), the mouse counterpart of human mast cell chymase, is thought to have proinflammatory effects in innate or adaptive immune responses associated with mast cell activation. However, human chymase can degrade the proinflammatory cytokine TNF, a mediator that can be produced by mast cells and many other cell types. We found that mMCP-4 can reduce levels of mouse mast cell-derived TNF in vitro through degradation of transmembrane and soluble TNF. We assessed the effects of interactions between mMCP-4 and TNF in vivo by analyzing the features of a classic model of polymicrobial sepsis, cecal ligation and puncture (CLP), in C57BL/6J-mMCP-4-deficient mice versus C57BL/6J wild-type mice, and in C57BL/6J-Kit(W-sh/W-sh) mice containing adoptively transferred mast cells that were either wild type or lacked mMCP-4, TNF, or both mediators. The mMCP-4-deficient mice exhibited increased levels of intraperitoneal TNF, higher numbers of peritoneal neutrophils, and increased acute kidney injury after CLP, and also had significantly higher mortality after this procedure. Our findings support the conclusion that mMCP-4 can enhance survival after CLP at least in part by limiting detrimental effects of TNF, and suggest that mast cell chymase may represent an important negative regulator of TNF in vivo.

    View details for DOI 10.1016/j.ajpath.2012.05.013

    View details for Web of Science ID 000309251100016

    View details for PubMedID 22901752

  • Critical role of P1-Runx1 in mouse basophil development BLOOD Mukai, K., Benbarak, M. J., Tachibana, M., Nishida, K., Karasuyama, H., Taniuchi, I., Galli, S. J. 2012; 120 (1): 76-85

    Abstract

    Runx1(P1N/P1N) mice are deficient in the transcription factor distal promoter-derived Runt-related transcription factor 1 (P1-Runx1) and have a > 90% reduction in the numbers of basophils in the BM, spleen, and blood. In contrast, Runx1(P1N/P1N) mice have normal numbers of the other granulocytes (neutrophils and eosinophils). Although basophils and mast cells share some common features, Runx1(P1N/P1N) mice have normal numbers of mast cells in multiple tissues. Runx1(P1N/P1N) mice fail to develop a basophil-dependent reaction, IgE-mediated chronic allergic inflammation of the skin, but respond normally when tested for IgE- and mast cell-dependent passive cutaneous anaphylaxis in vivo or IgE-dependent mast cell degranulation in vitro. These results demonstrate that Runx1(P1N/P1N) mice exhibit markedly impaired function of basophils, but not mast cells. Infection with the parasite Strongyloides venezuelensis and injections of IL-3, each of which induces marked basophilia in wild-type mice, also induce modest expansions of the very small populations of basophils in Runx1(P1N/P1N) mice. Finally, Runx1(P1N/P1N) mice have normal numbers of the granulocyte progenitor cells, SN-Flk2(+/-), which can give rise to all granulocytes, but exhibit a > 95% reduction in basophil progenitors. The results of the present study suggest that P1-Runx1 is critical for a stage of basophil development between SN-Flk2(+/-) cells and basophil progenitors.

    View details for DOI 10.1182/blood-2011-12-399113

    View details for Web of Science ID 000307411100015

    View details for PubMedID 22611151

  • Modulation of mTOR Effector Phosphoproteins in Blood Basophils from Allergic Patients JOURNAL OF CLINICAL IMMUNOLOGY Gernez, Y., Tirouvanziam, R., Reshamwala, N., Yu, G., Weldon, B. C., Galli, S. J., Herzenberg, L. A., Nadeau, K. C. 2012; 32 (3): 565-573

    Abstract

    The mammalian target of rapamycin (mTOR) pathway contributes to various immunoinflammatory processes. Yet, its potential involvement in basophil responses in allergy remains unclear. In this pilot study, we quantified two key mTOR effector phosphoproteins, the eukaryotic initiation factor 4E (peIF4E) and S6 ribosomal protein (pS6rp), in blood basophils from nut allergy patients (NA, N?=?16) and healthy controls (HC, N?=?13). Without stimulation in vitro, basophil peIF4E levels were higher in NA than HC subjects (P?=?0.014). Stimulation with nut (offending) but not chicken / rice (non-offending) extract increased basophil peIF4E and pS6rp levels (+32%, P?=?0.018, and +98%, P?=?0.0026, respectively) in NA but not HC subjects, concomitant with increased surface levels of CD203c and CD63, both known to reflect basophil activation. Pre-treatment with the mTOR inhibitor rapamycin decreased pS6rp and CD203c responses in nut extract-stimulated basophils in NA subjects. Thus, basophil responses to offending allergens are associated with modulation of mTOR effector phosphoproteins.

    View details for DOI 10.1007/s10875-012-9651-x

    View details for Web of Science ID 000305982100019

    View details for PubMedID 22350221

  • IgE and mast cells in allergic disease NATURE MEDICINE Galli, S. J., Tsai, M. 2012; 18 (5): 693-704

    Abstract

    Immunoglobulin E (IgE) antibodies and mast cells have been so convincingly linked to the pathophysiology of anaphylaxis and other acute allergic reactions that it can be difficult to think of them in other contexts. However, a large body of evidence now suggests that both IgE and mast cells are also key drivers of the long-term pathophysiological changes and tissue remodeling associated with chronic allergic inflammation in asthma and other settings. Such potential roles include IgE-dependent regulation of mast-cell functions, actions of IgE that are largely independent of mast cells and roles of mast cells that do not directly involve IgE. In this review, we discuss findings supporting the conclusion that IgE and mast cells can have both interdependent and independent roles in the complex immune responses that manifest clinically as asthma and other allergic disorders.

    View details for DOI 10.1038/nm.2755

    View details for Web of Science ID 000303763500037

    View details for PubMedID 22561833

  • Reduced mast cell and basophil numbers and function in Cpa3-Cre; Mcl-1(fl/fl) mice BLOOD Lilla, J. N., Chen, C., Mukai, K., Benbarak, M. J., Franco, C. B., Kalesnikoff, J., Yu, M., Tsai, M., Piliponsky, A. M., Galli, S. J. 2011; 118 (26): 6930-6938

    Abstract

    It has been reported that the intracellular antiapoptotic factor myeloid cell leukemia sequence 1 (Mcl-1) is required for mast cell survival in vitro, and that genetic manipulation of Mcl-1 can be used to delete individual hematopoietic cell populations in vivo. In the present study, we report the generation of C57BL/6 mice in which Cre recombinase is expressed under the control of a segment of the carboxypeptidase A3 (Cpa3) promoter. C57BL/6-Cpa3-Cre; Mcl-1(fl/fl) mice are severely deficient in mast cells (92%-100% reduced in various tissues analyzed) and also have a marked deficiency in basophils (58%-78% reduced in the compartments analyzed), whereas the numbers of other hematopoietic cell populations exhibit little or no changes. Moreover, Cpa3-Cre; Mcl-1(fl/fl) mice exhibited marked reductions in the tissue swelling and leukocyte infiltration that are associated with both mast cell- and IgE-dependent passive cutaneous anaphylaxis (except at sites engrafted with in vitro-derived mast cells) and a basophil- and IgE-dependent model of chronic allergic inflammation, and do not develop IgE-dependent passive systemic anaphylaxis. Our findings support the conclusion that Mcl-1 is required for normal mast cell and basophil development/survival in vivo in mice, and also suggest that Cpa3-Cre; Mcl-1(fl/fl) mice may be useful in analyzing the roles of mast cells and basophils in health and disease.

    View details for DOI 10.1182/blood-2011-03-343962

    View details for Web of Science ID 000298401000030

    View details for PubMedID 22001390

  • Phenotypic and functional plasticity of cells of innate immunity: macrophages, mast cells and neutrophils NATURE IMMUNOLOGY Galli, S. J., Borregaard, N., Wynn, T. A. 2011; 12 (11): 1035-1044

    Abstract

    Hematopoietic cells, including lymphoid and myeloid cells, can develop into phenotypically distinct 'subpopulations' with different functions. However, evidence indicates that some of these subpopulations can manifest substantial plasticity (that is, undergo changes in their phenotype and function). Here we focus on the occurrence of phenotypically distinct subpopulations in three lineages of myeloid cells with important roles in innate and acquired immunity: macrophages, mast cells and neutrophils. Cytokine signals, epigenetic modifications and other microenvironmental factors can substantially and, in some cases, rapidly and reversibly alter the phenotype of these cells and influence their function. This suggests that regulation of the phenotype and function of differentiated hematopoietic cells by microenvironmental factors, including those generated during immune responses, represents a common mechanism for modulating innate or adaptive immunity.

    View details for DOI 10.1038/ni.2109

    View details for Web of Science ID 000296500100008

    View details for PubMedID 22012443

  • Mast cell chymase reduces the toxicity of Gila monster venom, scorpion venom, and vasoactive intestinal polypeptide in mice JOURNAL OF CLINICAL INVESTIGATION Akahoshi, M., Song, C. H., Piliponsky, A. M., Metz, M., Guzzetta, A., Abrink, M., Schlenner, S. M., Feyerabend, T. B., Rodewald, H., Pejler, G., Tsai, M., Galli, S. J. 2011; 121 (10): 4180-4191

    Abstract

    Mast cell degranulation is important in the pathogenesis of anaphylaxis and allergic disorders. Many animal venoms contain components that can induce mast cell degranulation, and this has been thought to contribute to the pathology and mortality caused by envenomation. However, we recently reported evidence that mast cells can enhance the resistance of mice to the venoms of certain snakes and that mouse mast cell-derived carboxypeptidase A3 (CPA3) can contribute to this effect. Here, we investigated whether mast cells can enhance resistance to the venom of the Gila monster, a toxic component of that venom (helodermin), and the structurally similar mammalian peptide, vasoactive intestinal polypeptide (VIP). Using 2 types of mast cell-deficient mice, as well as mice selectively lacking CPA3 activity or the chymase mouse mast cell protease-4 (MCPT4), we found that mast cells and MCPT4, which can degrade helodermin, can enhance host resistance to the toxicity of Gila monster venom. Mast cells and MCPT4 also can limit the toxicity associated with high concentrations of VIP and can reduce the morbidity and mortality induced by venoms from 2 species of scorpions. Our findings support the notion that mast cells can enhance innate defense by degradation of diverse animal toxins and that release of MCPT4, in addition to CPA3, can contribute to this mast cell function.

    View details for DOI 10.1172/JCI46139

    View details for Web of Science ID 000295601000043

    View details for PubMedID 21926462

  • Identification of an IFN-gamma/mast cell axis in a mouse model of chronic asthma JOURNAL OF CLINICAL INVESTIGATION Yu, M., Eckart, M. R., Morgan, A. A., Mukai, K., Butte, A. J., Tsai, M., Galli, S. J. 2011; 121 (8): 3133-3143

    Abstract

    Asthma is considered a Th2 cell?associated disorder. Despite this, both the Th1 cell?associated cytokine IFN-? and airway neutrophilia have been implicated in severe asthma. To investigate the relative contributions of different immune system components to the pathogenesis of asthma, we previously developed a model that exhibits several features of severe asthma in humans, including airway neutrophilia and increased lung IFN-?. In the present studies, we tested the hypothesis that IFN-? regulates mast cell function in our model of chronic asthma. Engraftment of mast cell?deficient KitW(-sh/W-sh) mice, which develop markedly attenuated features of disease, with wild-type mast cells restored disease pathology in this model of chronic asthma. However, disease pathology was not fully restored by engraftment with either IFN-? receptor 1?null (Ifngr1?/?) or Fc? receptor 1??null (Fcer1g?/?) mast cells. Additional analysis, including gene array studies, showed that mast cell expression of IFN-?R contributed to the development of many Fc?RI?-dependent and some Fc?RI?-independent features of disease in our model, including airway hyperresponsiveness, neutrophilic and eosinophilic inflammation, airway remodeling, and lung expression of several cytokines, chemokines, and markers of an alternatively activated macrophage response. These findings identify a previously unsuspected IFN-?/mast cell axis in the pathology of chronic allergic inflammation of the airways in mice.

    View details for DOI 10.1172/JCI43598

    View details for Web of Science ID 000293495500024

    View details for PubMedID 21737883

  • Basophil CD203c Levels Are Increased at Baseline and Can Be Used to Monitor Omalizumab Treatment in Subjects with Nut Allergy INTERNATIONAL ARCHIVES OF ALLERGY AND IMMUNOLOGY Gernez, Y., Tirouvanziam, R., Yu, G., Ghosn, E. E., Reshamwala, N., Tammie Nguyen, T., Tsai, M., Galli, S. J., Herzenberg, L. A., Herzenberg, L. A., Nadeau, K. C. 2011; 154 (4): 318-327

    Abstract

    Basophils contribute to anaphylaxis and allergies. We examined the utility of assessing basophil-associated surface antigens (CD11b/CD63/CD123/CD203c/CD294) in characterizing and monitoring subjects with nut allergy.We used flow cytometry to analyze basophils at baseline (without any activation) and after ex vivo stimulation of whole blood by addition of nut or other allergens for 2, 10, and 30 min. We also evaluated whether basophil expression of CD11b/CD63/CD123/CD203c/CD294 was altered in subjects treated with anti-IgE monoclonal antibody (omalizumab) to reduce plasma levels of IgE.We demonstrate that basophil CD203c levels are increased at baseline in subjects with nut allergy compared to healthy controls (13 subjects in each group, p < 0.0001). Furthermore, we confirm that significantly increased expression of CD203c occurs on subject basophils when stimulated with the allergen to which the subject is sensitive and can be detected rapidly (10 min of stimulation, n = 11, p < 0.0008). In 5 subjects with severe peanut allergy, basophil CD203c expression following stimulation with peanut allergen was significantly decreased (p < 0.05) after 4 and 8 weeks of omalizumab treatment but returned toward pretreatment levels after treatment cessation.Subjects with nut allergy show an increase of basophil CD203c levels at baseline and following rapid ex vivo stimulation with nut allergen. Both can be reduced by omalizumab therapy. These results highlight the potential of using basophil CD203c levels for baseline diagnosis and therapeutic monitoring in subjects with nut allergy.

    View details for DOI 10.1159/000321824

    View details for Web of Science ID 000288529200007

    View details for PubMedID 20975283

  • MAST CELLS AND IMMUNOREGULATION/IMMUNOMODULATION MAST CELL BIOLOGY: CONTEMPORARY AND EMERGING TOPICS Tsai, M., Grimbaldeston, M., Galli, S. J. 2011; 716: 186-211

    Abstract

    Mast cells often represent one of the first cells of the immune system to interact with environmental antigens, invading pathogens or environmentally-derived toxins. Mast cells also can undergo alterations in phenotype, anatomic distribution and numbers during innate or adaptive immune responses. In addition to their well-known roles as effector cells during IgE- and antigen-induced allergic reactions, mast cells can be activated by many other signals, including some that are derived directly from pathogens or which are generated during innate or adaptive immune responses. Mast cells also express many costimulatory molecules with immunoregulatory activities and can secrete many products that can positively or negatively regulate immune responses. In this chapter, we describe mouse models used for analyzing mast-cell function in vivo and illustrate how such models have been used to identify positive or negative immunomodulatory roles for mast cells during specific innate or adaptive immune responses. We also briefly describe some of the mast-cell functions, products and surface receptors that have the potential to permit mast cells to promote or suppress immune responses that can either enhance host defense or contribute to disease.

    View details for Web of Science ID 000290066600011

    View details for PubMedID 21713658

  • Antiinflammatory and immunosuppressive functions of mast cells. Methods in molecular biology (Clifton, N.J.) Kalesnikoff, J., Galli, S. J. 2011; 677: 207-220

    Abstract

    Through the release of biologically active products, mast cells function as important effector and immunoregulatory cells in diverse immunological reactions and other biological responses; for example, mast cells promote inflammation and other tissue changes in immunoglobulin E (IgE)-associated allergic disorders, as well as in certain innate and adaptive immune responses that are thought to be independent of IgE. Despite the mast cell's well-deserved reputation as a promoter of inflammation, others and we have used bone marrow-derived cultured mast cell (BMCMC) engrafted mast cell-deficient c-kit-mutant mice (so-called "mast cell knock-in" mice) to show that mast cells can also have important antiinflammatory and immunosuppressive functions in vivo. An early study showed that mast cells can contribute to susceptibility to ultraviolet B (UVB)-induced immunosuppression in one model of contact hypersensitivity (CHS), through effects mediated at least in part by histamine. Subsequently, it was reported that mast cells can mediate negative immunomodulatory effects following Anopheles mosquito bites, and in peripheral tolerance to skin allografts; however, the mechanism(s) by which mast cells mediate immunosuppressive functions in these two studies remains to be elucidated. Finally, we showed that mast cells and mast cell-derived IL-10 can limit the magnitude of and promote the resolution of certain CHS responses, and suppress the inflammation and skin injury associated with innate cutaneous responses to chronic low-dose UVB irradiation. This chapter outlines the generation of BMCMCs, a powerful model system commonly used to: (1) identify potential mast cell mediators in vitro; (2) study the mechanisms of mast cell activation and mediator release in response to specific stimuli in vitro; and (3) engraft mast cell-deficient mice to study the effector and immunoregulatory roles of mast cells or specific mast cell mediators in diverse immunological responses in vivo.

    View details for DOI 10.1007/978-1-60761-869-0_15

    View details for PubMedID 20941613

  • Thymic Stromal Lymphopoietin Contributes to Myeloid Hyperplasia and Increased Immunoglobulins, But Not Epidermal Hyperplasia, in RabGEF1-Deficient Mice AMERICAN JOURNAL OF PATHOLOGY Tsai, M., Chen, C., Mukai, K., Song, C. H., Thompson, L. J., Ziegler, S. F., Tam, S., Galli, S. J. 2010; 177 (5): 2411-2420

    Abstract

    Mice overexpressing the proallergic cytokine thymic stromal lymphopoietin (TSLP) in the skin develop a pathology resembling atopic dermatitis. RabGEF1, a guanine nucleotide exchange factor for Rab5 GTPase, is a negative regulator of IgE-dependent mast cell activation, and Rabgef1-/- and TSLP transgenic mice share many similar phenotypic characteristics, including elevated serum IgE levels and severe skin inflammation, with infiltrates of both lymphocytes and eosinophils. We report here that Rabgef1-/- mice also develop splenomegaly, lymphadenopathy, myeloid hyperplasia, and high levels of TSLP. Rabgef1-/-TSLPR-/- mice, which lack TSLP/TSLP receptor (TSLPR) signaling, had levels of blood neutrophils, spleen myeloid cells, and serum IL-4, IgG1, and IgE levels that were significantly reduced compared with those in Rabgef1-/-TSLPR+/+ mice. However, Rabgef1-/-TSLPR-/- mice, like Rag1- or eosinophil-deficient Rabgef1-/- mice, developed cutaneous inflammation and epidermal hyperplasia. Therefore, in Rabgef1-/- mice, TSLP/TSLPR interactions are not required for the development of epidermal hyperplasia but contribute to the striking myeloid hyperplasia and overproduction of immunoglobulins observed in these animals. Our study shows that RabGEF1 can negatively regulate TSLP production in vivo and that excessive production of TSLP contributes to many of the phenotypic abnormalities in Rabgef1-/- mice. However, the marked epidermal hyperplasia, cutaneous inflammation, and increased numbers of dermal mast cells associated with RabGEF1 deficiency can develop via a TSLPR-independent pathway, as well as in the absence of Rag1 or eosinophils.

    View details for DOI 10.2353/ajpath.2010.100181

    View details for Web of Science ID 000284182900028

    View details for PubMedID 20829437

  • Mast cells in allergy and infection: Versatile effector and regulatory cells in innate and adaptive immunity EUROPEAN JOURNAL OF IMMUNOLOGY Galli, S. J., Tsai, M. 2010; 40 (7): 1843-1851

    Abstract

    Mast cells are widely distributed in tissues, particularly near surfaces exposed to the environment. Mast cells can be activated to secrete diverse mediators and cytokines by IgE and specific Ag and many other stimuli, including products derived from either pathogens or the host during innate immune responses. Although mast cells are best known for their role in IgE-associated allergic disorders, mast cells can also exacerbate models of autoimmunity, enhance the sensitization and/or effector phases of certain cutaneous contact hypersensitivity responses, and increase inflammation and mortality during some severe bacterial infections. In other settings, however, mast cells can limit inflammation and tissue injury: mast cells promote host resistance in certain models of bacterial or parasite infection, limit pathology during some acquired immune responses to environmental Ag, including examples of severe contact hypersensitivity, and have adjuvant-like properties that can enhance the development of protective immunity against pathogens. These and other findings suggest that mast cells occupy a critical niche at the interface of innate and acquired immunity, where, depending on circumstances that remain to be fully understood, mast cells may function to perturb or help to restore homeostasis (or both), with consequences that can either promote health or contribute to disease.

    View details for DOI 10.1002/eji.201040559

    View details for Web of Science ID 000280220600012

    View details for PubMedID 20583030

  • Regulation of secretory granule size by the precise generation and fusion of unit granules JOURNAL OF CELLULAR AND MOLECULAR MEDICINE Hammel, I., Lagunoff, D., Galli, S. J. 2010; 14 (7): 1904-1916

    Abstract

    Morphometric evidence derived from studies of mast cells, pancreatic acinar cells and other cell types supports a model in which the post-Golgi processes that generate mature secretory granules can be resolved into three steps: (1) fusion of small, Golgi-derived progranules to produce immature secretory granules which have a highly constrained volume; (2) transformation of such immature granules into mature secretory granules, a process often associated with a reduction in the maturing granule's volume, as well as changes in the appearance of its content and (3) fusion of secretory granules of the smallest size, termed 'unit granules', forming granules whose volumes are multiples of the unit granule's volume. Mutations which perturb this process can cause significant pathology. For example, Chediak-Higashi syndrome / lysosomal trafficking regulator (CHS)/(Lyst) mutations result in giant secretory granules in a number of cell types in human beings with the Chediak-Higashi syndrome and in 'beige' (Lyst(bg)/Lyst(bg)) mice. Analysis of the secretory granules of mast cells and pancreatic acinar cells in Lyst-deficient beige mice suggests that beige mouse secretory granules retain the ability to fuse randomly with other secretory granules no matter what the size of the fusion partners. By contrast, in normal mice, the pattern of granule-granule fusion occurs exclusively by the addition of unit granules, either to each other or to larger granules. The normal pattern of fusion is termed unit addition and the fusion evident in cells with CHS/Lyst mutations is called random addition. The proposed model of secretory granule formation has several implications. For example, in neurosecretory cells, the secretion of small amounts of cargo in granules constrained to a very narrow size increases the precision of the information conveyed by secretion. By contrast, in pancreatic acinar cells and mast cells, large granules composed of multiple unit granules permit the cells to store large amounts of material without requiring the amount of membrane necessary to package the same amount of cargo into small granules. In addition, the formation of mature secretory granules that are multimers of unit granules provides a mechanism for mixing in large granules the contents of unit granules which differ in their content of cargo.

    View details for DOI 10.1111/j.1582-4934.2010.01071.x

    View details for Web of Science ID 000281141900003

    View details for PubMedID 20406331

  • Distinguishing Mast Cell and Granulocyte Differentiation at the Single-Cell Level CELL STEM CELL Franco, C. B., Chen, C., Drukker, M., Weissman, I. L., Galli, S. J. 2010; 6 (4): 361-368

    Abstract

    The lineage restriction of prospectively isolated hematopoietic progenitors has been traditionally assessed by bulk in vitro culture and transplantation of large number of cells in vivo. These methods, however, cannot distinguish between homogenous multipotent or heterogeneous lineage-restricted populations. Using clonal assays of 1 or 5 cells in vitro, single-cell quantitative gene expression analyses, and transplantation of mice with low numbers of cells, we show that a common myeloid progenitor (CMP) is Sca-1(lo)lin(-)c-Kit(+)CD27(+)Flk-2(-) (SL-CMP; Sca-1(lo) CMP) and a granulocyte/macrophage progenitor (GMP) is Sca-1(lo)lin(-)c-Kit(+)CD27(+)Flk-2(+)CD150(-/lo) (SL-GMP; Sca-1(lo) GMP). We found that mast cell progenitor potential is present in the SL-CMP fraction, but not in the more differentiated SL-GMP population, and is more closely related to megakaryocyte/erythrocyte specification. Our data provide criteria for the prospective isolation of SL-CMP and SL-GMP and support the conclusion that mast cells are specified during hematopoiesis earlier than and independently from granulocytes.

    View details for DOI 10.1016/j.stem.2010.02.013

    View details for Web of Science ID 000276823300014

    View details for PubMedID 20362540

  • The role of recipient mast cells in acute and chronic cardiac allograft rejection in C57BL/6-KitW-sh/W-sh mice JOURNAL OF HEART AND LUNG TRANSPLANTATION Itoh, S., Nakae, S., Velotta, J. B., Kosuge, H., Connolly, A., Tsai, M., Adachi, H., Galli, S. J., Robbins, R. C., Fischbein, M. P. 2010; 29 (4): 401-409

    Abstract

    Mast cells are hypothesized to promote rejection and adverse remodeling in cardiac allografts. In contrast, it has been reported that mast cells may enhance cardiac allograft survival in rats. We used C57BL/6-Kit(W-sh/W-sh) mast cell-deficient and corresponding wild-type mice to investigate possible contributions of recipient mast cells to acute or chronic cardiac allograft rejection.FVB (H-2(q); acute rejection), or C-H-2(bm12)KhEg (H-2(bm12); chronic rejection) donor hearts were heterotopically transplanted into C57BL/6-Kit(W-sh/W-sh) (H-2(b)) or C57BL/6-Kit(+/+) (H-2(b)) mice. The degree of acute rejection was assessed at 5 days and chronic rejection, at 52 days.In the acute rejection model, donor heart vascular cell adhesion molecule-1 (VCAM-1) expression was significantly lower in C57BL/6-Kit(W-sh/W-sh) than in wild-type recipients; however, acute rejection scores, graft survival, inflammatory cells, or cytokine expression did not differ significantly. In the chronic rejection model, the number of mast cells/mm(2) of allograft tissue was significantly increased 52 days after transplantation in allografts transplanted into C57BL/6-Kit(+/+) but not C57BL/6-Kit(W-sh/W-sh) mice; however, no substantial differences were noted in graft coronary artery disease, graft inflammatory cells, or levels of graft tissue expression of cytokines or adhesion molecules.Cardiac allografts undergoing chronic rejection in wild-type C57BL/6-Kit(+/+) mice exhibit increased numbers of mast cells, but acute or chronic cardiac allograft rejection can develop in C57BL/6-Kit(W-sh/W-sh) mice even though these recipients virtually lack mast cells. These findings indicate that recipient mast cells are not required for acute or chronic cardiac allograft rejection in the models examined.

    View details for DOI 10.1016/j.healun.2009.08.019

    View details for Web of Science ID 000276915100003

    View details for PubMedID 19818646

  • Evidence that vitamin D-3 promotes mast cell-dependent reduction of chronic UVB-nduced skin pathology in mice JOURNAL OF EXPERIMENTAL MEDICINE Biggs, L., Yu, C., Fedoric, B., Lopez, A. F., Galli, S. J., Grimbaldeston, M. A. 2010; 207 (3): 455-463

    Abstract

    Mast cell production of interleukin-10 (IL-10) can limit the skin pathology induced by chronic low-dose ultraviolet (UV)-B irradiation. Although the mechanism that promotes mast cell IL-10 production in this setting is unknown, exposure of the skin to UVB irradiation induces increased production of the immune modifying agent 1alpha,25-dihydroxyvitamin D(3) (1alpha,25[OH](2)D(3)). We now show that 1alpha,25(OH)(2)D(3) can up-regulate IL-10 mRNA expression and induce IL-10 secretion in mouse mast cells in vitro. To investigate the roles of 1alpha,25(OH)(2)D(3) and mast cell vitamin D receptor (VDR) expression in chronically UVB-irradiated skin in vivo, we engrafted the skin of genetically mast cell-deficient WBB6F(1)-Kit(W/W-v) mice with bone marrow-derived cultured mast cells derived from C57BL/6 wild-type or VDR(-/-) mice. Optimal mast cell-dependent suppression of the inflammation, local production of proinflammatory cytokines, epidermal hyperplasia, and epidermal ulceration associated with chronic UVB irradiation of the skin in Kit(W/W-v) mice required expression of VDR by the adoptively transferred mast cells. Our findings suggest that 1alpha,25(OH)(2)D(3)/VDR-dependent induction of IL-10 production by cutaneous mast cells can contribute to the mast cell's ability to suppress inflammation and skin pathology at sites of chronic UVB irradiation.

    View details for DOI 10.1084/jem.20091725

    View details for Web of Science ID 000275593900003

    View details for PubMedID 20194632

  • Mast Cell-Derived TNF Can Exacerbate Mortality during Severe Bacterial Infections in C57BL/6-KitW-sh/W-sh Mice AMERICAN JOURNAL OF PATHOLOGY Piliponsky, A. M., Chen, C., Grimbaldeston, M. A., Burns-Guydish, S. M., Hardy, J., Kalesnikoff, J., Contag, C. H., Tsai, M., Galli, S. J. 2010; 176 (2): 926-938

    Abstract

    We used mast cell-engrafted genetically mast cell-deficient C57BL/6-Kit(W-sh/W-sh) mice to investigate the roles of mast cells and mast cell-derived tumor necrosis factor in two models of severe bacterial infection. In these mice, we confirmed findings derived from studies of mast cell-deficient WBB6F(1)-Kit(W/W-v) mice indicating that mast cells can promote survival in cecal ligation and puncture (CLP) of moderate severity. However, we found that the beneficial role of mast cells in this setting can occur independently of mast cell-derived tumor necrosis factor. By contrast, using mast cell-engrafted C57BL/6-Kit(W-sh/W-sh) mice, we found that mast cell-derived tumor necrosis factor can increase mortality during severe CLP and can also enhance bacterial growth and hasten death after intraperitoneal inoculation of Salmonella typhimurium. In WBB6F(1)-Kit(W-sh/W-sh) mice, mast cells enhanced survival during moderately severe CLP but did not significantly change the survival observed in severe CLP. Our findings in three types of genetically mast cell-deficient mice thus support the hypothesis that, depending on the circumstances (including mouse strain background, the nature of the mutation resulting in a mast cell deficiency, and type and severity of infection), mast cells can have either no detectable effect or opposite effects on survival during bacterial infections, eg, promoting survival during moderately severe CLP associated with low mortality but, in C57BL/6-Kit(W-sh/W-sh) mice, increasing mortality during severe CLP or infection with S. typhimurium.

    View details for DOI 10.2353/ajpath.2010.090342

    View details for Web of Science ID 000274111400040

    View details for PubMedID 20035049

  • Anaphylaxis: mechanisms of mast cell activation. Chemical immunology and allergy Kalesnikoff, J., Galli, S. J. 2010; 95: 45-66

    Abstract

    Anaphylaxis is a severe systemic allergic response that is rapid in onset and potentially lethal, and that typically is induced by an otherwise innocuous substance. In IgE-dependent and other examples of anaphylaxis, tissue mast cells and circulating basophilic granulocytes (basophils) are thought to represent major (if not the major) sources of the biologically active mediators that contribute to the pathology and, in unfortunate individuals, fatal outcome, of anaphylaxis. In this chapter, we will describe the mechanisms of mast cell (and basophil) activation in anaphylaxis, with a focus on IgE-dependent activation, which is thought to be responsible for most examples of antigen-induced anaphylaxis in humans. We will also discuss the use of mouse models to investigate the mechanisms that can contribute to anaphylaxis in that species in vivo, and the relevance of such mouse studies to human anaphylaxis.

    View details for DOI 10.1159/000315937

    View details for PubMedID 20519881

  • Pillars article: fate of bone marrow-derived cultured mast cells after intracutaneous, intraperitoneal, and intravenous transfer into genetically mast cell-deficient w/wv mice. Evidence that cultured mast cells can give rise to both connective tissue type and mucosal mast cells. Journal of immunology Nakano, T., Sonoda, T., Hayashi, C., Yamatodani, A., Kanayama, Y., Yamamura, T., Asai, H., Yonezawa, T., Kitamura, Y., Galli, S. J. 2009; 183 (11): 6863-6881

    View details for PubMedID 19923473

  • Rabaptin-5 regulates receptor expression and functional activation in mast cells BLOOD Rios, E. J., Piliponsky, A. M., Ra, C., Kalesnikoff, J., Galli, S. J. 2008; 112 (10): 4148-4157

    Abstract

    Rab5 is a small GTPase that regulates early endocytic events and is activated by RabGEF1/Rabex-5. Rabaptin-5, a Rab5 interacting protein, was identified as a protein critical for potentiating RabGEF1/Rabex-5's activation of Rab5. Using Rabaptin-5 shRNA knockdown, we show that Rabaptin-5 is dispensable for Rab5-dependent processes in intact mast cells, including high affinity IgE receptor (FcepsilonRI) internalization and endosome fusion. However, Rabaptin-5 deficiency markedly diminished expression of FcepsilonRI and beta1 integrin on the mast cell surface by diminishing receptor surface stability. This in turn reduced the ability of mast cells to bind IgE and significantly diminished both mast cell sensitivity to antigen (Ag)-induced mediator release and Ag-induced mast cell adhesion and migration. These findings show that, although dispensable for canonical Rab5 processes in mast cells, Rabaptin-5 importantly contributes to mast cell IgE-dependent immunologic function by enhancing mast cell receptor surface stability.

    View details for DOI 10.1182/blood-2008-04-152660

    View details for Web of Science ID 000260691300035

    View details for PubMedID 18698003

  • New developments in mast cell biology NATURE IMMUNOLOGY Kalesnikoff, J., Galli, S. J. 2008; 9 (11): 1215-1223

    Abstract

    Mast cells can function as effector and immunoregulatory cells in immunoglobulin E-associated allergic disorders, as well as in certain innate and adaptive immune responses. This review focuses on exciting new developments in the field of mast cell biology published in the past year. We highlight advances in the understanding of FcvarepsilonRI-mediated signaling and mast cell-activation events, as well as in the use of genetic models to study mast cell function in vivo. Finally, we discuss newly identified functions for mast cells or individual mast cell products, such as proteases and interleukin 10, in host defense, cardiovascular disease and tumor biology and in settings in which mast cells have anti-inflammatory or immunosuppressive functions.

    View details for DOI 10.1038/ni.f.216

    View details for Web of Science ID 000260248600008

    View details for PubMedID 18936782

  • IL-3 is required for increases in blood basophils in nematode infection in mice and can enhance IgE-dependent IL-4 production by basophils in vitro LABORATORY INVESTIGATION Lantz, C. S., Min, B., Tsai, M., Chatterjea, D., Dranoff, G., Galli, S. J. 2008; 88 (11): 1134-1142

    Abstract

    Basophils represent potential effector and immunoregulatory cells, as well as a potential source of IL-4, during the immune response elicited by infection with the nematode Nippostrongylus brasiliensis (N.b.), and in other settings. However, the factors which regulate the numbers of blood basophils in mice, or the ability of these cells to produce IL-4, are not fully understood. We found that infection of mice with the nematodes N.b. or Strongyloides venezuelensis (S.v.) induced substantial increases in the numbers of blood basophils (to as high as 18% of circulating blood leukocytes). Experiments in IL-3-/- vs IL-3+/+ mice, and in IL-3-treated IL-3-/- mice, showed that essentially all of the increases in blood or bone marrow basophils during N.b. or S.v. infection were IL-3 dependent. Many of the blood, bone marrow or liver-derived basophils from IL-3-/- or IL-3+/+ mice expressed intracellular IL-4 upon stimulation with anti-IgE in vitro. However, after incubation of the cells with exogenous IgE in vitro, blood- or liver-derived basophils from IL-3+/+ mice exhibited higher levels of intracellular IL-4 after stimulation with anti-IgE than did basophils derived from IL-3-/- mice. Thus, IL-3 is a major regulator of the marked increases in blood basophil levels observed during infection of mice with N.b. or S.v. and also can enhance levels of intracellular IL-4 upon activation of basophils with anti-IgE in vitro.

    View details for DOI 10.1038/labinvest.2008.88

    View details for Web of Science ID 000260427600001

    View details for PubMedID 18975389

  • Mast cell-expressed orphan receptor CCRL2 binds chemerin and is required for optimal induction of IgE-mediated passive cutaneous anaphylaxis JOURNAL OF EXPERIMENTAL MEDICINE Zabel, B. A., Nakae, S., Zuniga, L., Kim, J., Ohyama, T., Alt, C., Pan, J., Suto, H., Soler, D., Allen, S. J., Handel, T. M., Song, C. H., Galli, S. J., Butcher, E. C. 2008; 205 (10): 2207-2220

    Abstract

    Mast cells contribute importantly to both protective and pathological IgE-dependent immune responses. We show that the mast cell-expressed orphan serpentine receptor mCCRL2 is not required for expression of IgE-mediated mast cell-dependent passive cutaneous anaphylaxis but can enhance the tissue swelling and leukocyte infiltrates associated with such reactions in mice. We further identify chemerin as a natural nonsignaling protein ligand for both human and mouse CCRL2. In contrast to other "silent" or professional chemokine interreceptors, chemerin binding does not trigger ligand internalization. Rather, CCRL2 is able to bind the chemoattractant and increase local concentrations of bioactive chemerin, thus providing a link between CCRL2 expression and inflammation via the cell-signaling chemerin receptor CMKLR1.

    View details for DOI 10.1084/jem.20080300

    View details for Web of Science ID 000259656700005

    View details for PubMedID 18794339

  • The development of allergic inflammation NATURE Galli, S. J., Tsai, M., Piliponsky, A. M. 2008; 454 (7203): 445-454

    Abstract

    Allergic disorders, such as anaphylaxis, hay fever, eczema and asthma, now afflict roughly 25% of people in the developed world. In allergic subjects, persistent or repetitive exposure to allergens, which typically are intrinsically innocuous substances common in the environment, results in chronic allergic inflammation. This in turn produces long-term changes in the structure of the affected organs and substantial abnormalities in their function. It is therefore important to understand the characteristics and consequences of acute and chronic allergic inflammation, and in particular to explore how mast cells can contribute to several features of this maladaptive pattern of immunological reactivity.

    View details for DOI 10.1038/nature07204

    View details for Web of Science ID 000257860300039

    View details for PubMedID 18650915

  • Immunomodulatory mast cells: negative, as well as positive, regulators of immunity NATURE REVIEWS IMMUNOLOGY Galli, S. J., Grimbaldeston, M., Tsai, M. 2008; 8 (6): 478-U14

    Abstract

    Mast cells can promote inflammation and other tissue changes in IgE-associated allergic disorders, as well as in certain innate and adaptive immune responses that are thought to be independent of IgE. However, mast cells can also have anti-inflammatory and immunosuppressive functions. Here, we review the evidence that mast cells can have negative, as well as positive, immunomodulatory roles in vivo, and we propose that mast cells can both enhance and later suppress certain features of an immune response.

    View details for DOI 10.1038/nri2327

    View details for Web of Science ID 000256105700018

    View details for PubMedID 18483499

  • Neurotensin increases mortality and mast cells reduce neurotensin levels in a mouse model of sepsis NATURE MEDICINE Piliponsky, A. M., Chen, C., Nishimura, T., Metz, M., Rios, E. J., Dobner, P. R., Wada, E., Wada, K., Zacharias, S., Mohanasundaram, U. M., Faix, J. D., Abrink, M., Pejler, G., Pearl, R. G., Tsai, M., Galli, S. J. 2008; 14 (4): 392-398

    Abstract

    Sepsis is a complex, incompletely understood and often fatal disorder, typically accompanied by hypotension, that is considered to represent a dysregulated host response to infection. Neurotensin (NT) is a 13-amino-acid peptide that, among its multiple effects, induces hypotension. We find that intraperitoneal and plasma concentrations of NT are increased in mice after severe cecal ligation and puncture (CLP), a model of sepsis, and that mice treated with a pharmacological antagonist of NT, or NT-deficient mice, show reduced mortality during severe CLP. In mice, mast cells can degrade NT and reduce NT-induced hypotension and CLP-associated mortality, and optimal expression of these effects requires mast cell expression of neurotensin receptor 1 and neurolysin. These findings show that NT contributes to sepsis-related mortality in mice during severe CLP and that mast cells can lower NT concentrations, and suggest that mast cell-dependent reduction in NT levels contributes to the ability of mast cells to enhance survival after CLP.

    View details for DOI 10.1038/nm1738

    View details for Web of Science ID 000254674100025

    View details for PubMedID 18376408

  • Basophils are back! IMMUNITY Galli, S. J., Franco, C. B. 2008; 28 (4): 495-497

    Abstract

    In this issue of Immunity, Tsujimura et al. (2008) report that the release of platelet-activating factor by basophils stimulated with immunoglobulin G1 (IgG1)-antigen immune complexes contributes substantially to the expression of an IgG1-dependent alternative pathway of systemic anaphylaxis in mice.

    View details for DOI 10.1016/j.immuni.2008.03.010

    View details for Web of Science ID 000254939400010

    View details for PubMedID 18400194

  • Mast cells: Versatile regulators of inflammation, tissue remodeling, host defense and homeostasis JOURNAL OF DERMATOLOGICAL SCIENCE Galli, S. J., Tsai, M. 2008; 49 (1): 7-19

    Abstract

    The possible roles of mast cells in heath and disease have been a topic of interest for over 125 years. Many adaptive or pathological processes affecting the skin or other anatomical sites have been associated with morphological evidence of mast cell activation, and/or with changes in mast cell numbers or phenotype. Such observations, taken together with the known functions of the diverse mediators, cytokines and growth factors which can be secreted by mast cells, have suggested many potential functions for mast cells in health and disease. Definitively identifying the importance of mast cells in biological responses in humans is difficult. However, mutant mice which are profoundly mast cell-deficient, especially those which can undergo engraftment with wild-type or genetically altered mast cells, provide an opportunity to investigate the importance of mast cells, and specific mast cell functions or products, in various adaptive or pathological responses in mice. Such work has shown that mast cells can significantly influence multiple features of inflammatory or immune responses, through diverse effects that can either promote or, surprisingly, suppress, aspects of these responses. Through such functions, mast cells can significantly influence inflammation, tissue remodeling, host defense and homeostasis.

    View details for DOI 10.1016/j.jdermsci.2007.09.009

    View details for Web of Science ID 000252523500003

    View details for PubMedID 18024086

  • IL-33 induces IL-13 production by mouse mast cells independently of IgE-Fc epsilon RI signals JOURNAL OF LEUKOCYTE BIOLOGY Ho, L. H., Ohno, T., Oboki, K., Kajiwara, N., Suto, H., Iikura, M., Okayama, Y., Akira, S., Saito, H., Galli, S. J., Nakae, S. 2007; 82 (6): 1481-1490

    Abstract

    The IL-1-related molecules, IL-1 and IL-18, can promote Th2 cytokine production by IgE/antigen-FcepsilonRI-stimulated mouse mast cells. Another IL-1-related molecule, IL-33, was identified recently as a ligand for T1/ST2. Although mouse mast cells constitutively express ST2, the effects of IL-33 on mast cell function are poorly understood. We found that IL-33, but not IL-1beta or IL-18, induced IL-13 and IL-6 production by mouse bone marrow-derived, cultured mast cells (BMCMCs) independently of IgE. In BMCMCs incubated with the potently cytokinergic SPE-7 IgE without specific antigen, IL-33, IL-1beta, and IL-18 each promoted IL-13 and IL-6 production, but the effects of IL-33 were more potent than those of IL-1beta or IL-18. IL-33 promoted cytokine production via a MyD88-dependent but Toll/IL-1R domain-containing adaptor-inducing IFN-beta-independent pathway. By contrast, IL-33 neither induced nor enhanced mast cell degranulation. At 200 ng/ml, IL-33 prolonged mast cell survival in the absence of IgE and impaired survival in the presence of SPE-7 IgE, whereas at 100 ng/ml, IL-33 had no effect on mast cell survival in the absence of IgE and reduced mast cell survival in the presence of IgE. These observations suggest potential roles for IL-33 in mast cell- and Th2 cytokine-associated immune responses and disorders.

    View details for DOI 10.1189/jlb.0407200

    View details for Web of Science ID 000251243800016

    View details for PubMedID 17881510

  • TIM-1 and TIM-3 enhancement of Th2 cytokine poduction by mast cells BLOOD Nakae, S., Iikura, M., Suto, H., Akiba, H., Umetsu, D. T., DeKruyff, R. H., Saito, H., Galli, S. J. 2007; 110 (7): 2565-2568

    Abstract

    Members of the T-cell immunoglobulin- and mucin-domain-containing molecule (TIM) family have roles in T-cell-mediated immune responses. TIM-1 and TIM-2 are predominantly expressed on T helper type 2 (Th2) cells, whereas TIM-3 is preferentially expressed on Th1 and Th17 cells. We found that TIM-1 and TIM-3, but neither TIM-2 nor TIM-4, were constitutively expressed on mouse peritoneal mast cells and bone marrow-derived cultured mast cells (BMCMCs). After IgE + Ag stimulation, TIM-1 expression was down-regulated on BMCMCs, whereas TIM-3 expression was up-regulated. We also found that recombinant mouse TIM-4 (rmTIM-4), which is a ligand for TIM-1, as well as an anti-TIM-3 polyclonal Ab, can promote interleukin-4 (IL-4), IL-6, and IL-13 production without enhancing degranulation in BMCMCs stimulated with IgE + Ag. Moreover, the anti-TIM-3 Ab, but neither anti-TIM-1 Ab nor rmTIM-4, suppressed mast-cell apoptosis. These observations suggest that TIM-1 and TIM-3 may be able to influence T-cell-mediated immune responses in part through effects on mast cells.

    View details for DOI 10.1182/blood-2006-11-058800

    View details for Web of Science ID 000249800900056

    View details for PubMedID 17620455

  • IL-33 can promote survival, adhesion and cytokine production in human mast cells LABORATORY INVESTIGATION Iikura, M., Suto, H., Kajiwara, N., Oboki, K., Ohno, T., Saito, H., Galli, S. J., Nakae, S. 2007; 87 (10): 971-978

    Abstract

    IL-33 is a recently identified member of the IL-1 family of molecules, which also includes IL-1 and IL-18. IL-33 binds to the receptor, T1/ST2/IL-1R4, and can promote cytokine secretion by Th2 cells and NF-kappaB phosphorylation in mouse mast cells. However, the effects of these molecules, especially IL-33, in human mast cells are poorly understood. Expression of the receptors for IL-1 family molecules, specifically, IL-1R1, IL-18R and T1/ST2, was detectable intracellularly in human umbilical cord blood-derived mast cells (HUCBMCs) by flow cytometry, but was scarcely detectable on the cells' surface. However, IL-1beta, IL-18 or IL-33 induced phosphorylation of Erk, p38 and JNK in na´ve HUCBMCs, and IL-33 or IL-1beta, but not IL-18, enhanced the survival of naive HUCBMCs and promoted their adhesion to fibronectin. IL-33 or IL-1beta also induced IL-8 and IL-13 production in na´ve HUCBMCs, and enhanced production of these cytokines in IgE/anti-IgE-stimulated HUCBMCs, without enhancing secretion of either PGD(2) or histamine. Moreover, IL-33-mediated IL-8 production by HUCBMCs was markedly reduced by the p38 MAPK inhibitor, SB203580. In contrast to findings with mouse mast cells, IL-18 neither induced nor enhanced secretion of the mediators PGD(2) or histamine by HUCBMCs. Our findings identify previously unknown functions of IL-33 in human mast cells. One of these is that IL-33, like IL-1beta, can induce cytokine production in human mast cells even in the absence of stimuli of FcepsilonRI aggregation. Our findings thus support the hypothesis that IL-33 may enhance mast cell function in allergic disorders and other settings, either in the presence or absence of co-stimulation of mast cells via IgE/antigen-FcepsilonRI signals.

    View details for DOI 10.1038/labinvest.3700663

    View details for Web of Science ID 000249557400002

    View details for PubMedID 17700564

  • Mast cell-derived interleukin 10 limits skin pathology in contact dermatitis and chronic irradiation with ultraviolet B NATURE IMMUNOLOGY Grimbaldeston, M. A., Nakae, S., Kalesnikoff, J., Tsai, M., Galli, S. J. 2007; 8 (10): 1095-1104

    Abstract

    Allergic contact dermatitis, such as in response to poison ivy or poison oak, and chronic low-dose ultraviolet B irradiation can damage the skin. Mast cells produce proinflammatory mediators that are thought to exacerbate these prevalent acquired immune or innate responses. Here we found that, unexpectedly, mast cells substantially limited the pathology associated with these responses, including infiltrates of leukocytes, epidermal hyperplasia and epidermal necrosis. Production of interleukin 10 by mast cells contributed to the anti-inflammatory or immunosuppressive effects of mast cells in these conditions. Our findings identify a previously unrecognized function for mast cells and mast cell-derived interleukin 10 in limiting leukocyte infiltration, inflammation and tissue damage associated with immunological or innate responses that can injure the skin.

    View details for DOI 10.1038/ni1503

    View details for Web of Science ID 000249691400024

    View details for PubMedID 17767162

  • Mast cell-derived TNF contributes to airway hyperreactivity, inflammation, and T(H)2 cytokine production in an asthma model in mice JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY Nakae, S., Ho, L. H., Yu, M., Monteforte, R., Iikura, M., Suto, H., Galli, S. J. 2007; 120 (1): 48-55

    Abstract

    Mast cells, IgE, and TNF, which have been implicated in human atopic asthma, contribute significantly to the allergic airway inflammation induced by ovalbumin (OVA) challenge in mice sensitized with OVA without alum. However, it is not clear to what extent mast cells represent a significant source of TNF in this mouse model.We investigated the importance of mast cell-derived TNF in a mast cell-dependent model of OVA-induced airway hyperreactivity (AHR) and allergic airway inflammation.Features of this model of airway inflammation were analyzed in C57BL/6J-wild-type mice, mast cell-deficient C57BL/6J-Kit(W-sh)(/W-sh) mice, and C57BL/6J Kit(W-sh/W-sh) mice that had been systemically engrafted with bone marrow-derived cultured mast cells from C57BL/6J-wild-type or C57BL/6J-TNF(-/-) mice.Ovalbumin-induced AHR and airway inflammation were significantly reduced in mast cell-deficient Kit(W-sh/W-sh) mice versus wild-type mice. By contrast, Kit(W-sh/W-sh) mice that had been engrafted with wild-type but not with TNF(-/-) bone marrow-derived cultured mast cells exhibited responses very similar to those observed in wild-type mice. Mast cells and mast cell-derived TNF were not required for induction of OVA-specific memory T cells in the sensitization phase, but significantly enhanced lymphocyte recruitment and T(H)2 cytokine production in the challenge phase.Mast cell-derived TNF contributes significantly to the pathogenesis of mast cell-dependent and IgE-dependent, OVA-induced allergic inflammation and AHR in mice, perhaps in part by enhancing lymphocyte recruitment and T(H)2 cytokine production.Our findings in mice support the hypothesis that mast cell-derived TNF can promote allergic inflammation and AHR in asthma.

    View details for DOI 10.1016/j.jaci.2007.02.046

    View details for Web of Science ID 000248066400007

    View details for PubMedID 17482668

  • Roles of RabGEF1/Rabex-5 domains in regulating Fc epsilon RI surface expression and Fc epsilon RI-dependent responses in mast cells BLOOD Kalesnikoff, J., Rios, E. J., Chen, C., Barbieri, M. A., Tsai, M., Tam, S., Galli, S. J. 2007; 109 (12): 5308-5317

    Abstract

    RabGEF1/Rabex-5, a guanine nucleotide exchange factor (GEF) for the endocytic pathway regulator, Rab5, contains a Vps9 domain, an A20-like zinc finger (ZnF) domain, and a coiled coil domain. To investigate the importance of these domains in regulating receptor internalization and cell activation, we lentivirally delivered RabGEF1 mutants into RabGEF1-deficient (-/-) mast cells and examined Fc epsilon RI-dependent responses. Wild-type RabGEF1 expression corrected phenotypic abnormalities in -/- mast cells, including decreased basal Fc epsilon RI expression, slowed Fc epsilon RI internalization, elevated IgE + Ag-induced degranulation and IL-6 production, and the decreased ability of -/- cytosol to support endosome fusion. We showed that RabGEF1's ZnF domain has ubiquitin ligase activity. Moreover, the coiled coil domain of RabGEF1 is required for Rabaptin-5 binding and for maintaining basal levels of Rabaptin-5 and surface Fc epsilon RI. However, mutants lacking either of these domains normalized phenotypic abnormalities in IgE + antigen-activated -/- mast cells. By contrast, correction of these -/- phenotypes required a functional Vps9 domain. Thus, Fc epsilon RI-mediated mast cell functional activation is dependent on RabGEF1's GEF activity.

    View details for DOI 10.1182/blood-2007-01-067363

    View details for Web of Science ID 000247360200045

    View details for PubMedID 17341663

  • Mast cells in the promotion and limitation of chronic inflammation IMMUNOLOGICAL REVIEWS Metz, M., Grimbaldeston, M. A., Nakae, S., Piliponsky, A. M., Tsai, M., Galli, S. J. 2007; 217: 304-328

    Abstract

    Observations of increased numbers of mast cells at sites of chronic inflammation have been reported for over a hundred years. Light and electron microscopic evidence of mast cell activation at such sites, taken together with the known functions of the diverse mediators, cytokines, and growth factors that can be secreted by appropriately activated mast cells, have suggested a wide range of possible functions for mast cells in promoting (or suppressing) many features of chronic inflammation. Similarly, these and other lines of evidence have implicated mast cells in a variety of adaptive or pathological responses that are associated with persistent inflammation at the affected sites. Definitively characterizing the importance of mast cells in chronic inflammation in humans is difficult. However, mice that genetically lack mast cells, especially those which can undergo engraftment with wildtype or genetically altered mast cells, provide a means to investigate the importance of mast cells and specific mast cell functions or products in diverse models of chronic inflammation. Such work has confirmed that mast cells can significantly influence multiple features of chronic inflammatory responses, through diverse effects that can either promote or, perhaps more surprisingly, suppress aspects of these responses.

    View details for Web of Science ID 000246317100022

    View details for PubMedID 17498068

  • Mast cell-derived TNF can promote Th17 cell-dependent neutrophil recruitment in ovalbumin-challenged OTII mice BLOOD Nakae, S., Suto, H., Berry, G. J., Galli, S. J. 2007; 109 (9): 3640-3648

    Abstract

    Both mast cells and IL-17 can contribute to host defense and pathology in part by orchestrating neutrophil recruitment, but the possible role of mast cells in IL-17-induced inflammation remains to be defined. We found that mast cells and IL-17, but neither IFN-gamma nor FcRgamma signaling, contributed significantly to the antigen (Ag)-dependent airway neutrophilia elicited in ovalbumin-specific T-cell receptor (TCR)-expressing C57BL/6-OTII mice, and that IFN-gamma significantly suppressed IL-17-dependent airway neutrophilia in this setting. IL-18, IL-1beta, and TNF each contributed significantly to the development of Ag- and T helper 17 (Th17 cell)-mediated airway neutrophilia. Moreover, IL-17 enhanced mast cell TNF production in vitro, and mast cell-associated TNF contributed significantly to Ag- and Th17 cell-mediated airway neutrophilia in vivo. By contrast, we detected no significant role for the candidate mediators histamine, PGD(2), LTB(4), CXCL10, or IL-16, each of which can be produced by mast cells and other cell types, in the neutrophil infiltration elicited in this model. These findings establish that mast cells and mast cell-derived TNF can significantly enhance, by FcRgamma-independent mechanisms, the Ag- and Th17 cell-dependent development of a neutrophil-rich inflammatory response at a site of Ag challenge.

    View details for DOI 10.1182/blood-2006-09-046128

    View details for Web of Science ID 000246091400014

    View details for PubMedID 17197430

  • TNF can contribute to multiple features of ovalbumin-induced allergic inflammation of the airways in mice JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY Nakae, S., Lunderius, C., Ho, L. H., Schafer, B., Tsai, M., Galli, S. J. 2007; 119 (3): 680-686

    Abstract

    TNF is thought to contribute to airway hyperreactivity (AHR) and airway inflammation in asthma. However, studies with TNF-deficient or TNF receptor-deficient mice have not produced a clear picture of the role of TNF in the AHR associated with allergic inflammation in the mouse.We used a genetic approach to investigate the contributions of TNF to antigen-induced AHR and airway inflammation in mice on the C57BL/6 background.We analyzed features of airway allergic inflammation, including antigen-induced AHR, in C57BL/6 wild-type and TNF(-/-) mice, using 2 different methods for sensitizing the mice to ovalbumin (OVA).In mice sensitized to OVA administered with the adjuvant aluminum hydroxide (alum), which develop IgE-independent and mast cell-independent allergic inflammation and AHR, we found no significant differences in OVA-induced AHR in C57BL/6-TNF(-/-) versus wild-type mice. By contrast, in mice sensitized to OVA without alum, which develop allergic inflammation that is significantly mast cell-dependent, C57BL/6-TNF(-/-) mice exhibited significant reductions versus wild-type mice in OVA-induced AHR to methacholine; numbers of lymphocytes, neutrophils, and eosinophils in bronchoalveolar lavage fluid; levels of myeloperoxidase, eosinophil peroxidase, and the cytokines IL-4, IL-5, and IL-17 in lung tissue; and histologic evidence of pulmonary inflammation.In pulmonary allergic inflammation induced in mice immunized with OVA without alum, TNF significantly contributes to several features of the response, including antigen-induced inflammation and AHR.Our findings in mice support the hypothesis that TNF can promote the allergic inflammation and AHR associated with asthma.

    View details for DOI 10.1016/j.jaci.2006.11.701

    View details for Web of Science ID 000244925000022

    View details for PubMedID 17336618

  • Effector and potential immunoregulatory roles of mast cells in IgE-associated acquired immune responses CURRENT OPINION IN IMMUNOLOGY Grimbaldeston, M. A., Metz, M., Yu, M., Tsai, M., Galli, S. J. 2006; 18 (6): 751-760

    Abstract

    Mast cells are best known as critical effector cells in anaphylaxis and other examples of IgE-associated immediate hypersensitivity reactions. However, mast cells also can contribute to the development of the late-phase responses that occur in some sensitized subjects hours after initial exposure to specific antigen, and can promote many of the features of chronic allergic inflammation, including tissue remodeling and functional changes in the affected organs. In addition to such effector cell functions in IgE-associated immune responses, recent evidence indicates that mast cells can importantly influence the sensitization phase of at least some acquired immune responses, and can contribute to the pathology of autoimmune disorders and to the expression of peripheral tolerance.

    View details for DOI 10.1016/j.coi.2006.09.011

    View details for Web of Science ID 000242036900018

    View details for PubMedID 17011762

  • Mast cell-derived tumor necrosis factor can promote nerve fiber elongation in the skin during contact hypersensitivity in mice AMERICAN JOURNAL OF PATHOLOGY Kakurai, M., Monteforte, R., Suto, H., Tsai, M., Nakae, S., Galli, S. J. 2006; 169 (5): 1713-1721

    Abstract

    In humans, lesions of contact eczema or atopic dermatitis can exhibit increases in epidermal nerves, but the mechanism resulting in such nerve elongation are not fully understood. We found that contact hypersensitivity reactions to oxazolone in mice were associated with significant increases in the length of nerves in the epidermis and dermis. Using genetically mast cell-deficient c-kit mutant mice selectively repaired of their dermal mast cell deficiency with either wild-type or tumor necrosis factor (TNF)-deficient mast cells, we found that mast cells, and mast cell-derived TNF, significantly contributed to the elongation of epidermal and dermal PGP 9.5+ nerves and dermal CGRP+ nerves, as well as to the inflammation observed at sites of contact hypersensitivity in response to oxazolone. Moreover, the percentage of mast cells in close proximity to dermal PGP 9.5+ nerve fibers was significantly higher in wild-type mice and in c-kit mutant mice repaired of their dermal mast cell deficiency by the adoptive transfer of wild-type mast cells than in TNF-deficient mice or in TNF-/- mast cell-engrafted c-kit mutant mice. These observations show that mast cells, and mast cell-derived TNF, can promote the elongation of cutaneous nerve fibers during contact hypersensitivity in the mouse.

    View details for DOI 10.2353/ajpath.2006.060602

    View details for Web of Science ID 000241603700019

    View details for PubMedID 17071594

  • Mast cells can enhance resistance to snake and honeybee venoms SCIENCE Metz, M., Piliponsky, A. M., Chen, C., Lammel, V., Abrink, M., Pejler, G., Tsai, M., Galli, S. J. 2006; 313 (5786): 526-530

    Abstract

    Snake or honeybee envenomation can cause substantial morbidity and mortality, and it has been proposed that the activation of mast cells by snake or insect venoms can contribute to these effects. We show, in contrast, that mast cells can significantly reduce snake-venom-induced pathology in mice, at least in part by releasing carboxypeptidase A and possibly other proteases, which can degrade venom components. Mast cells also significantly reduced the morbidity and mortality induced by honeybee venom. These findings identify a new biological function for mast cells in enhancing resistance to the morbidity and mortality induced by animal venoms.

    View details for DOI 10.1126/science.1128877

    View details for Web of Science ID 000239308600064

    View details for PubMedID 16873664

  • Mast cells can promote the development of multiple features of chronic asthma in mice JOURNAL OF CLINICAL INVESTIGATION Yu, M., Tsai, M., Tam, S., Jones, C., Zehnder, J., Galli, S. J. 2006; 116 (6): 1633-1641

    Abstract

    Bronchial asthma, the most prevalent cause of significant respiratory morbidity in the developed world, typically is a chronic disorder associated with long-term changes in the airways. We developed a mouse model of chronic asthma that results in markedly increased numbers of airway mast cells, enhanced airway responses to methacholine or antigen, chronic inflammation including infiltration with eosinophils and lymphocytes, airway epithelial goblet cell hyperplasia, enhanced expression of the mucin genes Muc5ac and Muc5b, and increased levels of lung collagen. Using mast cell-deficient (Kit(W-sh/W-sh) and/or Kit(W/W-v)) mice engrafted with FcRgamma+/+ or FcRgamma-/- mast cells, we found that mast cells were required for the full development of each of these features of the model. However, some features also were expressed, although usually at less than wild-type levels, in mice whose mast cells lacked FcRgamma and therefore could not be activated by either antigen- and IgE-dependent aggregation of Fc epsilonRI or the binding of antigen-IgG1 immune complexes to Fc gammaRIII. These findings demonstrate that mast cells can contribute to the development of multiple features of chronic asthma in mice and identify both Fc Rgamma-dependent and Fc Rgamma-independent pathways of mast cell activation as important for the expression of key features of this asthma model.

    View details for DOI 10.1172/JCI25702

    View details for Web of Science ID 000237979700025

    View details for PubMedID 16710480

  • Mast cell-associated TNF promotes dendritic cell migration JOURNAL OF IMMUNOLOGY Suto, H., Nakae, S., Kakurai, M., Sedgwick, J. D., Tsai, M., Galli, S. J. 2006; 176 (7): 4102-4112

    Abstract

    Mast cells represent a potential source of TNF, a mediator which can enhance dendritic cell (DC) migration. Although the importance of mast cell-associated TNF in regulating DC migration in vivo is not clear, mast cells and mast cell-derived TNF can contribute to the expression of certain models of contact hypersensitivity (CHS). We found that CHS to FITC was significantly impaired in mast cell-deficient Kit(W-sh/W-sh) or TNF(-/)(-) mice. The reduced expression of CHS in Kit(W-sh/W-sh) mice was fully repaired by local transfer of wild-type bone marrow-derived cultured mast cells (BMCMCs), but was only partially repaired by transfer of TNF(-/)(-) BMCMCs. Thus, mast cells, and mast cell-derived TNF, were required for optimal expression of CHS to FITC. We found that the migration of FITC-bearing skin DCs into draining lymph nodes (LNs) 24 h after epicutaneous administration of FITC in naive mice was significantly reduced in mast cell-deficient or TNF(-/)(-) mice, but levels of DC migration in these mutant mice increased to greater than wild-type levels by 48 h after FITC sensitization. Mast cell-deficient or TNF(-/)(-) mice also exhibited significantly reduced migration of airway DCs to local LNs at 24 h after intranasal challenge with FITC-OVA. Migration of FITC-bearing DCs to LNs draining the skin or airways 24 h after sensitization was repaired in Kit(W-sh/W-sh) mice which had been engrafted with wild-type but not TNF(-/)(-) BMCMCs. Our findings indicate that mast cell-associated TNF can contribute significantly to the initial stages of FITC-induced migration of cutaneous or airway DCs.

    View details for Web of Science ID 000238769300035

    View details for PubMedID 16547246

  • RabGEF1 regulates stem cell factor/c-Kit-mediated signaling events and biological responses in mast cells PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Kalesnikoff, J., Rios, E. J., Chen, C. C., Nakae, S., Zabel, B. A., BUTCHER, E. C., Tsai, M., Tam, S. Y., Galli, S. J. 2006; 103 (8): 2659-2664

    Abstract

    We recently reported that RabGEF1 is a negative regulator of high-affinity Fc receptor for IgE (Fc epsilonRI)-dependent mast cell activation and that mice lacking RabGEF1 develop severe skin inflammation and increased numbers of dermal mast cells. To better understand how RabGEF1 can regulate signaling events and biological responses in mast cells, we examined the responses of bone marrow-derived cultured mast cells (BMCMCs) from wild-type (+/+) and Rabgef1 knockout (-/-) mice after stimulation with the c-Kit ligand, stem cell factor (SCF), an important regulator of mast cell development, survival, proliferation, and activation. We found that RabGEF1-deficient mast cells exhibited enhanced and prolonged activation of Ras and extracellular regulated kinase, and significantly elevated IL-6 secretion, after stimulation with SCF. SCF-induced activation of c-Jun N-terminal kinase was increased in Rabgef1-/- BMCMCs, but without corresponding significant increases in SCF-induced migration or adhesion. SCF-mediated activation of the survival-enhancing kinase, Akt, also was increased in Rabgef1-/- BMCMCs, and these cells had a survival advantage over their +/+ counterparts in vitro. Despite enhanced Ras activation in the absence of RabGEF1, SCF-induced proliferation was lower in Rabgef1-/- BMCMCs compared with their +/+ counterparts. Finally, we found that c-Kit internalization was delayed in the absence of RabGEF1, probably reflecting a positive role for RabGEF1 in the regulation of endocytic events, and that infection of Rabgef1-/- BMCMCs with a wild-type RabGEF1 lentiviral construct normalized c-Kit internalization to the levels seen in +/+ BMCMCs. Thus, RabGEF1 plays a critical role in the regulation of SCF/c-Kit-mediated signaling events and biological responses in mast cells.

    View details for Web of Science ID 000235554900034

    View details for PubMedID 16533754

  • Mast cells enhance T cell activation: Importance of mast cell costimulatory molecules and secreted TNF JOURNAL OF IMMUNOLOGY Nakae, S., Suto, H., Iikura, M., Kakurai, M., Sedgwick, J. D., Tsai, M., Galli, S. J. 2006; 176 (4): 2238-2248

    Abstract

    We recently reported that mast cells stimulated via FcepsilonRI aggregation can enhance T cell activation by a TNF-dependent mechanism. However, the molecular mechanisms responsible for such IgE-, Ag- (Ag-), and mast cell-dependent enhancement of T cell activation remain unknown. In this study we showed that mouse bone marrow-derived cultured mast cells express various costimulatory molecules, including members of the B7 family (ICOS ligand (ICOSL), PD-L1, and PD-L2) and the TNF/TNFR families (OX40 ligand (OX40L), CD153, Fas, 4-1BB, and glucocorticoid-induced TNFR). ICOSL, PD-L1, PD-L2, and OX40L also are expressed on APCs such as dendritic cells and can modulate T cell function. We found that IgE- and Ag-dependent mast cell enhancement of T cell activation required secreted TNF; that TNF can increase the surface expression of OX40, ICOS, PD-1, and other costimulatory molecules on CD3(+) T cells; and that a neutralizing Ab to OX40L, but not neutralizing Abs to ICOSL or PD-L1, significantly reduced IgE/Ag-dependent mast cell-mediated enhancement of T cell activation. These results indicate that the secretion of soluble TNF and direct cell-cell interactions between mast cell OX40L and T cell OX40 contribute to the ability of IgE- and Ag-stimulated mouse mast cells to enhance T cell activation.

    View details for Web of Science ID 000235180900025

    View details for PubMedID 16455980

  • A key regulatory role for histamine in experimental autoimmune encephalomyelitis: Disease exacerbation in histidine decarboxylase-deficient mice JOURNAL OF IMMUNOLOGY Musio, S., Gallo, B., Scabeni, S., Lapilla, M., Poliani, P. L., Matarese, G., Ohtsu, H., GALLI, S. J., Mantegazza, R., Steinman, L., Pedotti, R. 2006; 176 (1): 17-26

    Abstract

    Histamine can modulate the cytokine network and influence Th1 and Th2 balance and Ab-isotype switching. Thus, pharmacological blockade or genetic deletion of specific histamine receptors has been shown to reduce the severity of experimental autoimmune encephalomyelitis (EAE), a prototypic Th1-mediated disease with similarities to human multiple sclerosis. To study the comprehensive contribution of endogenous histamine to the expression of EAE, we attempted to induce EAE in histidine decarboxylase-deficient mice, which are genetically unable to make histamine. In this study, we show that EAE is significantly more severe in HDC-/-, histamine-deficient mice, with diffuse inflammatory infiltrates, including a prevalent granulocytic component, in the brain and cerebellum. Unlike splenocytes from wild-type mice, splenocytes from HDC-/- mice do not produce histamine in response to the myelin Ag, whereas production of IFN-gamma, TNF, and leptin are increased in HDC-/- splenocytes in comparison to those from wild-type mice. Endogenous histamine thus appears to regulate importantly the autoimmune response against myelin and the expression of EAE, in this model, and to limit immune damage to the CNS. Understanding which receptor(s) for histamine is/are involved in regulating autoimmunity against the CNS might help in the development of new strategies of treatment for EAE and multiple sclerosis.

    View details for Web of Science ID 000234262600005

    View details for PubMedID 16365391

  • Monomeric IgE enhances human mast cell chemokine production: ILA-4 augments and dexamethasone suppresses the response JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY Matsuda, K., Piliponsky, A. M., Iikura, M., Nakae, S., Wang, E. W., Dutta, S. M., Kawakami, T., Tsai, M., Galli, S. J. 2005; 116 (6): 1357-1363

    Abstract

    Mouse monoclonal IgE antibodies can promote the survival of mouse bone marrow-derived cultured mast cells and induce the cells to secrete mediators in the absence of known specific antigen.To determine whether human IgE, in the absence of known specific antigen, had effects on the mediator secretion or survival of human mast cells.We tested whether human IgE induced human cord blood-derived mast cells to secrete mediators or enhanced their survival on withdrawal of stem cell factor.Exposure to IgE, but not IgG, at concentrations as low as 2.5 microg/mL significantly enhanced the release of IL-8 and monocyte chemoattractant protein 1, but not histamine or cysteinyl leukotrienes. However, under the conditions tested, chemokine production in response to IgE alone was significantly less than that induced when aliquots of the same IgE-sensitized populations of human mast cells were stimulated with anti-IgE. The production of IL-8 and monocyte chemoattractant protein 1 in response to either IgE alone or IgE and anti-IgE was enhanced by preincubation of the cells in IL-4 and was inhibited by preincubation of the cells with dexamethasone. By contrast, we did not detect any ability of IgE to enhance mast cell survival on withdrawal of stem cell factor.Exposure to human IgE in vitro in the absence of known specific antigen can enhance chemokine production by human mast cells, and this secretory response can be enhanced by preincubation of the mast cells with IL-4 and can be suppressed by dexamethasone.

    View details for DOI 10.1016/j.jaci.2005.08.042

    View details for Web of Science ID 000235687000030

    View details for PubMedID 16337471

  • Activity of the tyrosine kinase inhibitor PKC412 in a patient with mast cell leukemia with the D816V KIT mutation BLOOD Gotlib, J., Berube, C., Growney, J. D., Chen, C. C., George, T. I., Williams, C., Kajiguchi, T., Ruan, J., Lilleberg, S. L., Durocher, J. A., Lichy, J. H., Wang, Y. F., Cohen, P. S., Arber, D. A., Heinrich, M. C., Neckers, L., GALLI, S. J., Gilliland, D. G., Coutre, S. E. 2005; 106 (8): 2865-2870

    Abstract

    The majority of patients with systemic mast cell disease express the imatinib-resistant Asp816Val (D816V) mutation in the KIT receptor tyrosine kinase. Limited treatment options exist for aggressive systemic mastocytosis (ASM) and mast cell leukemia (MCL). We evaluated whether PKC412, a small-molecule inhibitor of KIT with a different chemical structure from imatinib, may have therapeutic use in advanced SM with the D816V KIT mutation. We treated a patient with MCL (with an associated myelodysplastic syndrome (MDS)/myeloproliferative disorder [MPD]) based on in vitro studies demonstrating that PKC412 could inhibit D816V KIT-transformed Ba/F3 cell growth with a 50% inhibitory concentration (IC50) of 30 nM to 40 nM. The patient exhibited a partial response with significant resolution of liver function abnormalities. In addition, PKC412 treatment resulted in a significant decline in the percentage of peripheral blood mast cells and serum histamine level and was associated with a decrease in KIT phosphorylation and D816V KIT mutation frequency. The patient died after 3 months of therapy due to progression of her MDS/MPD to acute myeloid leukemia (AML). This case indicates that KIT tyrosine kinase inhibition is a feasible approach in SM, but single-agent clinical efficacy may be limited by clonal evolution in the advanced leukemic phase of this disease.

    View details for DOI 10.1182/blood-2005-04-1568

    View details for Web of Science ID 000232466000047

    View details for PubMedID 15972446

  • Mast cell-deficient W-sash c-kit mutant Kit(W-sh/W-sh) mice as a model for investigating mast cell biology in vivo AMERICAN JOURNAL OF PATHOLOGY Grimbaldeston, M. A., Chen, C. C., Piliponsky, A. M., Tsai, M., Tam, S. Y., Galli, S. J. 2005; 167 (3): 835-848

    Abstract

    Mice carrying certain mutations in the white spotting (W) locus (ie, c-kit) exhibit reduced c-kit tyrosine kinase-dependent signaling that results in mast cell deficiency and other phenotypic abnormalities. The c-kit mutations in Kit(W/W-v) mice impair melanogenesis and result in anemia, sterility, and markedly reduced levels of tissue mast cells. In contrast, Kit(W-sh/W-sh) mice, bearing the W-sash (W(sh)) inversion mutation, have mast cell deficiency but lack anemia and sterility. We report that adult Kit(W-sh/W-sh) mice had a profound deficiency in mast cells in all tissues examined but normal levels of major classes of other differentiated hematopoietic and lymphoid cells. Unlike Kit(W/W-v) mice, Kit(W-sh/W-sh) mice had normal numbers of TCR gammadelta intraepithelial lymphocytes in the intestines and did not exhibit a high incidence of idiopathic dermatitis, ulcers, or squamous papillomas of the stomach, but like Kit(W/W-v) mice, they lacked interstitial cells of Cajal in the gut and exhibited bile reflux into the stomach. Systemic or local reconstitution of mast cell populations was achieved in nonirradiated adult Kit(W-sh/W-sh) mice by intravenous, intraperitoneal, or intradermal injection of wild-type bone marrow-derived cultured mast cells but not by transplantation of wild-type bone marrow cells. Thus, Kit(W-sh/W-sh) mice represent a useful model for mast cell research, especially for analyzing mast cell function in vivo.

    View details for Web of Science ID 000231514500018

    View details for PubMedID 16127161

  • Identification of mast cell progenitors in adult mice PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Chen, C. C., Grimbaldeston, M. A., Tsai, M., Weissman, I. L., Galli, S. J. 2005; 102 (32): 11408-11413

    Abstract

    It is well known that mast cells are derived from hematopoietic stem cells. However, in adult hematopoiesis, a committed mast cell progenitor has not yet been identified in any species, nor is it clear at what point during adult hematopoiesis commitment to the mast cell lineage occurs. We identified a cell population in adult mouse bone marrow, characterized as Lin(-)c-Kit(+)Sca-1(-)-Ly6c(-)FcepsilonRIalpha(-)CD27(-)beta7(+)T1/ST2+, that gives rise only to mast cells in culture and that can reconstitute the mast cell compartment when transferred into c-kit mutant mast cell-deficient mice. In addition, our experiments strongly suggest that these adult mast cell progenitors are derived directly from multipotential progenitors instead of, as previously proposed, common myeloid progenitors or granulocyte/macrophage progenitors.

    View details for DOI 10.1073/pnas.0504197102

    View details for Web of Science ID 000231253400051

    View details for PubMedID 16006518

  • Mast cells enhance T cell activation: Importance of mast cell-derived TNF PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Nakae, S., Suto, H., Kakurai, M., Sedgwick, J. D., Tsai, M., Galli, S. J. 2005; 102 (18): 6467-6472

    Abstract

    Mast cells are not only important effector cells in immediate hypersensitivity reactions and immune responses to pathogens but also can contribute to T cell-mediated disorders. However, the mechanisms by which mast cells might influence T cells in such settings are not fully understood. We find that mast cells can enhance proliferation and cytokine production in multiple T cell subsets. Mast cell-dependent enhancement of T cell activation can be promoted by FcepsilonRI-dependent mast cell activation, TNF production by both mast cells and T cells, and mast cell-T cell contact. However, at high concentrations of cells, mast cells can promote T cell activation independent of IgE or TNF. Finally, mast cells also can promote T cell activation by means of soluble factors. These findings identify multiple mechanisms by which mast cells can influence T cell proliferation and cytokine production.

    View details for DOI 10.1073/pnas.0501912102

    View details for Web of Science ID 000228918400041

    View details for PubMedID 15840716

  • Pathogenesis and management of anaphylaxis: Current status and future challenges JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY Galli, S. J. 2005; 115 (3): 571-574

    View details for DOI 10.1016/j.jaci.2004.12.1133

    View details for Web of Science ID 000227687000021

    View details for PubMedID 15753906

  • Decreased susceptibility of mast cell-deficient Kit(W)/Kit(W-v) mice to the development of 1,2-dimethylhydrazine-induced intestinal tumors LABORATORY INVESTIGATION Wedemeyer, J., Galli, S. J. 2005; 85 (3): 388-396

    Abstract

    Administration of 1,2-dimethylhydrazine (DMH) induces intestinal epithelial tumors in mice. Increased numbers of mast cells have been reported to occur both within and near a variety of different neoplasms, including DMH-induced intestinal tumors. We investigated the role of the tyrosine kinase receptor, c-kit, and mast cells, in this model by administering DMH to c-kit mutant mast cell-deficient mice and the congenic normal mice. We attempted to induce colonic tumors by administering DMH (20 mg/kg body weight, s.c., weekly for 20 weeks) to WBB6F1-Kit+/+ (+/+) wild-type mice, the congenic mast cell-deficient WBB6F1-Kit(W)/Kit(W-v) (W/W(v)) mice and W/W(v) mice that had been repaired of their mast cell deficiency by adoptive transfer of bone marrow cells derived from the congenic +/+ mice. The susceptibility to the development of DMH-induced colonic tumors, and the numbers of mast cells associated with these tumors, was evaluated. Normal (+/+) mice exhibited significantly higher numbers of mast cells in DMH-induced intestinal tumors than in macroscopically normal colonic mucosa. Treatment with DMH induced development of colonic tumors in 97% of +/+ mice, but in only 32% of the W/W(v) mice. W/W(v) mice that had been repaired of their mast cell deficiency by transfer of +/+ bone marrow cells expressed susceptibility to the development of colonic tumors that was similar to that of wild-type mice. These results show that genetic impairment of c-kit function reduces the susceptibility of mice to DMH-induced colonic tumors, and that defects in bone marrow-derived cells in the W/W(v) mice contribute significantly to this result. Our findings also are consistent with the possibility that mast cells promote the development of DMH-induced colonic epithelial tumors in mice.

    View details for DOI 10.1038/labinvest.3700232

    View details for Web of Science ID 000227168700009

    View details for PubMedID 15696191

  • Symposium on the definition and management of anaphylaxis: Summary report JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY Sampson, H. A., Munoz-Furlong, A., Bock, A., Schmitt, M., Bass, R., Chowdhury, B. A., Decker, W. W., Furlong, T. J., GALLI, S. J., Golden, D. B., Gruchalla, R. S., Harlor, A. D., Hepner, D. L., Howarth, M., Kaplan, A. P., Levy, J. H., Lewis, L. M., Lieberman, P. L., Metcalfe, D. D., Murphy, R., Pollart, S. M., Pumphrey, R. S., Rosenwasser, L. J., Simons, F. E., Wood, J. P., Camargo, C. A. 2005; 115 (3): 584-591

    View details for DOI 10.1016/j.jaci.2005.01.008

    View details for Web of Science ID 000227687000023

    View details for PubMedID 15753908

  • Mast cells in the development of adaptive immune responses NATURE IMMUNOLOGY Galli, S. J., Nakae, S., Tsai, M. 2005; 6 (2): 135-142

    Abstract

    Mast cells are so widely recognized as critical effector cells in allergic disorders and other immunoglobulin E-associated acquired immune responses that it can be difficult to think of them in any other context. However, mast cells also can be important as initiators and effectors of innate immunity. In addition, mast cells that are activated during innate immune responses to pathogens, or in other contexts, can secrete products and have cellular functions with the potential to facilitate the development, amplify the magnitude or regulate the kinetics of adaptive immune responses. Thus, mast cells may influence the development, intensity and duration of adaptive immune responses that contribute to host defense, allergy and autoimmunity, rather than simply functioning as effector cells in these settings.

    View details for DOI 10.1038/ni1158

    View details for Web of Science ID 000226468100016

    View details for PubMedID 15662442

  • Mast cells as "tunable" effector and immunoregulatory cells: Recent advances ANNUAL REVIEW OF IMMUNOLOGY Galli, S. J., Kalesnikoff, J., Grimbaldeston, M. A., Piliponsky, A. M., Williams, C. M., Tsai, M. 2005; 23: 749-786

    Abstract

    This review focuses on recent progress in our understanding of how mast cells can contribute to the initiation, development, expression, and regulation of acquired immune responses, both those associated with IgE and those that are apparently expressed independently of this class of Ig. We emphasize findings derived from in vivo studies in mice, particularly those employing genetic approaches to influence mast cell numbers and/or to alter or delete components of pathways that can regulate mast cell development, signaling, or function. We advance the hypothesis that mast cells not only can function as proinflammatory effector cells and drivers of tissue remodeling in established acquired immune responses, but also may contribute to the initiation and regulation of such responses. That is, we propose that mast cells can also function as immunoregulatory cells. Finally, we show that the notion that mast cells have primarily two functional configurations, off (or resting) or on (or activated for extensive mediator release), markedly oversimplifies reality. Instead, we propose that mast cells are "tunable," by both genetic and environmental factors, such that, depending on the circumstances, the cell can be positioned phenotypically to express a wide spectrum of variation in the types, kinetics, and/or magnitude of its secretory functions.

    View details for DOI 10.1146/annurev.immunol.21.120601.141025

    View details for Web of Science ID 000228947000022

    View details for PubMedID 15771585

  • RabGEF1, a negative regulator of Ras signalling, mast cell activation and skin inflammation. Novartis Foundation symposium Tam, S., Kalesnikoff, J., Nakae, S., Tsai, M., Galli, S. J. 2005; 271: 115-124

    Abstract

    Mast cell activation induced by the aggregation of FcepsilonRI with IgE and antigen is mediated through the activation of multiple protein kinase cascades. This process induces mast cells to undergo degranulation, to synthesize and release lipid mediators, and to secrete multiple cytokines, chemokines and growth factors. We found that RabGEF1 (Rabex-5) binds to Ras and negatively regulates Ras activation and downstream effector pathways during FcepsilonRI-dependent mouse mast cell activation. Mast cells derived from RabGEF1-deficient mice exhibit significantly enhanced levels of degranulation, release of lipid mediators and secretion of cytokines in response to FcepsilonRI aggregation. RabGEF1 knockout mice have increased perinatal mortality and the mice that do survive develop severe skin inflammation and increased numbers of mast cells in the dermis, some of which exhibit morphological evidence of degranulation. These mice also show elevated concentrations of serum histamine and IgE. Thus, RabGEF1 is a negative regulator of Ras signalling and FcepsilonRI-dependent mast cell activation in vitro, and a lack of RabGEF1 results in the development of elevated numbers of mast cells in the skin and severe skin inflammation in vivo.

    View details for PubMedID 16605131

  • Mast cells promote homeostasis by limiting endothelin-1-induced toxicity NATURE Maurer, M., Wedemeyer, J., Metz, M., Piliponsky, A. M., Weller, K., Chatterjea, D., Clouthier, D. E., Yanagisawa, M. M., Tsai, M., Galli, S. J. 2004; 432 (7016): 512-516

    Abstract

    Endothelin-1 (ET-1) is a 21-amino-acid peptide, derived from vascular endothelial cells, with potent vasoconstrictor activity. ET-1 has been implicated in diverse physiological or pathological processes, including the vascular changes associated with sepsis. However, the factors that regulate ET-1-associated toxicity during bacterial infections, or in other settings, are not fully understood. Both the pathology associated with certain allergic and autoimmune disorders, and optimal host defence against bacterial and parasitic infections are mediated by mast cells. In vitro, mast cells can produce ET-1 (ref. 11), undergo ET-1-dependent and endothelin-A receptor (ET(A))-dependent activation, and release proteases that degrade ET-1 (ref. 14). Although the potential relationships between mast cells and the ET-1 system thus may be complex, the importance of interactions between ET-1 and mast cells in vivo is obscure. Here we show that ET(A)-dependent mast-cell activation can diminish both ET-1 levels and ET-1-induced pathology in vivo, and also can contribute to optimal survival during acute bacterial peritonitis. These findings identify a new biological function for mast cells: promotion of homeostasis by limiting the toxicity associated with an endogenous mediator.

    View details for DOI 10.1038/nature03085

    View details for Web of Science ID 000225322100049

    View details for PubMedID 15543132

  • RabGEF1 is a negative regulator of mast cell activation and skin inflammation NATURE IMMUNOLOGY Tam, S. Y., Tsai, M., Snouwaert, J. N., Kalesnikoff, J., Scherrer, D., Nakae, S., Chatterjea, D., Bouley, D. M., Galli, S. J. 2004; 5 (8): 844-852

    Abstract

    Mast cell activation induced by aggregation of Fc epsilon RI receptors with immunoglobulin E and antigen is mediated through the activation of multiple protein kinase cascades. Here we report that the regulatory protein RabGEF1 bound to Ras and negatively regulated Ras activation and its 'downstream' effector pathways in Fc epsilon RI-dependent mast cell activation. RabGEF1-deficient mast cells showed enhanced degranulation and release of lipid mediators and cytokines in response to Fc epsilon RI aggregation. RabGEF1-deficient mice developed severe skin inflammation and had increased numbers of mast cells. Thus, RabGEF1 is a negative regulator of Fc epsilon RI-dependent mast cell activation, and a lack of RabGEF1 results in the development of skin inflammation in vivo.

    View details for DOI 10.1038/ni1093

    View details for Web of Science ID 000222955600016

    View details for PubMedID 15235600

  • Immune sensitization in the skin is enhanced by antigen-independent effects of IgE IMMUNITY Bryce, P. J., Miller, M. L., Miyajima, I., Tsai, M., GALLI, S. J., Oettgen, H. C. 2004; 20 (4): 381-392

    Abstract

    Contact sensitivity responses require both effective immune sensitization following cutaneous exposure to chemical haptens and antigen-specific elicitation of inflammation upon subsequent hapten challenge. We report that antigen-independent effects of IgE antibodies can promote immune sensitization to haptens in the skin. Contact sensitivity was markedly impaired in IgE(-/-) mice but was restored by either transfer of sensitized cells from wild-type mice or administration of hapten-irrelevant IgE before sensitization. Moreover, IgE(-/-) mice exhibited impairment in the reduction of dendritic cell numbers in the epidermis after hapten exposure. Monomeric IgE has been reported to influence mast cell function. We observed diminished contact sensitivity in mice lacking FcepsilonRI or mast cells, and mRNA for several mast cell-associated genes was reduced in IgE(-/-) versus wild-type skin after hapten exposure. We speculate that levels of IgE normally present in mice favor immune sensitization via antigen-independent but FcepsilonRI-dependent effects on mast cells.

    View details for Web of Science ID 000221442800005

    View details for PubMedID 15084268

  • Chair's introduction. Anaphylaxis. Novartis Foundation symposium Galli, S. J. 2004; 257: 1-5

    View details for PubMedID 15025388

  • Evidence that IgE molecules mediate a spectrum of effects on mast cell survival and activation via aggregation of the Fc epsilon RI PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Kitaura, J., Song, J. M., Tsai, M., Asai, K., Maeda-Yamamoto, M., Mocsai, A., Kawakami, Y., Liu, F. T., Lowell, C. A., Barisas, B. G., GALLI, S. J., Kawakami, T. 2003; 100 (22): 12911-12916

    Abstract

    We demonstrate that binding of different IgE molecules (IgEs) to their receptor, FcepsilonRI, induces a spectrum of activation events in the absence of a specific antigen and provide evidence that such activation reflects aggregation of FcepsilonRI. Highly cytokinergic IgEs can efficiently induce production of cytokines and render mast cells resistant to apoptosis in an autocrine fashion, whereas poorly cytokinergic IgEs induce these effects inefficiently. Highly cytokinergic IgEs seem to induce more extensive FcepsilonRI aggregation than do poorly cytokinergic IgEs, which leads to stronger mast cell activation and survival effects. These effects of both types of IgEs require Syk tyrosine kinase and can be inhibited by FcepsilonRI disaggregation with monovalent hapten. In hybridoma-transplanted mice, mucosal mast cell numbers correlate with serum IgE levels. Therefore, survival effects of IgE could contribute to the pathogenesis of allergic disease.

    View details for DOI 10.1073/pnas.1735525100

    View details for Web of Science ID 000186301100073

    View details for PubMedID 14569021

  • Involvement of both 'allergic' and "autoimmune' mechanisms in EAE, MS and other autoimmune diseases TRENDS IN IMMUNOLOGY Pedotti, R., De Voss, J. J., Steinman, L., GALLI, S. J. 2003; 24 (9): 479-484
  • Identification of A(3) receptor- and mast cell-dependent and -independent components of adenosine-mediated airway responsiveness in mice JOURNAL OF IMMUNOLOGY Tilley, S. L., Tsai, M., Williams, C. M., Wang, Z. S., Erikson, C. J., GALLI, S. J., Koller, B. H. 2003; 171 (1): 331-337

    Abstract

    Adenosine-induced bronchoconstriction is a well-recognized feature of atopic asthma. Adenosine acts through four different G protein-coupled receptors to produce a myriad of physiological effects. To examine the contribution of the A(3) adenosine receptor to adenosine-induced bronchoconstriction and to assess the contribution of mast cells to this process, we quantified airway responsiveness to aerosolized adenosine in wild-type, A(3) receptor-deficient, and mast cell-deficient mice. Compared with the robust airway responses elicited by adenosine in wild-type mice, both A(3)-deficient and mast cell-deficient mice exhibited a significantly attenuated response compared with their respective wild-type controls. Histological examination of the airways 4 h after adenosine exposure revealed extensive degranulation of airway mast cells as well as infiltration of neutrophils in wild-type mice, whereas these findings were much diminished in A(3)-deficient mice and were not different from those in PBS-treated controls. These data indicate that the airway responses to aerosolized adenosine in mice occur largely through A(3) receptor activation and that mast cells contribute significantly to these responses, but that activation of additional adenosine receptors on a cell type(s) other than mast cells also contributes to adenosine-induced airway responsiveness in mice. Finally, our findings indicate that adenosine exposure can result in A(3)-dependent airway inflammation, as reflected in neutrophil recruitment, as well as alterations in airway function.

    View details for Web of Science ID 000183674400042

    View details for PubMedID 12817015

  • Severe anaphylactic reactions to glutamic acid decarboxylase (GAD) self peptides in NOD mice that spontaneously develop autoimmune type 1 diabetes mellitus. BMC immunology Pedotti, R., Sanna, M., Tsai, M., DeVoss, J., Steinman, L., McDevitt, H., Galli, S. J. 2003; 4: 2-?

    Abstract

    Insulin dependent (i.e., "type 1") diabetes mellitus (T1DM) is considered to be a T cell mediated disease in which TH1 and Tc autoreactive cells attack the pancreatic islets. Among the beta-cell antigens implicated in T1DM, glutamic acid decarboxylase (GAD) 65 appears to play a key role in the development of T1DM in humans as well as in non-obese diabetic (NOD) mice, the experimental model for this disease. It has been shown that shifting the immune response to this antigen from TH1 towards TH2, via the administration of GAD65 peptides to young NOD mice, can suppress the progression to overt T1DM. Accordingly, various protocols of "peptide immunotherapy" of T1DM are under investigation. However, in mice with experimental autoimmune encephalomyelitis (EAE), another autoimmune TH1 mediated disease that mimics human multiple sclerosis, anaphylactic shock can occur when the mice are challenged with certain myelin self peptides that initially were administered with adjuvant to induce the disease.Here we show that NOD mice, that spontaneously develop T1DM, can develop fatal anaphylactic reactions upon challenge with preparations of immunodominant GAD65 self peptides after immunization with these peptides to modify the development of T1DM.These findings document severe anaphylaxis to self peptide preparations used in an attempt to devise immunotherapy for a spontaneous autoimmune disease. Taken together with the findings in EAE, these results suggest that peptide therapies designed to induce a TH1 to TH2 shift carry a risk for the development of anaphylactic reactivity to the therapeutic peptides.

    View details for PubMedID 12597780

  • Multiple elements of the allergic arm of the immune response modulate autoimmune demyelination PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Pedotti, R., DeVoss, J. J., Youssef, S., Mitchell, D., Wedemeyer, J., Madanat, R., Garren, H., Fontoura, P., Tsai, M., GALLI, S. J., Sobel, R. A., Steinman, L. 2003; 100 (4): 1867-1872

    Abstract

    Analysis of mRNA from multiple sclerosis lesions revealed increased amounts of transcripts for several genes encoding molecules traditionally associated with allergic responses, including prostaglandin D synthase, histamine receptor type 1 (H1R), platelet activating factor receptor, Ig Fc epsilon receptor 1 (Fc epsilon RI), and tryptase. We now demonstrate that, in the animal model of multiple sclerosis, experimental autoimmune encephalomyelitis (EAE), mediated by T helper 1 (Th1) T cells, histamine receptor 1 and 2 (H1R and H2R) are present on inflammatory cells in brain lesions. Th1 cells reactive to myelin proteolipid protein expressed more H1R and less H2R than Th2 cells. Pyrilamine, an H1R antagonist, blocked EAE, and the platelet activating factor receptor antagonist CV6209 reduced the severity of EAE. EAE severity was also decreased in mice with disruption of the genes encoding Ig Fc gamma RIII or both Fc gamma RIII and Fc epsilon RI. Prostaglandin D synthase and tryptase transcripts were elevated in EAE brain. Taken together, these data reveal extensive involvement of elements of the immune response associated with allergy in autoimmune demyelination. The pathogenesis of demyelination must now be viewed as encompassing elements of both Th1 responses and "allergic" responses.

    View details for DOI 10.1073/pnas.252777399

    View details for Web of Science ID 000181073000077

    View details for PubMedID 12576552

  • Regulation of mast-cell and basophil function and survival by IgE NATURE REVIEWS IMMUNOLOGY Kawakami, T., Galli, S. J. 2002; 2 (10): 773-786

    Abstract

    Mast cells and basophils are important effector cells in T helper 2 (T(H)2)-cell-dependent, immunoglobulin-E-associated allergic disorders and immune responses to parasites. The crosslinking of IgE that is bound to the high-affinity receptor Fc epsilon RI with multivalent antigen results in the aggregation of Fc epsilon RI and the secretion of products that can have effector, immunoregulatory or autocrine effects. This response can be enhanced markedly in cells that have been exposed to high levels of IgE, which results in the increased surface expression of Fc epsilon RI. Moreover, recent work indicates that monomeric IgE (in the absence of crosslinking) can render mast cells resistant to apoptosis induced by growth-factor deprivation in vitro and, under certain circumstances, can induce the release of cytokines. So, the binding of IgE to Fc epsilon RI might influence mast-cell and basophil survival directly or indirectly, and can also regulate cellular function.

    View details for DOI 10.1038/nri914

    View details for Web of Science ID 000180439500021

    View details for PubMedID 12360215

  • Transcriptional response of human mast cells stimulated via the Fc(epsilon)RI and identification of mast cells as a source of IL-11. BMC immunology Sayama, K., Diehn, M., Matsuda, K., Lunderius, C., Tsai, M., Tam, S., Botstein, D., Brown, P. O., Galli, S. J. 2002; 3: 5-?

    Abstract

    In asthma and other allergic disorders, the activation of mast cells by IgE and antigen induces the cells to release histamine and other mediators of inflammation, as well as to produce certain cytokines and chemokines. To search for new mast cell products, we used complementary DNA microarrays to analyze gene expression in human umbilical cord blood-derived mast cells stimulated via the high-affinity IgE receptor (Fc(epsilon)RI).One to two hours after Fc(epsilon)RI-dependent stimulation, more than 2,400 genes (about half of which are of unknown function) exhibited 2-200 fold changes in expression. The transcriptional program included changes in the expression of IL-11 and at least 30 other cytokines and chemokines. Human mast cells secreted 130-529 pg of IL-11/106 cells by 6 h after stimulation with anti-IgE.Our initial analysis of the transcriptional program induced in in vitro-derived human mast cells stimulated via the Fc(epsilon)RI has identified many products that heretofore have not been associated with this cell type, but which may significantly influence mast cell function in IgE-associated host responses. We also have demonstrated that mast cells stimulated via the Fc(epsilon)RI can secrete IL-11. Based on the previously reported biological effects of IL-11, our results suggest that production of IL-11 may represent one link between IgE-dependent mast cell activation in subjects with allergic asthma and the development of a spectrum of structural changes in the airways of these individuals; such changes, collectively termed "airway remodeling," can constitute an important long term consequence of asthma.

    View details for PubMedID 12079505

  • Gene-microarray analysis of multiple sclerosis lesions yields new targets validated in autoimmune encephalomyelitis NATURE MEDICINE Lock, C., Hermans, G., Pedotti, R., Brendolan, A., Schadt, E., Garren, H., Langer-Gould, A., Strober, S., Cannella, B., Allard, J., Klonowski, P., Austin, A., Lad, N., Kaminski, N., GALLI, S. J., Oksenberg, J. R., Raine, C. S., Heller, R., Steinman, L. 2002; 8 (5): 500-508

    Abstract

    Microarray analysis of multiple sclerosis (MS) lesions obtained at autopsy revealed increased transcripts of genes encoding inflammatory cytokines, particularly interleukin-6 and -17, interferon-gamma and associated downstream pathways. Comparison of two poles of MS pathology--acute lesions with inflammation versus 'silent' lesions without inflammation--revealed differentially transcribed genes. Some products of these genes were chosen as targets for therapy of experimental autoimmune encephalomyelitis (EAE) in mice. Granulocyte colony-stimulating factor is upregulated in acute, but not in chronic, MS lesions, and the effect on ameliorating EAE is more pronounced in the acute phase, in contrast to knocking out the immunoglobulin Fc receptor common gamma chain where the effect is greatest on chronic disease. These results in EAE corroborate the microarray studies on MS lesions. Large-scale analysis of transcripts in MS lesions elucidates new aspects of pathology and opens possibilities for therapy.

    View details for DOI 10.1038/nm0502-500

    View details for Web of Science ID 000175336800036

    View details for PubMedID 11984595

  • Regulation of mast cell survival by IgE IMMUNITY Asai, K., Kitaura, J., Kawakami, Y., Yamagata, N., Tsai, M., Carbone, D. P., Liu, F. T., GALLI, S. J., Kawakami, T. 2001; 14 (6): 791-800

    Abstract

    Mast cells play critical roles in hypersensitivity and in defense against certain parasites. We provide evidence that mouse mast cell survival and growth are promoted by monomeric IgE binding to its high-affinity receptor, Fc epsilon RI. Monomeric IgE does not promote DNA synthesis but suppresses the apoptosis induced by growth factor deprivation. This antiapoptotic effect occurs in parallel with IgE-induced increases in Fc epsilon RI surface expression but requires the continuous presence of IgE. This process does not involve the FasL/Fas death pathway or several Bcl-2 family proteins and induces a distinctly different signal than Fc epsilon RI cross-linking. The ability of IgE to enhance mast cell survival and Fc epsilon RI expression may contribute to amplified allergic reactions.

    View details for Web of Science ID 000169495100014

    View details for PubMedID 11420048

  • An unexpected version of horror autotoxicus: anaphylactic shock to a self peptide NATURE IMMUNOLOGY Pedotti, R., Mitchell, D., Wedemeyer, J., Karpuj, M., Chabas, D., Hattab, E. M., Tsai, M., GALLI, S. J., Steinman, L. 2001; 2 (3): 216-222

    Abstract

    EAE can refer either to experimental autoimmune encephalomyelitis or experimental allergic encephalomyelitis. Although EAE is classically a prototypic T helper 1 (TH1) cell-mediated autoimmune disease, it can also be induced by TH2 cells. Characteristically, the most severe manifestation of allergy, anaphylaxis, is associated with exposure to a foreign antigen that is often derived from medication, insect venom or food. We report here that, after self-tolerance to myelin is destroyed, anaphylaxis may be triggered by a self-antigen, in this case a myelin peptide. "Horror autotoxicus", which was initially described by Ehrlich, may not only include autoimmunity to self, it may also encompass immediate hypersensitivity to self, which leads to shock and rapid death.

    View details for Web of Science ID 000167413800013

    View details for PubMedID 11224520

  • Mast cells can amplify airway reactivity and features of chronic inflammation in an asthma model in mice JOURNAL OF EXPERIMENTAL MEDICINE Williams, C. M., Galli, S. J. 2000; 192 (3): 455-462

    Abstract

    The importance of mast cells in the development of the allergen-induced airway hyperreactivity and inflammation associated with asthma remains controversial. We found that genetically mast cell-deficient WBB6F(1)-W/W(v) mice that were sensitized to ovalbumin (OVA) without adjuvant, then challenged repetitively with antigen intranasally, exhibited much weaker responses in terms of bronchial hyperreactivity to aerosolized methacholine, lung tissue eosinophil infiltration, and numbers of proliferating cells within the airway epithelium than did identically treated WBB6F(1)-+/+ normal mice. However, W/W(v) mice that had undergone selective reconstitution of tissue mast cells with in vitro-derived mast cells of congenic +/+ mouse origin exhibited airway responses that were very similar to those of the +/+ mice. By contrast, W/W(v) mice that were sensitized with OVA emulsified in alum and challenged with aerosolized OVA exhibited levels of airway hyperreactivity and lung tissue eosinophil infiltration that were similar to those of the corresponding +/+ mice. Nevertheless, these W/W(v) mice exhibited significantly fewer proliferating cells within the airway epithelium than did identically treated +/+ mice. These results show that, depending on the "asthma model" investigated, mast cells can either have a critical role in, or not be essential for, multiple features of allergic airway responses in mice.

    View details for Web of Science ID 000088708800016

    View details for PubMedID 10934234

  • In vivo immunological function of mast cells derived from embryonic stem cells: An approach for the rapid analysis of even embryonic lethal mutations in adult mice in vivo PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Tsai, M., Wedemeyer, J., Ganiatsas, S., Tam, S. Y., Zon, L. I., GALLI, S. J. 2000; 97 (16): 9186-9190

    Abstract

    An important goal of tissue engineering is to achieve reconstitution of specific functionally active cell types by transplantation of differentiated cell populations derived from normal or genetically altered embryonic stem cells in vitro. We find that mast cells derived in vitro from wild-type or genetically manipulated embryonic stem cells can survive and orchestrate immunologically specific IgE-dependent reactions after transplantation into mast cell-deficient Kit(W)/Kit(W-v) mice. These findings define a unique approach for analyzing the effects of mutations of any genes that are expressed in mast cells, including embryonic lethal mutations, in vitro or in vivo.

    View details for Web of Science ID 000088608000077

    View details for PubMedID 10908668

  • A role for Bax in the regulation of apoptosis in mouse mast cells JOURNAL OF INVESTIGATIVE DERMATOLOGY Maurer, M., Tsai, M., Metz, M., Fish, S., Korsmeyer, S. J., GALLI, S. J. 2000; 114 (6): 1205-1206

    View details for Web of Science ID 000087365800022

    View details for PubMedID 10844568

  • The diverse potential effector and immunoregulatory roles of mast cells in allergic disease JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY Williams, C. M., GALLI, S. J. 2000; 105 (5): 847-859

    Abstract

    Mast cells are of hematopoietic origin but typically complete their maturation in peripheral connective tissues, especially those near epithelial surfaces. Mast cells express receptors that bind IgE antibodies with high affinity (FcepsilonRI), and aggregation of these FcepsilonRI by the reaction of cell-bound IgE with specific antigens induces mast cells to secrete a broad spectrum of biologically active preformed or lipid mediators, as well as many cytokines. Mast cells are widely thought to be essential for the expression of acute allergic reactions, but the importance of mast cells in late-phase reactions and chronic allergic inflammation has remained controversial. Although it is clear that many cell types may be involved in the expression of late-phase reactions and chronic allergic inflammation, studies in genetically mast cell-deficient and congenic normal mice indicate that mast cells may be critical for the full expression of certain features of late-phase reactions and may also contribute importantly to clinically relevant aspects of chronic allergic inflammation. Moreover, the pattern of cytokines that can be produced by mast cell populations, and the enhancement of such cytokine production in mast cells that have undergone IgE-dependent up-regulation of their surface expression of FcepsilonRI, suggests that mast cells may contribute to allergic diseases (and host defense) by acting as immunoregulatory cells, as well as by providing effector cell function.

    View details for Web of Science ID 000087185000001

    View details for PubMedID 10808163

  • Mast cells and basophils CURRENT OPINION IN HEMATOLOGY Galli, S. J. 2000; 7 (1): 32-39

    Abstract

    Mast cells and basophils are effector cells in IgE-associated immune responses, such as those that contribute to asthma and other allergic diseases and to host resistance to parasites. Recent work shows that mast cells can also participate in innate immunity to bacterial infection and that the expression of such mast cell-dependent natural immunity can be significantly enhanced by long-term treatment of mice with the kit ligand, stem cell factor. However, mast cells may also influence many other biologic responses, including tissue remodeling and angiogenesis. This review discusses certain recent findings about the differentiation, phenotype, and function of basophils and mast cells, as well as briefly considering evolving concepts about the roles of these cells in health and disease.

    View details for Web of Science ID 000165671100007

    View details for PubMedID 10608502

  • Mast-cell heparin demystified. Nature Zehnder, J. L., GALLI, S. J. 1999; 400 (6746): 714-715

    View details for PubMedID 10466718

  • IgE enhances Fc epsilon receptor I expression and IgE-dependent release of histamine and lipid mediators from human umbilical cord blood-derived mast cells: Synergistic effect of IL-4 and IgE on human mast cell Fc epsilon receptor I expression and mediator release JOURNAL OF IMMUNOLOGY Yamaguchi, M., Sayama, K., Yano, K., Lantz, C. S., Noben-Trauth, N., Ra, C., Costa, J. J., GALLI, S. J. 1999; 162 (9): 5455-5465

    Abstract

    We investigated the effects of IgE versus IL-4 on Fc epsilon RI surface expression in differentiated human mast cells derived in vitro from umbilical cord blood mononuclear cells. We found that IgE (at 5 micrograms/ml) much more strikingly enhanced surface expression of Fc epsilon RI than did IL-4 (at 0.1-100 ng/ml); similar results were also obtained with differentiated mouse mast cells. However, IL-4 acted synergistically with IgE to enhance Fc epsilon RI expression in these umbilical cord blood-derived human mast cells, as well as in mouse peritoneal mast cells derived from IL-4-/- or IL-4+/+ mice. We also found that: 1) IgE-dependent enhancement of Fc epsilon RI expression was associated with a significantly enhanced ability of these human mast cells to secrete histamine, PGD2, and leukotriene C4 upon subsequent passive sensitization with IgE and challenge with anti-IgE; 2) preincubation with IL-4 enhanced IgE-dependent mediator secretion in these cells even in the absence of significant effects on Fc epsilon RI surface expression; 3) when used together with IgE, IL-4 enhanced IgE-dependent mediator secretion in human mast cells to levels greater than those observed in cells that had been preincubated with IgE alone; and 4) batches of human mast cells generated in vitro from umbilical cord blood cells derived from different donors exhibited differences in the magnitude and pattern of histamine and lipid mediator release in response to anti-IgE challenge, both under baseline conditions and after preincubation with IgE and/or IL-4.

    View details for Web of Science ID 000079892600059

    View details for PubMedID 10228025

  • Spontaneous canine mast cell tumors express tandem duplications in the proto-oncogene c-kit EXPERIMENTAL HEMATOLOGY London, C. A., GALLI, S. J., Yuuki, T., Hu, Z. Q., Helfand, S. C., Geissler, E. N. 1999; 27 (4): 689-697

    Abstract

    Spontaneous mast cell tumors (MCT) are the most common malignant neoplasm in the dog, representing between 7% and 21% of all canine tumors, an incidence much higher than that found in humans. These tumors often behave in an aggressive manner, metastasizing to local lymph nodes, liver, spleen, and bone marrow. The proto-oncogene c-kit is known to play a critical role in the development and function of mast cells. Point mutations in the kinase domain of c-kit leading to tyrosine phosphorylation in the absence of ligand binding have been identified in three mastocytoma lines, (P815, RBL, and HMC-1), and some human patients with various forms of mastocytosis. We now demonstrate that although c-kit derived from canine MCT did not contain the previously described activating point mutations, 5 of the 11 tumors analyzed possessed novel mutations consisting of tandem duplications involving exons 11 and 12. We also show that one such duplication, detected in a canine mastocytoma cell line, was associated with constitutive phosphorylation of c-kit protein (KIT), suggesting that these mutations may contribute to the development or progression of canine MCT.

    View details for Web of Science ID 000079690600013

    View details for PubMedID 10210327

  • Mast cells as sentinels of innate immunity CURRENT OPINION IN IMMUNOLOGY Galli, S. J., Maurer, M., Lantz, C. S. 1999; 11 (1): 53-59

    Abstract

    Mast cells are widely regarded as important effector cells in immune responses associated with Th2 cells and IgE. Recent work shows that they can also contribute significantly to the expression of innate immunity; furthermore, survival in a model of acute bacterial infection that is dependent on complement and mast cells can be greatly enhanced by long-term treatment of mice with the kit ligand (stem cell factor) at least in part because of the effects of such treatment on mast cell numbers and/or function. These findings not only indicate that mast cells can represent a critical component of host defense in natural immunity but also suggest that mast cell function in this setting can be manipulated for therapeutic ends.

    View details for Web of Science ID 000078614200009

    View details for PubMedID 10047539

  • The c-kit ligand, stem cell factor, can enhance innate immunity through effects on mast cells JOURNAL OF EXPERIMENTAL MEDICINE Maurer, M., Echtenacher, B., Hultner, L., Kollias, G., Mannel, D. N., Langley, K. E., Galli, S. J. 1998; 188 (12): 2343-2348

    Abstract

    Mast cells are thought to contribute significantly to the pathology and mortality associated with anaphylaxis and other allergic disorders. However, studies using genetically mast cell-deficient WBB6F1-KitW/KitW-v and congenic wild-type (WBB6F1-+/+) mice indicate that mast cells can also promote health, by participating in natural immune responses to bacterial infection. We previously reported that repetitive administration of the c-kit ligand, stem cell factor (SCF), can increase mast cell numbers in normal mice in vivo. In vitro studies have indicated that SCF can also modulate mast cell effector function. We now report that treatment with SCF can significantly improve the survival of normal C57BL/6 mice in a model of acute bacterial peritonitis, cecal ligation and puncture (CLP). Experiments in mast cell-reconstituted WBB6F1-KitW/KitW-v mice indicate that this effect of SCF treatment reflects, at least in part, the actions of SCF on mast cells. Repetitive administration of SCF also can enhance survival in mice that genetically lack tumor necrosis factor (TNF)-alpha, demonstrating that the ability of SCF treatment to improve survival after CLP does not solely reflect effects of SCF on mast cell- dependent (or -independent) production of TNF-alpha. These findings identify c-kit and mast cells as potential therapeutic targets for enhancing innate immune responses.

    View details for Web of Science ID 000077713600015

    View details for PubMedID 9858520

  • Mast cells can secrete vascular permeability factor vascular endothelial cell growth factor and exhibit enhanced release after immunoglobulin E-dependent upregulation of Fc epsilon receptor I expression JOURNAL OF EXPERIMENTAL MEDICINE Boesiger, J., Tsai, M., Maurer, M., Yamaguchi, M., BROWN, L. F., CLAFFEY, K. P., Dvorak, H. F., Galli, S. J. 1998; 188 (6): 1135-1145

    Abstract

    Vascular permeability factor/vascular endothelial cell growth factor (VPF/VEGF) can both potently enhance vascular permeability and induce proliferation of vascular endothelial cells. We report here that mouse or human mast cells can produce and secrete VPF/VEGF. Mouse mast cells release VPF/VEGF upon stimulation through Fcepsilon receptor I (FcepsilonRI) or c-kit, or after challenge with the protein kinase C activator, phorbol myristate acetate, or the calcium ionophore, A23187; such mast cells can rapidly release VPF/VEGF, apparently from a preformed pool, and can then sustain release by secreting newly synthesized protein. Notably, the Fc epsilonRI-dependent secretion of VPF/VEGF by either mouse or human mast cells can be significantly increased in cells which have undergone upregulation of Fc epsilonRI surface expression by a 4-d preincubation with immunoglobulin E. These findings establish that at least one cell type, the mast cell, can be stimulated to secrete VPF/VEGF upon immunologically specific activation via a member of the multichain immune recognition receptor family. Our observations also identify a new mechanism by which mast cells can contribute to enhanced vascular permeability and/or angiogenesis, in both allergic diseases and other settings.

    View details for Web of Science ID 000076112700014

    View details for PubMedID 9743532

  • Involvement of Bruton's tyrosine kinase in Fc epsilon RI-dependent mast cell degranulation and cytokine production JOURNAL OF EXPERIMENTAL MEDICINE Hata, D., Kawakami, Y., Inagaki, N., Lantz, C. S., Kitamura, T., Khan, W. N., Maeda-Yamamoto, M., Miura, T., Han, W., Hartman, S. E., Yao, L., Nagai, H., Goldfeld, A. E., Alt, F. W., GALLI, S. J., Witte, O. N., Kawakami, T. 1998; 187 (8): 1235-1247

    Abstract

    We investigated the role of Bruton's tyrosine kinase (Btk) in FcepsilonRI-dependent activation of mouse mast cells, using xid and btk null mutant mice. Unlike B cell development, mast cell development is apparently normal in these btk mutant mice. However, mast cells derived from these mice exhibited significant abnormalities in FcepsilonRI-dependent function. xid mice primed with anti-dinitrophenyl monoclonal IgE antibody exhibited mildly diminished early-phase and severely blunted late-phase anaphylactic reactions in response to antigen challenge in vivo. Consistent with this finding, cultured mast cells derived from the bone marrow cells of xid or btk null mice exhibited mild impairments in degranulation, and more profound defects in the production of several cytokines, upon FcepsilonRI cross-linking. Moreover, the transcriptional activities of these cytokine genes were severely reduced in FcepsilonRI-stimulated btk mutant mast cells. The specificity of these effects of btk mutations was confirmed by the improvement in the ability of btk mutant mast cells to degranulate and to secrete cytokines after the retroviral transfer of wild-type btk cDNA, but not of vector or kinase-dead btk cDNA. Retroviral transfer of Emt (= Itk/Tsk), Btk's closest relative, also partially improved the ability of btk mutant mast cells to secrete mediators. Taken together, these results demonstrate an important role for Btk in the full expression of FcepsilonRI signal transduction in mast cells.

    View details for Web of Science ID 000073287900008

    View details for PubMedID 9547335

  • P- and E-selectins are required for the leukocyte recruitment, but not the tissue swelling, associated with IgE- and mast cell-dependent inflammation in mouse skin LABORATORY INVESTIGATION de Mora, F., Williams, C. M., Frenette, P. S., Wagner, D. D., HYNES, R. O., GALLI, S. J. 1998; 78 (4): 497-505

    Abstract

    Many studies, in both experimental animal and human systems, have indicated that P- and/or E-selectins may contribute importantly to the leukocyte recruitment that occurs in association with mast cell-dependent inflammatory responses. We used mice that genetically lack P-selectin (P -/-), E-selectin (E -/-), or both selectins (P/E -/-) to investigate the possible roles of these selectins in the IgE- and mast cell-dependent recruitment of neutrophils to the skin of mice. We found that a lack of either or both selectins had little or no effect on the extent of mast cell degranulation or the tissue swelling associated with these reactions. Moreover, a lack of either P- or E-selectin alone did not reduce the neutrophil infiltration at the reaction sites. However, mice lacking both P- and E-selectins exhibited an almost complete ablation of IgE- and mast cell-dependent neutrophil recruitment. These findings show that P- and E-selectins can express overlapping functions in leukocyte recruitment associated with IgE- and mast cell-dependent cutaneous late-phase reactions in mouse skin, and that a lack of both selectins results in a virtual elimination of IgE-dependent leukocyte recruitment.

    View details for Web of Science ID 000073169200013

    View details for PubMedID 9564894

  • Role for interleukin-3 in mast-cell and basophil development and in immunity to parasites NATURE Lantz, C. S., Boesiger, J., Song, C. H., MACH, N., Kobayashi, T., Mulligan, R. C., Nawa, Y., Dranoff, G., GALLI, S. J. 1998; 392 (6671): 90-93

    Abstract

    The cytokine interleukin-3 (IL-3), which can be derived from T cells and other sources, is a potentially important link between the immune and haematopoietic systems. IL-3 may be particularly critical for the development, survival and function of tissue mast cells and blood basophils, which are thought to be important effector cells in immunity to parasites and other immunological responses, such as allergic reactions. Here we show, using IL-3-deficient mice, that IL-3 is not essential for the generation of mast cells or basophils under physiological conditions, but that it does contribute to increased numbers of tissue mast cells, enhanced basophil production, and immunity in mice infected with the nematode Stronglyoides venezuelensis. Parasite expulsion and mast-cell development are impaired even more severely in IL-3-deficient mice that also show a marked reduction in signalling by c-kit. These findings establish a role for IL-3 in immunity to parasites and indicate that one of the functions of IL-3 in host defence against infection is to expand populations of haematopoietic effector cells.

    View details for Web of Science ID 000072373000056

    View details for PubMedID 9510253

  • Involvement of interleukin-3 in delayed-type hypersensitivity BLOOD MACH, N., Lantz, C. S., GALLI, S. J., Reznikoff, G., Mihm, M., Small, C., Granstein, R., Beissert, S., Sadelain, M., Mulligan, R. C., Dranoff, G. 1998; 91 (3): 778-783

    Abstract

    The in vivo functions of interleukin-3 (IL-3) were investigated by generating IL-3-deficient mice. Although hematopoiesis was unimpaired in homozygous mutant animals, contact hypersensitivity reactions were compromised. IL-3 was required for efficient priming of hapten-specific contact hypersensitivity responses, but was dispensable for T-cell-dependent sensitization to tumor cells. These findings reveal a critical role for IL-3 in some forms of delayed-type hypersensitivity.

    View details for Web of Science ID 000071644000007

    View details for PubMedID 9446636

  • Impaired mast cell-dependent natural immunity in complement C3-deficient mice NATURE Prodeus, A. P., Zhou, X. N., Maurer, M., GALLI, S. J., Carroll, M. C. 1997; 390 (6656): 172-175

    Abstract

    The complement system is widely regarded as essential for normal inflammation, not least because of its ability to activate mast cells. However, recent studies have called into question the importance of complement in several examples of mast cell-dependent inflammatory responses. To investigate the role of complement in mast cell-dependent natural immunity, we examined the responses of complement-deficient mice to caecal ligation and puncture, a model of acute septic peritonitis that is dependent on mast cells and tumour necrosis factor-alpha (TNF-alpha). We found that C4- or C3-deficient mice were much more sensitive to caecal ligation and puncture than wild-type (WT) controls (100% versus 20% in 24-h mortality, respectively). C3-deficient mice also exhibited reductions in peritoneal mast cell degranulation, production of TNF-alpha, neutrophil infiltration and clearance of bacteria. Treating the C3-deficient mice with purified C3 protein enhanced activation of peritoneal mast cells, TNF-alpha production, neutrophil recruitment, opsonophagocytosis of bacteria and resistance to caecal ligation and puncture, confirming that the defects were complement-dependent. These results provide formal evidence that complement activation is essential for the full expression of innate immunity in this mast cell-dependent model of bacterial infection.

    View details for Web of Science ID A1997YF49400054

    View details for PubMedID 9367154

  • Expression of functional TrkA receptor tyrosine kinase in the HMC-1 human mast cell line and in human mast cells BLOOD Tam, S. Y., Tsai, M., Yamaguchi, M., Yano, K., Butterfield, J. H., GALLI, S. J. 1997; 90 (5): 1807-1820

    Abstract

    Nerve growth factor (NGF) can influence mast cell development and function in murine rodents by interacting with its receptors on mast cells. We now report the identification of mRNA transcripts of full-length tyrosine kinase-containing trkA, trkB, and trkC neurotrophin receptor genes in HMC-1 human mast cell leukemia cells. Although HMC-1 cells lacked p75 mRNA, they expressed transcripts for the exon-lacking splice variant of trkA (trkAI), truncated trkB (trkB.T1), and truncated trkC. By flow cytometry, HMC-1 cells exhibited expression of TrkA, TrkB, and TrkC receptor proteins containing full-length tyrosine kinase domains. NGF stimulation of HMC-1 cells induced tyrosine phosphorylation of TrkA protein, increased expression of the early response genes c-fos and NGF1-A, and activation of ERK-mitogen-activated protein (MAP) kinase, results which indicate that TrkA receptors in HMC-1 cells are fully functional. Highly purified populations of human lung mast cells expressed mRNAs for trkA, trkB and trkC, whereas preparations of human umbilical cord blood-derived mast cells expressed mRNAs for trkA and trkC, but not trkB. Moreover, preparations of human umbilical cord blood-derived immature mast cells not only expressed mRNA transcript and protein for TrkA, but exhibited significantly higher numbers of chymase-positive cells after the addition of NGF to their culture medium for 3 weeks. In addition, HMC-1 cells expressed mRNAs for NGF, brain-derived neurotrophic factor (BDNF), and neurotrophin-3 (NT-3), the cognate ligands for TrkA, TrkB, and TrkC, whereas NGF and BDNF transcripts were detectable in human umbilical cord blood mast cell preparations. Taken together, our findings show that human mast cells express a functional TrkA receptor tyrosine kinase and indicate that NGF may be able to promote certain aspects of mast cell development and/or maturation in humans. Our studies also raise the possibility that human mast cells may represent a potential source for neurotrophins.

    View details for Web of Science ID A1997XX16600009

    View details for PubMedID 9292513

  • Complexity and redundancy in the pathogenesis of asthma: Reassessing the roles of mast cells and T cells JOURNAL OF EXPERIMENTAL MEDICINE GALLI, S. J. 1997; 186 (3): 343-347

    View details for Web of Science ID A1997XP99800001

    View details for PubMedID 9265074

  • The Paul Kallos Memorial Lecture. The mast cell: a versatile effector cell for a challenging world. International archives of allergy and immunology GALLI, S. J. 1997; 113 (1-3): 14-22

    Abstract

    Mast cells are phenotypically and functionally versatile effector cells. When activated by IgE-dependent or other mechanisms, mast cells can produce a diverse array of mediators including TNF-alpha and many other cytokines. Moreover, mast cells can express increased numbers of high-affinity surface receptors for IgE (Fc epsilonRI) and enhanced levels of IgE-dependent mediator secretion in response to elevations in concentrations of IgE. These characteristics (and others) have suggested diverse potential roles for mast cells in health and disease. To test specific hypotheses about mast cell function in allergic reactions and other biological responses in vivo, one can employ genetically mast-cell-deficient Kit(W)/Kit(W-v) mice which do or do not contain adoptively transferred mast cell populations derived from genetically compatible wild-type mice or mice with mutations that influence mast cell biology. Such work has already indicated that mast cells (and, in some cases, mast-cell-derived cytokines) can have a critical role in the expression of the acute, late-phase and chronic components of IgE-dependent allergic inflammation and can influence the development of an important functional consequence of such reactions: airways hyperresponsiveness. However, mast cells can also perform important beneficial roles in host defense, both in IgE-dependent immune responses to certain parasites and in natural immunity to bacterial infection.

    View details for PubMedID 9130474

  • Differential release of mast cell interleukin-6 via c-kit BLOOD Gagari, E., Tsai, M., Lantz, C. S., FOX, L. G., GALLI, S. J. 1997; 89 (8): 2654-2663

    Abstract

    Mast cells represent a potential source of interleukin-6 (IL-6) and other cytokines that have been implicated in host defense, tissue maintenance/remodeling, immunoregulation, and many other biologic responses. In acquired immune responses to parasites or allergens, the extensive IgE-dependent activation of mast cells via Fc epsilonRI can result in the release of large quantities of biogenic amines that are stored in the cells' cytoplasmic granules as well as the production of lipid mediators and many cytokines; these products together can orchestrate an intense inflammatory response. We now report that activation of mouse mast cells via c-kit, the receptor for the pleiotropic survival/growth factor, stem cell factor (SCF), can induce the release of IL-6. Upon challenge with SCF, bone marrow-derived cultured mouse mast cells (BMCMCs) released amounts of IL-6 that were greater than 100-fold more than those produced by unstimulated cells, but that were substantially less than those produced in response to IgE and specific antigen. Moreover, BMCMCs released IL-6 upon challenge with concentrations of SCF that resulted in little or no detectable release of tumor necrosis factor-alpha, leukotriene C4, histamine, or serotonin. These findings indicate that SCF, a widely expressed protein that is critical for mast cell development and survival, can also regulate the differential release of mast cell mediators.

    View details for Web of Science ID A1997WV14600004

    View details for PubMedID 9108382

  • IgE regulates mouse basophil Fc epsilon RI expression in vivo JOURNAL OF IMMUNOLOGY Lantz, C. S., Yamaguchi, M., Oettgen, H. C., Katona, I. M., Miyajima, I., Kinet, J. P., GALLI, S. J. 1997; 158 (6): 2517-2521

    Abstract

    The binding of IgE to high-affinity IgE receptors (Fc epsilon RI) on the surface of mast cells and basophils primes these cells to secrete a panel of proinflammatory mediators upon subsequent exposure to specific Ag. We now find that the level of Fc epsilon RI expression on bone marrow basophils in mice infected with the nematode Strongyloides venezuelensis exhibits a strong positive correlation with the serum concentration of IgE, as was previously reported for human blood basophils. Moreover, the administration of IgE in vivo can significantly upregulate Fc epsilon RI expression on mouse basophils, and genetically IgE-deficient (IgE -/-) mice exhibit a dramatic (approximately 81%) reduction of basophil Fc epsilon RI expression compared with the corresponding normal (IgE +/+) mice. The finding that IgE can be a major regulator of mouse basophil Fc epsilon RI expression in vivo identifies a potentially important mechanism for enhancing the expression of effector cell function in IgE-dependent allergic reactions or immunologic responses to parasites.

    View details for Web of Science ID A1997WM43500003

    View details for PubMedID 9058781

  • Absence of Fc(epsilon)RI alpha chain results in upregulation of Fc gamma RIII-dependent mast cell degranulation and anaphylaxis - Evidence of competition between Fc(epsilon)RI and Fc gamma RIII for limiting amounts of FcR beta and gamma chains JOURNAL OF CLINICAL INVESTIGATION Dombrowicz, D., Flamand, V., Miyajima, I., Ravetch, J. V., GALLI, S. J., Kinet, J. P. 1997; 99 (5): 915-925

    Abstract

    In mouse mast cells, both Fc epsilonRI and Fc gammaRIII are alpha beta gamma2 tetrameric complexes in which different alpha chains confer IgE or IgG ligand recognition while the signaling FcR beta and gamma chains are identical. We used primarily noninvasive techniques (changes in body temperature, dye extravasation) to assess systemic anaphylactic responses in nonanesthetized wild-type, Fc epsilonRI alpha chain -/- and FcR gamma chain -/- mice. We confirm that systemic anaphylaxis in mice can be mediated largely through IgG1 and Fc gammaRIII and we provide direct evidence that these responses reflect activation of Fc gammaRIII rather than Fc gammaRI. Furthermore, we show that Fc gammaRIII-dependent responses are more intense in normal than in congenic mast cell-deficient KitW/KitW-v mice, indicating that Fc gammaRIII responses have mast cell-dependent and -independent components. Finally, we demonstrate that the upregulation of cell surface expression of Fc gammaRIII seen in Fc epsilonRI alpha chain -/- mice corresponds to an increased association of Fc gammaRIII alpha chains with FcR beta and gamma chains and is associated with enhanced Fc gammaRIII-dependent mast cell degranulation and systemic anaphylactic responses. Therefore, the phenotype of the Fc epsilonRI alpha chain -/- mice suggests that expression of Fc epsilonRI and Fc gammaRIII is limited by availability of the FcR beta and gamma chains and that, in normal mice, changes in the expression of one receptor (Fc epsilonRI) may influence the expression of functional responses dependent on the other (Fc gammaRIII).

    View details for Web of Science ID A1997WN15200015

    View details for PubMedID 9062349

  • Systemic anaphylaxis in the mouse can be mediated largely through IgG, and Fc gamma RIII - Assessment of the cardiopulmonary changes, mast cell degranulation, and death associated with active or IgE- or IgG(1)-dependent passive anaphylaxis JOURNAL OF CLINICAL INVESTIGATION Miyajima, I., Dombrowicz, D., Martin, T. R., Ravetch, J. V., Kinet, J. P., GALLI, S. J. 1997; 99 (5): 901-914

    Abstract

    We attempted to elicit active anaphylaxis to ovalbumin, or passive IgE- or IgG1-dependent anaphylaxis, in mice lacking either the Fc epsilonRI alpha chain or the FcR gamma chain common to Fc epsilonRI and Fc gammaRI/III, or in mice lacking mast cells (KitW/ KitW-v mice), and compared the responses to those in the corresponding wild-type mice. We found that the FcR gamma chain is required for the death, as well as for most of the pathophysiological changes, associated with active anaphylaxis or IgE- or IgG1-dependent passive anaphylaxis. Moreover, some of the physiological changes associated with either active, or IgG1-dependent passive, anaphylactic responses were significantly greater in Fc epsilonRI alpha chain -/- mice than in the corresponding normal mice. Finally, while both KitW/KitW-v and congenic +/+ mice exhibited fatal active anaphylaxis, mast cell-deficient mice exhibited weaker physiological responses than the corresponding wild-type mice in both active and IgG1-dependent passive systemic anaphylaxis. Our findings strongly suggest that while IgE antibodies and Fc epsilonRI may influence the intensity and/or kinetics of some of the pathophysiological changes associated with active anaphylaxis in the mouse, the mortality associated with this response can be mediated largely by IgG1 antibodies and Fc gammaRIII.

    View details for Web of Science ID A1997WN15200014

    View details for PubMedID 9062348

  • IgE enhances mouse mast cell Fc epsilon RI expression in vitro and in vivo: Evidence for a novel amplification mechanism in IgE-dependent reactions JOURNAL OF EXPERIMENTAL MEDICINE Yamaguchi, M., Lantz, C. S., Oettgen, H. C., Katona, I. M., Fleming, T., Miyajima, I., Kinet, J. P., GALLI, S. J. 1997; 185 (4): 663-672

    Abstract

    The binding of immunoglobulin E (IgE) to high affinity IgE receptors (Fc(epsilon)RI) expressed on the surface of mast cells primes these cells to secrete, upon subsequent exposure to specific antigen, a panel of proinflammatory mediators, which includes cytokines that can also have immunoregulatory activities. This IgE- and antigen-specific mast cell activation and mediator production is thought to be critical to the pathogenesis of allergic disorders, such as anaphylaxis and asthma, and also contributes to host defense against parasites. We now report that exposure to IgE results in a striking (up to 32-fold) upregulation of surface expression of Fc(epsilon)RI on mouse mast cells in vitro or in vivo. Moreover, baseline levels of Fc(epsilon)RI expression on peritoneal mast cells from genetically IgE-deficient (IgE -/-) mice are dramatically reduced (by approximately 83%) compared with those on cells from the corresponding normal mice. In vitro studies indicate that the IgE-dependent upregulation of mouse mast cell Fc(epsilon)RI expression has two components: an early cycloheximide-insensitive phase, followed by a later and more sustained component that is highly sensitive to inhibition by cycloheximide. In turn, IgE-dependent upregulation of Fc(epsilon)RI expression significantly enhances the ability of mouse mast cells to release serotonin, interleukin-6 (IL-6), and IL-4 in response to challenge with IgE and specific antigen. The demonstration that IgE-dependent enhancement of mast cell Fc(epsilon)RI expression permits mast cells to respond to antigen challenge with increased production of proinflammatory and immunoregulatory mediators provides new insights into both the pathogenesis of allergic diseases and the regulation of protective host responses to parasites.

    View details for Web of Science ID A1997WK03800008

    View details for PubMedID 9034145

  • Earthquakes: Slow down for safety NATURE Segall, P. 1996; 383 (6595): 21-22

    View details for Web of Science ID A1996VF29500027

    View details for PubMedID 8609979

  • Genetic analysis reveals cell type-specific regulation of receptor tyrosine kinase c-Kit by the protein tyrosine phosphatase SHP1 JOURNAL OF EXPERIMENTAL MEDICINE Lorenz, U., Bergemann, A. D., Steinberg, H. N., Flanagan, J. G., Li, X. T., GALLI, S. J., Neel, B. G. 1996; 184 (3): 1111-1126

    Abstract

    Receptor protein tyrosine kinases (RTKs) transmit downstream signals via interactions with secondary signaling molecules containing SH2 domains. Although many SH2-phosphotyrosyl interactions have been defined in vitro, little is known about the physiological significance of specific RTK/SH2 interactions in vivo. Also, little is known about the mechanisms by which specific RTKs interact with and/or are regulated by specific protein tyrosine phosphatases (PTPs). To address such issue, we carried out a genetic analysis of the previously reported biochemical interaction between the RTK c-Kit, encoded at the W locus, and the SH2-containing non-transmembrane PTP SHP1, encoded at the motheaten (me) locus (1). Mice carrying a kinase-defective allele of c-Kit (Wv/+) were crossed with me/+ mice, which carry one effectively null allele of SHP1, and then backcrossed to generate all possible allelic combinations. Our results indicate strong intergenic complementation between these loci in hematopoietic progenitor cells. Compared to progenitors purified from normal mice, bone marrow progenitor cells (lin-) from me/me mice markedly hyper-proliferated in response to Kit ligand (KL). stimulation. Superimposition of the me/me genotype increased the number of one marrow-derived CFU-E from Wv/+ mice. Conversely, the presence of one or two copies of Wv decreased the number of macrophages and granulocytes in me/me lung, skin, peripheral blood and bone marrow, thereby decreasing the severity of the me/me phenotype. The decrease in dermal mast cells in Wv/Wv mice was rescued to levels found in Wv/+mice by superimposition of the me/me genotype. Surprisingly, however, the presence or absence of SHP1 had no effect on the proliferative response of bone marrow-derived cultured mast cells to KL or IL3 ex vivo. Nevertheless, the immediate-early response to KL stimulation, as measured by KL-induced tyrosyl phosphorylation, was substantially increased in mast cells from Wv/+:me/me compared to Wv/ +:+/+ mice, strongly suggesting that SHP1 directly dephosphorylates and regulates c-Kit. Taken together, our results establish that SHP1 negatively regulates signaling from c-Kit in vivo, but in a cell type-specific manner.

    View details for Web of Science ID A1996VG86800032

    View details for PubMedID 9064328

  • Recombinant human stem cell factor (kit ligand) promotes human mast cell and melanocyte hyperplasia and functional activation in vivo JOURNAL OF EXPERIMENTAL MEDICINE Costa, J. J., Demetri, G. D., Harrist, T. J., Dvorak, A. M., Hayes, D. F., Merica, E. A., Menchaca, D. M., Gringeri, A. J., SCHWARTZ, L. B., GALLI, S. J. 1996; 183 (6): 2681-2686

    Abstract

    Stem cell factor (SCF), also known as mast cell growth factor, kit ligand, and steel factor, is the ligand for the tyrosine kinase receptor (SCFR) that is encoded by the c-kit proto-oncogene. We analyzed the effects of recombinant human SCF (r-hSCF, 5-50 micrograms/kg/day, injected subcutaneously) on mast cells and melanocytes in a phase I study of 10 patients with advanced breast carcinoma. A wheal and flare reaction developed at each r-hSCF injection site; by electron microscopy, most dermal mast cells at these sites exhibited extensive, anaphylactic-type degranulation. A 14-d course of r-hSCF significantly increased dermal mast cell density at sites distant to those injected with the cytokine and also increased both urinary levels of the major histamine metabolite, methyl-histamine, and serum levels of mast cell alpha-tryptase. Five subjects developed areas of persistent hyperpigmentation at r-hSCF injection sites; by light microscopy, these sites exhibited markedly increased epidermal melanization and increased numbers of melanocytes. The demonstration that r-hSCF can promote both the hyperplasia and the functional activation of human mast cells and melanocytes in vivo has implications for our understanding of the role of endogenous SCF in health and disease. These findings also indicate that the interaction between SCF and its receptor represents a potential therapeutic target for regulating the numbers and functional activity of both mast cells and cutaneous melanocytes.

    View details for Web of Science ID A1996UT40100031

    View details for PubMedID 8676090

  • Identification of a committed precursor for the mast cell lineage SCIENCE Rodewald, H. R., Dessing, M., Dvorak, A. M., GALLI, S. J. 1996; 271 (5250): 818-822

    Abstract

    Mast cells originate from hematopoietic stem cells, but the mast cell-committed precursor has not been identified. In the study presented here, a cell population in murine fetal blood that fulfills the criteria of progenitor mastocytes was identified. It is defined by the phenotype Thy-1loc-Kithi, contains cytoplasmic granules, and expresses RNAs encoding mast cell-associated proteases but lacks expression of the high-affinity immunoglobulin E receptor. Thy-1loc-Kithi cells generated functionally competent mast cells at high frequencies in vitro but lacked developmental potential for other hematopoietic lineages. When transferred intraperitoneally, this population reconstituted the peritoneal mast cell compartment of genetically mast cell-deficient W/Wv mice to wild-type levels.

    View details for Web of Science ID A1996TU69400050

    View details for PubMedID 8629001

  • STEM-CELL FACTOR CONTRIBUTES TO INTESTINAL MUCOSAL MAST-CELL HYPERPLASIA IN RATS INFECTED WITH NIPPOSTRONGYLUS-BRASILIENSIS OR TRICHINELLA-SPIRALIS, BUT ANTI-STEM CELL FACTOR TREATMENT DECREASES PARASITE EGG-PRODUCTION DURING N-BRASILIENSIS INFECTION BLOOD Newlands, G. F., Miller, H. R., MacKellar, A., GALLI, S. J. 1995; 86 (5): 1968-1976

    Abstract

    We assessed the effects of the c-kit ligand, stem cell factor (SCF), in the jejunal mucosal mast cell hyperplasia that occurs during infection with the intestinal nematodes, Nippostrongylus brasiliensis or Trichinella spiralis in rats. Compared with vehicle-treated rats, rats treated with SCF (25 micrograms/kg/d, intravenous [i.v.] for 14 days) during N brasiliensis infection exhibited significantly higher levels of the rat mucosal mast cell (MMC)-associated protease, rat mast cell protease II (RMCP II) in the jejunum and serum on day 8 of infection, but not on days 10 or 15 of infection. By contrast, in comparison to rats treated with normal sheep IgG, rats treated with a polyclonal sheep antirat SCF antibody exhibited markedly decreased numbers of jejunal MMCs, levels of jejunal RMCP II, and serum concentrations of RMCP II during infection with either nematode, particularly at the earlier intervals of infection (< or = day 10). Taken together, these findings indicate that SCF importantly contributes to MMC hyperplasia and/or survival during N brasiliensis or T spiralis infection in rats, but that levels of endogenous SCF are adequate to sustain near maximal MMC hyperplasia during infection with these nematodes. Notably, treatment of rats with SCF somewhat increased, and treatment with anti-SCF significantly decreased, parasite egg production during N brasiliensis infection. This finding raises the interesting possibility that certain activities of intestinal MMCs may contribute to parasite fecundity during infection with this nematode.

    View details for Web of Science ID A1995RT38600039

    View details for PubMedID 7544650

  • DEXAMETHASONE OR CYCLOSPORINE-A SUPPRESS MAST CELL-LEUKOCYTE CYTOKINE CASCADES - MULTIPLE MECHANISMS OF INHIBITION OF IGE-DEPENDENT AND MAST-CELL-DEPENDENT CUTANEOUS INFLAMMATION IN THE MOUSE JOURNAL OF IMMUNOLOGY Wershil, B. K., Furuta, G. T., Lavigne, J. A., Choudhury, A. R., Wang, Z. S., GALLI, S. J. 1995; 154 (3): 1391-1398

    Abstract

    In allergic diseases, exposure of sensitized subjects to allergen induces the activation of tissue mast cells that results in an immediate-type hypersensitivity response and, in some individuals, a a late phase response. We previously have reported that the neutrophil infiltration associated with IgE-dependent cutaneous inflammation in mice is mast cell-dependent and that TNF-alpha contributes significantly to this response. We report here that either dexamethasone or cyclosporin A can inhibit mouse mast cell TNF-alpha production in vitro, and that these agents also can significantly suppress the tissue swelling and leukocyte infiltration associated with two forms of TNF-alpha-associated inflammation in vivo: the entirely IgE- and mast cell-dependent inflammation at sites of passive cutaneous anaphylaxis reactions and the entirely TNF-alpha-dependent inflammation that is elicited by the direct intradermal injection of recombinant mouse TNF-alpha. Taken together, our in vitro and in vivo findings in mice indicate that dexamethasone or cyclosporin A can have at least three actions that interfere with the pathogenesis of IgE, mast cell, and cytokine-dependent inflammatory reactions:suppression of the IgE-dependent increase in TNF-alpha mRNA by mast cells, inhibition of the IgE-dependent production of TNF-alpha protein by mast cells, and diminution of the responsiveness of target cells to TNF-alpha. Our findings in mice raise the possibility that similar actions of these agents in humans may account for some of the clinical efficacy of corticosteroids and cyclosporin A in allergic diseases.

    View details for Web of Science ID A1995QC54600045

    View details for PubMedID 7822805

  • PROMOTION OF MOUSE FIBROBLAST COLLAGEN GENE-EXPRESSION BY MAST-CELLS STIMULATED VIA THE FC(EPSILON)RI - ROLE FOR MAST CELL-DERIVED TRANSFORMING GROWTH-FACTOR-BETA AND TUMOR-NECROSIS-FACTOR-ALPHA JOURNAL OF EXPERIMENTAL MEDICINE Gordon, J. R., GALLI, S. J. 1994; 180 (6): 2027-2037

    Abstract

    Chronic allergic diseases and other disorders associated with mast cell activation can also be associated with tissue fibrosis, but a direct link between mast cell mediator release and fibroblast collagen gene expression has not been established. Using in situ hybridization, we show that the elicitation of an IgE-dependent passive cutaneous anaphylaxis (PCA) reaction in mice results in a transient, but marked augmentation of steady state levels of type alpha-1 (I) collagen mRNA in the dermis. While peak levels of collagen mRNA expression in the skin are observed 16-24 h after mast cell activation, substantial numbers of dermal cells are strongly positive for collagen mRNA at 1 and 2 h after antigen challenge, before circulating inflammatory cells are recruited into the tissues. Furthermore, experiments in mast cell-reconstituted or genetically mast cell-deficient WBB6F1-W/Wv mice demonstrate that the increased expression of collagen mRNA at sites of PCA reactions is entirely mast cell dependent. In vitro studies show that the supernatants of mouse serosal mast cells activated via the Fc epsilon RI markedly increase type alpha-1 (I) collagen mRNA levels in mouse embryonic skin fibroblasts, and also upregulate collagen secretion by these cells. The ability of mast cell supernatants to induce increased steady state levels of collagen mRNA in mouse skin fibroblasts is markedly diminished by absorption with antibodies specific for either of two mast cell-derived cytokines, transforming growth factor beta (TGF-beta 1) or tumor necrosis factor alpha (TNF-alpha), and is eliminated entirely by absorption with antibodies against both cytokines. Taken together, these findings demonstrate that IgE-dependent mouse mast cell activation can induce a transient and marked increase in steady state levels of type alpha-1 (I) collagen mRNA in dermal fibroblasts and that mast cell-derived TGF-beta 1 and TNF-alpha importantly contribute to this effect.

    View details for Web of Science ID A1994PU36100003

    View details for PubMedID 7964480

  • CYTOPLASMIC GRANULE FORMATION IN MOUSE PANCREATIC ACINAR-CELLS - EVIDENCE FOR FORMATION OF IMMATURE GRANULES (CONDENSING VACUOLES) BY AGGREGATION AND FUSION OF PROGRANULES OF UNIT SIZE, AND FOR REDUCTIONS IN MEMBRANE-SURFACE AREA AND IMMATURE GRANULE VOLUME DURING GRANULE MATURATION CELL AND TISSUE RESEARCH LEW, S., Hammel, I., GALLI, S. J. 1994; 278 (2): 327-336

    Abstract

    We used a computer-assisted morphometry approach to analyze quantitatively the process of cytoplasmic granule formation in mouse pancreatic acinar cells stimulated with pilocarpine to induce secretion. Our findings suggest that each condensing vacuole/immature granule of pancreatic acinar cells is formed by the progressive aggregation of 106 to 128 "unit progranules" of narrowly fixed volume, define a range of 7.7 to 9.2 for the factor of volume condensation between the largest immature granules and the mature unit granule, and predict that the formation of a single mature unit granule by the aggregation and fusion of unit progranules involves a net reduction of at least 95% in the amount of membrane surface area associated with these structures.

    View details for Web of Science ID A1994PM03400016

    View details for PubMedID 8001087

  • PIECEMEAL DEGRANULATION OF MAST-CELLS IN THE INFLAMMATORY EYELID LESIONS OF INTERLEUKIN-4 TRANSGENIC MICE - EVIDENCE OF MAST-CELL HISTAMINE-RELEASE IN-VIVO BY DIAMINE OXIDASE-GOLD ENZYME-AFFINITY ULTRASTRUCTURAL CYTOCHEMISTRY BLOOD Dvorak, A. M., Tepper, R. I., Weller, P. F., Morgan, E. S., ESTRELLA, P., MONAHANEARLEY, R. A., GALLI, S. J. 1994; 83 (12): 3600-3612

    Abstract

    We used light and electron microscopy to analyze the eyelid inflammation that develops in transgenic mice that overexpress interleukin-4 (IL-4; Tepper et al, Cell 62:457, 1990). Analysis of alkaline Giemsa-stained plastic sections examined by light microscopy (Dvorak et al, J Exp Med 132:558, 1970), as well as by routine transmission electron microscopy, indicated that the mast cells in the inflammatory eyelid lesions were undergoing piecemeal degranulation, a form of secretion in which the cells' cytoplasmic granules exhibit characteristic morphologic changes that are thought to be associated with the prolonged, vesicle-mediated release of the granules' constituents. Moreover, by using a newly reported enzyme affinity-gold method, which stains histamine based on binding to diamine oxidase-gold (Dvorak et al, J Histochem Cytochem 41:787, 1993), we show that these activated mast cells had released much of their histamine content. The eyelid lesions also exhibited increased numbers of mast cells; interstitial fibrosis, particularly around cutaneous nerves and blood vessels; activated fibroblasts; focal axonal damage; venules with endothelial cells containing numerous vesiculo-vacuolar organelles; and infiltrates of neutrophils and eosinophils. Our findings illustrate that overexpression of the IL-4 gene in vivo can result in eyelid lesions associated with piecemeal degranulation of mast cells, as well as tissue fibrosis and a variety of other pathologic changes. These results also represent the first direct morphologic evidence for histamine secretion by mast cells in vivo.

    View details for Web of Science ID A1994NR90500023

    View details for PubMedID 7515717

  • MAST-CELLS AUGMENT LESION SIZE AND PERSISTENCE DURING EXPERIMENTAL LEISHMANIA-MAJOR INFECTION IN THE MOUSE JOURNAL OF IMMUNOLOGY Wershil, B. K., Theodos, C. M., GALLI, S. J., Titus, R. G. 1994; 152 (9): 4563-4571

    Abstract

    Mast cells are a source of a variety of cytokines that may influence the host response to Leishmania major. To investigate the role of mast cells during L. major infection, we performed a morphometric analysis of mast cells at cutaneous sites in resistant C57BL/6 mice and susceptible BALB/c mice injected with L. major. Extensive dermal mast cell degranulation was found at sites of L. major infection in both strains of mice. We also examined the course of L. major infection in genetically mast cell-deficient WBB6F1-W/Wv or WCB6F1-Sl/Sld mice, their respective congenic normal (WBB6F1-(+/+) or WCB6F1-(+/+)) littermates, and WBB6F1-W/Wv mice that had been selectively and locally repaired of their cutaneous mast cell deficiency. We found that mast cells significantly augmented the intensity and maximal size of the cutaneous lesions at sites of L. major infection, and in some cases substantially prolonged the persistence of the reactions. However, the lesions ultimately resolved in both the mast cell-deficient and the congenic normal mice. In addition, the presence or absence of mast cells had little or no effect on the numbers of viable parasites recovered from the cutaneous lesions. Moreover, mast cell-deficient W/Wv mice and the congenic normal (+/+) mice produced similar levels of IFN-gamma mRNA in lymph nodes draining the cutaneous lesions whereas no IL-4 mRNA was detectable. Taken together, these data suggest that mast cells significantly augment the size of cutaneous lesions during L. major infection in mice. However, mast cells do not appear to influence significantly either the parasite burden or the ultimate resolution of the infection.

    View details for Web of Science ID A1994NG71900036

    View details for PubMedID 8157970

  • IDENTIFICATION OF IGE-BEARING CELLS IN THE LATE-PHASE RESPONSE TO ANTIGEN IN THE LUNG AS BASOPHILS AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY Guo, C. B., Liu, M. C., GALLI, S. J., Bochner, B. S., KAGEYSOBOTKA, A., LICHTENSTEIN, L. M. 1994; 10 (4): 384-390

    Abstract

    We have carried out studies to ascertain whether the histamine-containing, IgE-bearing cells found in the bronchoalveolar lavage (BAL) fluid obtained during the late-phase response following subsegmental antigen challenge of human airways are predominantly basophils or mast cells. Four lines of evidence suggest that most are basophils: (1) The cells fulfill morphologic criteria for light microscopy. (2) Cell surface markers determined by immunofluorescence and flow cytometry revealed that the IgE-bearing cells express the leukocyte antigens Fc gamma RII and the beta 2 integrins, LFA-1 and Mac-1, but do not express the mast cell-associated c-kit receptor for stem cell factor. (3) The late-phase histamine-containing cells in late-phase BAL fluids have the functional characteristics of basophils in their secretory responses to anti-IgE, the f-met peptide, and phorbol ester TPA. (4) The cells have a functional histamine type 2 receptor, a characteristic of basophils, not mast cells. We conclude that basophils infiltrate the lower airways hours after antigen exposure. These cells may be responsible for the mediator release observed at that time.

    View details for Web of Science ID A1994NE36700006

    View details for PubMedID 7510984

  • THE KIT-LIGAND, STEM-CELL FACTOR ADVANCES IN IMMUNOLOGY, VOL 55 GALLI, S. J., Zsebo, K. M., Geissler, E. N. 1994; 55: 1-96

    View details for Web of Science ID A1994BZ73F00001

    View details for PubMedID 7508174

  • EFFECTS OF INTERLEUKIN-3 WITH OR WITHOUT THE C-KIT LIGAND, STEM-CELL FACTOR, ON THE SURVIVAL AND CYTOPLASMIC GRANULE FORMATION OF MOUSE BASOPHILS AND MAST-CELLS IN-VITRO AMERICAN JOURNAL OF PATHOLOGY Dvorak, A. M., SEDER, R. A., PAUL, W. E., Morgan, E. S., GALLI, S. J. 1994; 144 (1): 160-170

    Abstract

    We assessed the ultrastructure and the cell-surface expression of receptors for immunoglobulin E (Fc epsilon R), and c-kit, the receptor for stem cell factor (SCF), in mouse basophils and mast cells present in short-term cultures of mouse bone marrow cells in interleukin-3 (IL-3) with or without SCF. Basophils did not develop increased numbers of cytoplasmic granules and underwent apoptosis in cultures containing IL-3 and SCF, whereas mast cells thrived and developed increased numbers of granules. Basophils were nearly all Fc epsilon R+ c-kit- when sorted after culture in IL-3 and SCF; most mast cells were Fc epsilon R+ c-kit+. However, a second population of Fc epsilon R+ c-kit- mast cells was present after culture in IL-3 and SCF. These c-kit receptor-negative mast cells were less mature than c-kit+ mast cells and contained significantly fewer cytoplasmic granules than the c-kit+ mast cells present in the same cultures (P < 0.001). Thus, mouse basophils express little or no c-kit receptor on their surface, nor can they survive for long periods in SCF-supplemented cultures. By contrast, mouse mast cells seem to express the Fc epsilon R early in their development, even before they express detectable c-kit receptors on their surface. IL-3 promotes cytoplasmic granule formation in immature mast cells, but even more granules are formed when c-kit receptor-positive immature mast cells are cultured in both SCF and IL-3.

    View details for Web of Science ID A1994MR31000017

    View details for PubMedID 7507298

  • EFFECTS OF CHRONIC TREATMENT WITH THE C-KIT LIGAND, STEM-CELL FACTOR, ON IMMUNOGLOBULIN-E DEPENDENT ANAPHYLAXIS IN MICE - GENETICALLY MAST-CELL DEFICIENT SL/SL(D) MICE ACQUIRE ANAPHYLACTIC RESPONSIVENESS, BUT THE CONGENIC NORMAL MICE DO NOT EXHIBIT AUGMENTED RESPONSES JOURNAL OF CLINICAL INVESTIGATION Ando, A., Martin, T. R., GALLI, S. J. 1993; 92 (4): 1639-1649

    Abstract

    We treated genetically mast cell-deficient WCB6F1-Sl/Sld mice and the congenic normal (WCB6F1(-)+/+) mice with the c-kit ligand recombinant rat stem cell factor164 (rrSCF164; 100 micrograms/kg per d, subcutaneously) or with vehicle for 21 d, then passively sensitized the mice with anti-dinitrophenol30-40 immunoglobulin E (IgE) antibodies, and 1 d later measured the changes in heart rate, pulmonary dynamic compliance, and pulmonary conductance, and assessed the death rates associated with intravenous challenge of these animals with specific antigen. rrSCF164 treatment induced the development of mast cells in Sl/Sld mice, and these mice exhibited tachycardia, but not death, after challenge with IgE and antigen. rrSCF164 treatment induced mast cell hyperplasia in +/+ mice, but the cardiopulmonary changes associated with passive anaphylaxis in these mice were virtually indistinguishable from those observed in control +/+ mice treated with vehicle instead of rrSCF164. Moreover, the highest dose of antigen challenge produced significantly fewer fatalities in rrSCF164-treated than in vehicle-treated +/+ mice (1/11 vs. 8/11, respectively, P < 0.01). Thus, in normal mice, chronic treatment with rrSCF164 induces mast cell hyperplasia but does not increase, and in certain respects diminishes, the severity of IgE-dependent anaphylactic reactions.

    View details for Web of Science ID A1993MB16600009

    View details for PubMedID 7691882

  • MAST-CELLS CONTRIBUTE TO THE CHANGES IN HEART-RATE, BUT NOT HYPOTENSION OR DEATH, ASSOCIATED WITH ACTIVE ANAPHYLAXIS IN MICE JOURNAL OF IMMUNOLOGY Martin, T. R., Ando, A., Takeishi, T., Katona, I. M., Drazen, J. M., GALLI, S. J. 1993; 151 (1): 367-376

    Abstract

    The mast cell is widely thought to contribute importantly to the cardiopulmonary changes associated with anaphylaxis, but much of the evidence for this is indirect. We, therefore, performed a detailed assessment of heart rate and pulmonary function during active anaphylaxis in genetically mast cell-deficient W/Wv or S1/S1d mice, the congenic normal (+/+) mice, and W/Wv mice repaired of their mast cell deficiency by transplantation of bone marrow from the congenic +/+ mice (+/+ BM-->W/Wv mice). For all five groups of mice, Ag challenge resulted in the death of more than two-thirds of the sensitized animals, whereas none of the nonsensitized control mice died as a result of Ag infusion. Sensitized normal (WBB6F1(-)+/+ or WCB6F1(-)+/+) mice and +/+BM-->W/Wv mice developed increases in heart rate that were significantly greater than those of nonsensitized +/+ mice or those of sensitized mast cell-deficient mice, indicating that mast cells contribute to the tachycardia observed in this form of active anaphylaxis. By contrast, even though some of the pulmonary changes associated with active anaphylaxis were more severe in +/+ than in mast cell-deficient mice, it was not clear to what extent this difference was mast cell dependent. W/Wv mice undergoing active anaphylaxis developed decreases in systemic arterial blood pressure that occurred more rapidly and were more severe than those observed in the congenic +/+ mice, indicating that the hypotension associated with this model of anaphylaxis also can occur by mast cell-independent mechanisms. We conclude that in this model of anaphylaxis mast cells: 1) are required for the development of the tachycardia response; 2) may contribute to, but are not essential for, production of decreases in lung function; and 3) are not necessary for the development of hypotension or death.

    View details for Web of Science ID A1993LM57900038

    View details for PubMedID 7686942

  • HUMAN EOSINOPHILS CAN EXPRESS THE CYTOKINES TUMOR-NECROSIS-FACTOR-ALPHA AND MACROPHAGE INFLAMMATORY PROTEIN-1-ALPHA JOURNAL OF CLINICAL INVESTIGATION Costa, J. J., Matossian, K., Resnick, M. B., Beil, W. J., Wong, D. T., Gordon, J. R., Dvorak, A. M., Weller, P. F., GALLI, S. J. 1993; 91 (6): 2673-2684

    Abstract

    By in situ hybridization, 44-100% of the blood eosinophils from five patients with hypereosinophilia and four normal subjects exhibited intense hybridization signals for TNF-alpha mRNA. TNF-alpha protein was detectable by immunohistochemistry in blood eosinophils of hypereosinophilic subjects, and purified blood eosinophils from three atopic donors exhibited cycloheximide-inhibitable spontaneous release of TNF-alpha in vitro. Many blood eosinophils (39-91%) from hypereosinophilic donors exhibited intense labeling for macrophage inflammatory protein-1 alpha (MIP-1 alpha) mRNA, whereas eosinophils of normal donors demonstrated only weak or undetectable hybridization signals for MIP-1 alpha mRNA. Most tissue eosinophils infiltrating nasal polyps were strongly positive for both TNF-alpha and MIP-1 alpha mRNA. By Northern blot analysis, highly enriched blood eosinophils from a patient with the idiopathic hypereosinophilic syndrome exhibited differential expression of TNF-alpha and MIP-1 alpha mRNA. These findings indicate that human eosinophils represent a potential source of TNF-alpha and MIP-1 alpha, that levels of expression of mRNA for both cytokines are high in the blood eosinophils of hypereosinophilic donors and in eosinophils infiltrating nasal polyps, that the eosinophils of normal subjects express higher levels of TNF-alpha than MIP-1 alpha mRNA, and that eosinophils purified from the blood of atopic donors can release TNF-alpha in vitro.

    View details for Web of Science ID A1993LH12100043

    View details for PubMedID 8514874

  • DISTINCT PATTERNS OF EARLY RESPONSE GENE-EXPRESSION AND PROLIFERATION IN MOUSE MAST-CELLS STIMULATED BY STEM-CELL FACTOR, INTERLEUKIN-3, OR IGE AND ANTIGEN EUROPEAN JOURNAL OF IMMUNOLOGY Tsai, M., Tam, S. Y., GALLI, S. J. 1993; 23 (4): 867-872

    Abstract

    Stem cell factor (SCF) is encoded at the Sl locus of the mouse and is the ligand for the c-kit receptor. Recombinant rat SCF164 (rrSCF164) induces proliferation and promotes maturation of mouse mast cells in vitro and in vivo and can also induce c-kit receptor-dependent mouse mast cell degranulation. We now report that in both quiescent and non-quiescent mouse bone marrow-derived cultured mast cells (BMCMC) rrSCF164 induces increased mRNA levels for the "early response genes" c-fos, c-jun and junB but has only slight effects on the expression of junD. Recombinant mouse interleukin-3 (IL-3) also promotes proliferation of both quiescent and non-quiescent BMCMC. However, IL-3 induces increased expression of c-fos and junB only in quiescent BMCMC. Cross-linking of Fc epsilon receptor type I (Fc epsilon RI) on BMCMC by IgE and specific antigen induces a pattern of early gene expression very similar to that induced by rrSCF164. However, BMCMC stimulated through the Fc epsilon RI did not proliferate and, in comparison to control BMCMC, exhibited significantly decreased proliferation in response to rrSCF164 or IL-3. These results indicate that stimulation of BMCMC proliferation by IL-3 or rrSCF164 induces distinct patterns of early response gene expression and suggest that the proliferative effects of these growth factors may be mediated through distinct signal transduction pathways. Our data also point to previously unappreciated similarities between the effects of signaling through the c-kit receptor or the Fc epsilon RI on mast cell expression of fos and jun genes.

    View details for Web of Science ID A1993KX06500014

    View details for PubMedID 7681400

  • CLONING AND FUNCTIONAL-ANALYSIS OF THE MOUSE C-KIT PROMOTER BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS Yasuda, H., GALLI, S. J., Geissler, E. N. 1993; 191 (3): 893-901

    Abstract

    The c-kit protooncogene encodes a tyrosine kinase receptor expressed during ontogeny and adult life by several important and developmentally distinct cell lineages. Mice carrying germ line c-kit mutations exhibit deficiencies in most of these lineages, demonstrating that c-kit function is necessary for their normal development. To facilitate the identification of cis-acting elements which regulate tissue-specific c-kit expression, we cloned and characterized a mouse c-kit promoter which is functional in different cell types. A major c-kit transcription initiation site (TIS), located 58 bp upstream from the translation initiation codon, is utilized in mouse mast cells and in c-kit-positive cells in the mouse cerebellum. The effects of deletions in the 5' flanking region on reporter gene activity identify three short regulatory regions which function in both mouse and human c-kit positive cell lines. The nucleotide sequence of this region does not include CCAAT or TATA boxes but contains consensus binding sites for Sp1, Ap-2 and several short GA-rich elements which resemble binding sites for the ETS-domain proteins.

    View details for Web of Science ID A1993KV40500019

    View details for PubMedID 7682073

  • MAST-CELL ACTIVATION ENHANCES AIRWAY RESPONSIVENESS TO METHACHOLINE IN THE MOUSE JOURNAL OF CLINICAL INVESTIGATION Martin, T. R., Takeishi, T., Katz, H. R., Austen, K. F., Drazen, J. M., GALLI, S. J. 1993; 91 (3): 1176-1182

    Abstract

    Mast cell-deficient mutant mice and their normal littermates were used to determine whether activation of mast cells by anti-IgE enhances airway responsiveness to bronchoactive agonists in vivo. Pulmonary conductance was used as an index of airway response as the mice were challenged with increasing intravenous doses of methacholine (Mch) or 5-hydroxytryptamine (5-HT). Mast cell activation with anti-IgE enhanced pulmonary responsiveness to Mch in both types of normal mice (P < 0.0001 by analysis of variance) but not in either genotype of mast cell-deficient mouse. Additionally, anti-IgE pretreatment of genetically mast cell-deficient W/Wv mice whose mast cell deficiency had been repaired by infusion of freshly obtained bone marrow cells or bone marrow-derived cultured mast cells from congenic normal mice led to significant (P < 0.0001) enhancement of Mch responsiveness. 5-HT responsiveness was not significantly influenced by anti-IgE pretreatment in any of the mice studied. The data support the hypothesis that IgE-mediated activation of mast cells enhances pulmonary responsiveness to cholinergic stimulation.

    View details for Web of Science ID A1993KR46100053

    View details for PubMedID 8450046

  • New concepts about the mast cell. New England journal of medicine GALLI, S. J. 1993; 328 (4): 257-265

    View details for PubMedID 8418407

  • REVERSIBLE EXPANSION OF PRIMATE MAST-CELL POPULATIONS INVIVO BY STEM-CELL FACTOR JOURNAL OF CLINICAL INVESTIGATION GALLI, S. J., Iemura, A., Garlick, D. S., GAMBAVITALO, C., Zsebo, K. M., Andrews, R. G. 1993; 91 (1): 148-152

    Abstract

    Mast cell development in mice is critically regulated by stem cell factor (SCF), the term used here to designate a product of fibroblasts and other cell types that is a ligand for the tyrosine kinase receptor protein encoded by the proto-oncogene c-kit. However, the factors which regulate the size of mast cell populations in primates are poorly understood. Here we report that the subcutaneous administration of recombinant human SCF (rhSCF) to baboons (Papio cynocephalus) or cynomolgus monkeys (Macaca fascicularis) produced a striking expansion of mast cell populations in many anatomical sites, with numbers of mast cells in some organs of rhSCF-treated monkeys exceeding the corresponding values in control monkeys by more than 100-fold. Animals treated with rhSCF did not exhibit clinical evidence of mast cell activation, and discontinuation of treatment with rhSCF resulted in a rapid decline of mast cell numbers nearly to baseline levels. These findings are the first to demonstrate that a specific cytokine can regulate mast cell development in primates in vivo. They also provide the first evidence, in any mammalian species, to indicate that the cytokine-dependent expansion of tissue mast cell populations can be reversed when administration of the cytokine is discontinued.

    View details for Web of Science ID A1993KK04200024

    View details for PubMedID 7678600

  • THE HUMAN RECOMBINANT C-KIT RECEPTOR LIGAND, RHSCF, INDUCES MEDIATOR RELEASE FROM HUMAN CUTANEOUS MAST-CELLS AND ENHANCES IGE-DEPENDENT MEDIATOR RELEASE FROM BOTH SKIN MAST-CELLS AND PERIPHERAL-BLOOD BASOPHILS JOURNAL OF IMMUNOLOGY Columbo, M., Horowitz, E. M., Botana, L. M., MacGlashan, D. W., Bochner, B. S., Gillis, S., Zsebo, K. M., GALLI, S. J., LICHTENSTEIN, L. M. 1992; 149 (2): 599-608

    Abstract

    The gene product of the steel locus of the mouse represents a growth factor for murine mast cells and a ligand for the c-kit proto-oncogene receptor, a member of the tyrosine kinase receptor class of oncogenes (for review, see O. N. Witte. 1990. Cell 63:5). We have studied the effect of the human recombinant c-kit receptor ligand stem cell factor (rhSCF) on the release of inflammatory mediators from human skin mast cells and peripheral blood basophils and compared its activity to that of rhIL-3, rhSCF (1 ng/ml to 1 microgram/ml) activated the release of histamine and PGD2 from mast cells isolated from human skin. Analysis by digital video microscopy indicated that purified human skin mast cells (84 +/- 5% pure) responded to rhSCF (0.1 to 1 microgram/ml) challenge with a rapid, sustained rise in intracellular Ca2+ levels that was accompanied by secretion of histamine. A brief preincubation (10 min) of mast cells with rhSCF (0.1 pg/ml to 1 ng/ml) significantly enhanced (100 +/- 35%) the release of histamine induced by anti-IgE (3 micrograms/ml), but was much less effective on IgE-mediated release of PGD2. In contrast, a short term incubation with rhSCF did not potentiate the secretion of histamine activated by substance P (5 microM). A 24-h incubation of mast cells with rhSCF did not affect the release of mediators induced by anti-IgE (3 micrograms/ml), probably due to receptor desensitization, rhSCF (1 ng/ml to 3 micrograms/ml) neither caused release of histamine or leukotriene C4 (LTC4) release from leukocytes of 14 donors, nor induced a rise in intracellular Ca2+ levels in purified (greater than 70%) basophils. Brief preincubation (10 min) of leukocytes with rhSCF (1 ng/ml to 3 micrograms/ml) caused an enhancement (69 +/- 11%) of anti-IgE-induced release of histamine that was significant at concentrations as low as 3 ng/ml (p less than 0.05), whereas it appeared less effective in potentiating IgE-mediated LTC4 release. In contrast, a prolonged incubation (24 h) with rhSCF (0.1 pg/ml to 100 ng/ml) did not enhance the release of histamine or LTC4 induced by anti-IgE (0.1 microgram/ml), whereas rhIL-3 (3 ng/ml) significantly potentiated the release of both mediators.(ABSTRACT TRUNCATED AT 400 WORDS)

    View details for Web of Science ID A1992JD12200033

    View details for PubMedID 1378071

  • THE RAT C-KIT LIGAND, STEM-CELL FACTOR, INDUCES C-KIT RECEPTOR-DEPENDENT MOUSE MAST-CELL ACTIVATION INVIVO - EVIDENCE THAT SIGNALING THROUGH THE C-KIT RECEPTOR CAN INDUCE EXPRESSION OF CELLULAR FUNCTION JOURNAL OF EXPERIMENTAL MEDICINE Wershil, B. K., Tsai, M., Geissler, E. N., Zsebo, K. M., GALLI, S. J. 1992; 175 (1): 245-255

    Abstract

    Interactions between products of the mouse W locus, which encodes the c-kit tyrosine kinase receptor, and the Sl locus, which encodes a ligand for c-kit receptor, which we have designated stem cell factor (SCF), have a critical role in the development of mast cells. Mice homozygous for mutations at either locus exhibit several phenotypic abnormalities including a virtual absence of mast cells. Moreover, the c-kit ligand SCF can induce the proliferation and maturation of normal mast cells in vitro or in vivo, and also can result in repair of the mast cell deficiency of Sl/Sld mice in vivo. We now report that administration of SCF intradermally in vivo results in dermal mast cell activation and a mast cell-dependent acute inflammatory response. This effect is c-kit receptor dependent, in that it is not observed when SCF is administered to mice containing dermal mast cells expressing functionally inactive c-kit receptors, is observed with both glycosylated and nonglycosylated forms of SCF, and occurs at doses of SCF at least 10-fold lower on a molar basis than the minimally effective dose of the classical dermal mast cell-activating agent substance P. These findings represent the first demonstration in vivo that a c-kit ligand can result in the functional activation of any cellular lineage expressing the c-kit receptor, and suggest that interactions between the c-kit receptor and its ligand may influence mast cell biology through complex effects on proliferation, maturation, and function.

    View details for Web of Science ID A1992GY43600030

    View details for PubMedID 1370530

  • CYTOKINE PRODUCTION BY MAST-CELLS AND BASOPHILS CURRENT OPINION IN IMMUNOLOGY GALLI, S. J., Gordon, J. R., Wershil, B. K. 1991; 3 (6): 865-873

    Abstract

    Mast cells and/or basophils have been implicated in the expression of a wide variety of biological responses, including immediate hypersensitivity reactions, host responses to parasites and neoplasms, angiogenesis, tissue remodeling, and immunologically non-specific inflammatory and fibrotic conditions. Recent findings suggest that an important mechanism by which mast cells influence such biological responses is through the production of a broad panel of multifunctional cytokines. In contrast, the extent to which basophils can produce cytokines is uncertain.

    View details for Web of Science ID A1991HB63700005

    View details for PubMedID 1793528

  • DIFFERENCES IN THE EXPRESSION OF THE CARDIOPULMONARY ALTERATIONS ASSOCIATED WITH ANTI-IMMUNOGLOBULIN-E-INDUCED OR ACTIVE ANAPHYLAXIS IN MAST-CELL DEFICIENT AND NORMAL MICE - MAST-CELLS ARE NOT REQUIRED FOR THE CARDIOPULMONARY CHANGES ASSOCIATED WITH CERTAIN FATAL ANAPHYLACTIC RESPONSES JOURNAL OF CLINICAL INVESTIGATION Takeishi, T., Martin, T. R., Katona, I. M., Finkelman, F. D., GALLI, S. J. 1991; 88 (2): 598-608

    Abstract

    We compared the changes in heart rate (HR), pulmonary dynamic compliance (Cdyn), and pulmonary conductance (GL) associated with three different models of anaphylaxis in genetically mast cell-deficient WBB6F1-W/Wv and congenic normal (+/+) mice. Intravenous infusion of a monoclonal rat anti-mouse IgE produced a marked tachycardia, diminutions in Cdyn and GL, and death in +/+ but not W/Wv mice, and +/+ mice sensitized to develop high circulating levels of IgE exhibited HR, Cdyn, and GL responses to rat anti-IgE challenge which were significantly less intense than those in nonimmunized +/+ mice. By contrast, virtually identical cardiopulmonary responses were observed in either +/+ or W/Wv mice challenged to elicit pure active anaphylactic responses or simultaneous active and anti-IgE-dependent anaphylaxis. These findings show that anaphylactic responses associated with significant tachycardia, reductions in Cdyn and GL, and death can occur in the virtual absence of tissue mast cells. This is true even though, in normal mice, such responses are associated with extensive degranulation of tissue mast cells. By contrast, certain models of anaphylaxis, such as that induced in nonsensitized mice by anti-mouse IgE, can not be elicited in the absence of mast cells.

    View details for Web of Science ID A1991GA27600034

    View details for PubMedID 1864969

  • RELEASE OF BOTH PREFORMED AND NEWLY SYNTHESIZED TUMOR-NECROSIS-FACTOR-ALPHA (TNF-ALPHA)/CACHECTIN BY MOUSE MAST-CELLS STIMULATED VIA THE FC-EPSILON-RI - A MECHANISM FOR THE SUSTAINED ACTION OF MAST-CELL DERIVED TNF-ALPHA DURING IGE-DEPENDENT BIOLOGICAL RESPONSES JOURNAL OF EXPERIMENTAL MEDICINE Gordon, J. R., GALLI, S. J. 1991; 174 (1): 103-107

    Abstract

    Mast cell-associated mediators are generally classified into two groups: the preformed mediators, which are stored in the cells' cytoplasmic granules and are released upon exocytosis, and the newly synthesized mediators, which are not stored but are produced and secreted only after appropriate stimulation of the cell. We now report that tumor necrosis factor alpha (TNF-alpha)/cachectin represents a new type of mast cell-associated mediator, in that IgE-dependent mast cell activation results in the rapid release of preformed stores of the cytokine followed by the synthesis and sustained release of large quantities of newly formed TNF-alpha. We also demonstrate that challenge with specific antigen induces higher levels of TNF-alpha mRNA at skin sites sensitized with IgE in normal mice or mast cell-reconstituted genetically mast cell-deficient WBB6F1-W/W1' mice than at identically treated sites in WBB6F1-W/W1' mice that are devoid of mast cells. These findings identify mast cells as a biologically significant source of TNF-alpha/cachectin during IgE-dependent responses and define a mechanism whereby stimulation of mast cells via the FC epsilon RI can account for both the rapid and sustained release of this cytokine.

    View details for Web of Science ID A1991FU89700013

    View details for PubMedID 1829107

  • INDUCTION OF MAST-CELL PROLIFERATION, MATURATION, AND HEPARIN SYNTHESIS BY THE RAT C-KIT LIGAND, STEM-CELL FACTOR PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Tsai, M., Takeishi, T., Thompson, H., Langley, K. E., Zsebo, K. M., Metcalfe, D. D., Geissler, E. N., GALLI, S. J. 1991; 88 (14): 6382-6386

    Abstract

    We investigated the effects of a newly recognized multifunctional growth factor, the c-kit ligand stem cell factor (SCF), on mouse mast cell proliferation and phenotype. Recombinant rat SCF164 (rrSCF164) induced the development of large numbers of dermal mast cells in normal mice in vivo. Many of these mast cells had features of "connective tissue-type mast cells" (CTMC), in that they were reactive both with the heparin-binding fluorescent dye berberine sulfate and with safranin. In vitro, rrSCF164 induced the proliferation of cloned interleukin 3 (IL-3)-dependent mouse mast cells and primary populations of IL-3-dependent, bone marrow-derived cultured mast cells (BMCMC), which represent immature mast cells, and purified peritoneal mast cells, which represent a type of mature CTMC. BMCMC maintained in rrSCF164 not only proliferated but also matured. Prior to exposure to rrSCF164, the BMCMC were alcian blue positive, safranin negative, and berberine sulfate negative; had a histamine content of 0.08 +/- 0.02 pg per cell; and incorporated [35S]sulfate into chondroitin sulfates. After 4 wk in rrSCF164, the BMCMC were predominantly safranin positive and berberine sulfate positive, had a histamine content of 2.23 +/- 0.39 pg per cell, and synthesized 35S-labeled proteoglycans that included substantial amounts (41-70%) of [35S]heparin. These findings identify SCF as a single cytokine that can induce immature, IL-3-dependent mast cells to mature and to acquire multiple characteristics of CTMC. These findings also directly demonstrate that SCF can regulate the development of a cellular lineage expressing c-kit through effects on both proliferation and maturation.

    View details for Web of Science ID A1991FW76100091

    View details for PubMedID 1712491

  • THE RAT C-KIT LIGAND, STEM-CELL FACTOR, INDUCES THE DEVELOPMENT OF CONNECTIVE-TISSUE TYPE AND MUCOSAL MAST-CELLS INVIVO - ANALYSIS BY ANATOMICAL DISTRIBUTION, HISTOCHEMISTRY, AND PROTEASE PHENOTYPE JOURNAL OF EXPERIMENTAL MEDICINE Tsai, M., Shih, L. S., Newlands, G. F., Takeishi, T., Langley, K. E., Zsebo, K. M., Miller, H. R., Geissler, E. N., GALLI, S. J. 1991; 174 (1): 125-131

    Abstract

    Mast cell development is a complex process that results in the appearance of phenotypically distinct populations of mast cells in different anatomical sites. Mice homozygous for mutations at the W or S1 locus exhibit several phenotypic abnormalities, including a virtual absence of mast cells in all organs and tissues. Recent work indicates that W encodes the c-kit tyrosine kinase receptor, whereas S1 encodes a c-kit ligand that we have designated stem cell factor (SCF). Recombinant or purified natural forms of the c-kit ligand induce proliferation of certain mast cell populations in vitro, and injection of recombinant SCF permits mast cells to develop in mast cell-deficient WCB6F1-S1/S1d mice. However, the effects of SCF on mast cell proliferation, maturation, and phenotype in normal mice in vivo were not investigated. We now report that local administration of SCF in vivo promotes the development of connective tissue-type mast cells (CTMC) in the skin of mice and that systemic administration of SCF induces the development of both CTMC and mucosal mast cells (MMC) in rats. Rats treated with SCF also develop significantly increased tissue levels of specific rat mast cell proteases (RMCP) characteristic of either CTMC (RMCP I) or MMC (RMCP II). These findings demonstrate that SCF can induce the expansion of both CTMC and MMC populations in vivo and show that SCF can regulate at least one cellular lineage that expresses c-kit, the mast cell, through complex effects on proliferation and maturation.

    View details for Web of Science ID A1991FU89700016

    View details for PubMedID 1711559

  • MOUSE SPLENIC AND BONE-MARROW CELL-POPULATIONS THAT EXPRESS HIGH-AFFINITY FC-EPSILON RECEPTORS AND PRODUCE INTERLEUKIN-4 ARE HIGHLY ENRICHED IN BASOPHILS PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA SEDER, R. A., PAUL, W. E., Dvorak, A. M., Sharkis, S. J., KAGEYSOBOTKA, A., Niv, Y., Finkelman, F. D., BARBIERI, S. A., GALLI, S. J., Plaut, M. 1991; 88 (7): 2835-2839

    Abstract

    Splenic and bone marrow cells from normal mice, and from mice that have been polyclonally activated by injection of anti-IgD antibody, contain cells that produce interleukin 4 (IL-4) in response to crosslinkage of Fc epsilon receptors (Fc epsilon R) or Fc gamma R or to ionomycin. Isolated Fc epsilon R+ cells have recently been shown to contain all of the IL-4-producing capacity of the nonlymphoid compartment of spleen and bone marrow. Here, purified Fc epsilon R+ cells are shown to be enriched in cells that contain histamine and express alcian blue-positive cytoplasmic granules. By electron microscopy, the vast majority of cytoplasmic granule-containing cells are basophils; they constitute approximately 25% and approximately 50%, respectively, of Fc epsilon R+ spleen and bone marrow cells from anti-IgD-injected mice. The Fc epsilon R- populations contain cells that form colonies typical of mast cells. The Fc epsilon R+ populations also contain cells that, upon culture with IL-3, form colonies of alcian blue-positive cells, but (in contrast to colonies derived from Fc epsilon R- populations) the colonies are small, and all the cells die within 2-3 weeks. The Fc epsilon R+ cells synthesize histamine during a 60-hr culture with IL-3, while the Fc epsilon R- cells do not. These results indicate that IL-4-producing Fc epsilon R+ cells are highly enriched in basophils.

    View details for Web of Science ID A1991FE86400047

    View details for PubMedID 1826367

  • STEM-CELL FACTOR (SCF), A NOVEL HEMATOPOIETIC GROWTH-FACTOR AND LIGAND FOR C-KIT TYROSINE KINASE RECEPTOR, MAPS ON HUMAN-CHROMOSOME 12 BETWEEN 12Q14.3 AND 12QTER SOMATIC CELL AND MOLECULAR GENETICS Geissler, E. N., Liao, M., BROOK, J. D., Martin, F. H., Zsebo, K. M., Housman, D. E., GALLI, S. J. 1991; 17 (2): 207-214

    Abstract

    Recently a novel hematopoietic growth factor, stem cell factor (SCF), was cloned and demonstrated to be the ligand for the c-kit tyrosine kinase receptor. In the mouse, SCF is encoded by Sl (steel), a gene critical to the development of several distinct cell lineages during embryonic life and which has important effects on hematopoiesis in the adult animal. The Sl/SCF locus maps to the distal region of mouse chromosome 10, in the vicinity of genes that have been mapped to human chromosome 12. Here we report the use of somatic cell hybrid lines to localize SCF to the long arm of human chromosome 12, between 12q14.3 and 12qter. In addition to localizing the Sl homolog in man, these data provide further evidence for the conservation of synteny between the long arm of human chromosome 12 and the distal end of mouse chromosome 10.

    View details for Web of Science ID A1991FD59400010

    View details for PubMedID 1707188

  • RECRUITMENT OF NEUTROPHILS DURING IGE-DEPENDENT CUTANEOUS LATE PHASE REACTIONS IN THE MOUSE IS MAST CELL-DEPENDENT - PARTIAL INHIBITION OF THE REACTION WITH ANTISERUM AGAINST TUMOR-NECROSIS-FACTOR-ALPHA JOURNAL OF CLINICAL INVESTIGATION Wershil, B. K., Wang, Z. S., Gordon, J. R., GALLI, S. J. 1991; 87 (2): 446-453

    Abstract

    Much of the clinically important pathology associated with IgE-dependent disorders is thought to reflect the actions of the blood-borne leukocytes recruited during these responses. To evaluate the extent to which mast cells are responsible for the leukocyte infiltration associated with IgE-dependent cutaneous reactions, we attempted to elicit these responses in normal mice, genetically mast cell-deficient W/Wv mice, and in W/Wv mice selectively repaired of their mast cell deficiency by the intradermal injection of cultured mast cells derived from the congenic normal (+/+) mice. We found that the tissue swelling associated with IgE-dependent passive cutaneous anaphylaxis reactions developed rapidly and diminished markedly from 2 to 4 h after antigen challenge, but remained detectable for at least 24 h after elicitation of the responses. Infiltration of leukocytes (predominantly neutrophils) also occurred at these sites, but reached maximal levels 6-12 h after antigen challenge, persisted at high levels for 24 h, and largely waned by 48 h. Virtually all of the tissue swelling and leukocyte infiltration associated with IgE-dependent cutaneous reactions was mast cell dependent. Intradermal injection of 40 U of recombinant murine TNF-alpha (rmTNF-alpha) elicited neutrophil infiltration similar in magnitude and kinetics to that observed after IgE-dependent mast cell degranulation. A rabbit anti-rmTNF-alpha (R anti-rmTNF-alpha) antiserum, which was able to inhibit 84% of the neutrophil infiltration observed after i.d. injection of rmTNF-alpha, inhibited IgE-, and mast cell-dependent leukocyte infiltration by 47 +/- 7% in three separate experiments. These findings indicate that TNF-alpha contributes to mast cell-dependent recruitment of leukocytes during IgE-dependent cutaneous late phase reactions, but suggest that other mast cell-associated mediators probably also contribute to this response.

    View details for Web of Science ID A1991EW29500010

    View details for PubMedID 1991831

  • ROLE OF MAST-CELLS IN ION-TRANSPORT ABNORMALITIES ASSOCIATED WITH INTESTINAL ANAPHYLAXIS - CORRECTION OF THE DIMINISHED SECRETORY RESPONSE IN GENETICALLY MAST CELL-DEFICIENT W/WV MICE BY BONE-MARROW TRANSPLANTATION JOURNAL OF CLINICAL INVESTIGATION Perdue, M. H., Masson, S., Wershil, B. K., GALLI, S. J. 1991; 87 (2): 687-693

    Abstract

    To investigate the role of mast cells in transport abnormalities during intestinal anaphylaxis, we examined responses to antigen in isolated intestinal preparations from ovalbumin-sensitized genetically mast cell-deficient WBB6F1-W/Wv (W/Wv) mice and congenic normal WBBGF1(-)+/+ (+/+) mice. Changes in ion transport (primarily secretion of chloride ions) were indicated by increases in short-circuit current (Isc). In tissues from +/+ mice, antigen caused increases in Isc which were significantly inhibited by antagonists to histamine (diphenhydramine) and serotonin (ketanserin), by a cyclooxygenase inhibitor (piroxicam) and by a neurotoxin (tetrodotoxin). In preparations from W/Wv mice, antigen-stimulated responses were approximately 30% of that in +/+ mice and were inhibited only by piroxicam. Responses to electrical transmural stimulation of nerves were approximately 50% in W/Wv versus +/+ mice, and were inhibited by antagonists of mast cell mediators in +/+ but not W/Wv mice. Reconstitution of mast cells in W/Wv mice by intravenous injection of +/+ bone marrow cells restored the normal responses to both antigen and nerve stimulation. Our results indicate that mast cell-dependent mechanisms are primarily responsible for the ion secretion associated with intestinal anaphylaxis, but that other cells are also involved. In addition, our data provide evidence for the functional importance of bidirectional communication between nerves and mast cells in the regulation of ion transport in the gastrointestinal tract.

    View details for Web of Science ID A1991EW29500041

    View details for PubMedID 1991852

  • [I-125] FIBRIN DEPOSITION OCCURS AT BOTH EARLY AND LATE INTERVALS OF IGE-DEPENDENT OR CONTACT SENSITIVITY REACTIONS ELICITED IN MOUSE SKIN - MAST CELL-DEPENDENT AUGMENTATION OF FIBRIN DEPOSITION AT EARLY INTERVALS IN COMBINED IGE-DEPENDENT AND CONTACT SENSITIVITY REACTIONS JOURNAL OF IMMUNOLOGY Mekori, Y. A., GALLI, S. J. 1990; 145 (11): 3719-3727

    Abstract

    When elicited in the skin of mice, either IgE-dependent immediate hypersensitivity reactions or T cell-dependent contact sensitivity (CS) reactions result in local extravasation of [125I]fibrinogen and deposition of [125I]fibrin. However, these two types of reaction differ in kinetics and in requirement for IgE, mast cells, or T cells. In the present study, we investigated the kinetics and magnitude of [125I]fibrin deposition in combined IgE-dependent and CS reactions elicited simultaneously at the same site and compared the results with those obtained when the two reactions were elicited at separate sites. We found that [125I]fibrin deposition in pure IgE-dependent reactions was greater at 2 or 6 h after challenge than at 24 h, but that significant fibrin deposition persisted at those sites 24 h after challenge. In CS reactions, [125I]fibrin deposition was detected as early as 2 h after challenge, indicating that fibrin deposition accompanies the "early component" of CS detected by Van Loveren et al. with the use of measurements of tissue swelling. But much more [125I]fibrin deposition was present in CS reactions at 24 h than at 2 or 6 h after Ag challenge. When IgE-dependent and CS reactions were elicited at the same site, [125I]fibrin deposition at early intervals (2 to 6 h) after challenge was increased three- to 25-fold compared with that seen in isolated CS reactions, but at 24 h the results in the combined reactions were virtually identical to those in CS responses. Studies in genetically mast cell-deficient and congenic normal mice indicated that mast cells were required for expression of the IgE-dependent augmentation of [125I]fibrin deposition observed at early intervals in combined IgE-dependent and CS reactions, but not for the [125I]fibrin deposition associated with "pure" CS reactions. These findings indicate that the net effect of IgE-dependent mast cell activation on CS responses is to increase the fibrin deposition associated with these responses, but this effect is appreciated only at early intervals after elicitation of the reaction.

    View details for Web of Science ID A1990EK39100026

    View details for PubMedID 2246510

  • MAST-CELLS AS A SOURCE OF MULTIFUNCTIONAL CYTOKINES IMMUNOLOGY TODAY Gordon, J. R., Burd, P. R., GALLI, S. J. 1990; 11 (12): 458-464

    Abstract

    Mast cells have been implicated in the expression of a wide variety of biological responses, including immediate hypersensitivity reactions, host responses to parasites and neoplasms, immunologically non-specific inflammatory and fibrotic conditions, angiogenesis, and tissue remodeling and wound healing. However, the molecular basis for the action of the mast cell in many of these responses is obscure. In this review, John Gordon, Parris Burd and Stephen Galli suggest that the production of a broad panel of multifunctional cytokines may represent an important mechanism by which mast cells influence physiological, immunological and pathological processes.

    View details for Web of Science ID A1990EN94000013

    View details for PubMedID 2073318

  • HUMAN EOSINOPHILS EXPRESS TRANSFORMING GROWTH FACTOR-ALPHA JOURNAL OF EXPERIMENTAL MEDICINE Wong, D. T., Weller, P. F., GALLI, S. J., Elovic, A., RAND, T. H., Gallagher, G. T., Chiang, T., Chou, M. Y., Matossian, K., McBride, J., Todd, R. 1990; 172 (3): 673-681

    Abstract

    Transforming growth factor alpha (TGF-alpha) is a pleuripotential cytokine with diverse biological effects, including the ability to influence the proliferation of normal cells or neoplastic epithelial cells. Eosinophils are a subset of granulocytes that normally enter the peripheral tissues, particularly those beneath gastrointestinal, respiratory, and urogenital epithelium, where they reside in close proximity to the epithelial elements. In this study, we demonstrate that the great majority of eosinophils infiltrating the interstitial tissues adjacent to two colonic adenocarcinomas and two oral squamous cell carcinomas labeled specifically by in situ hybridization with a 35S-riboprobe for human TGF-alpha (hTGF-alpha). No other identifiable leukocytes in these lesions contained detectable hTGF-alpha mRNA. We also examined leukocytes purified from a patient with the idiopathic hypereosinophilic syndrome. 80% of these eosinophils, but none of the patient's neutrophils or mononuclear cells, were positive for hTGF-alpha mRNA by in situ hybridization, and 55% of these eosinophils were positive by immunohistochemistry with a monoclonal antibody directed against the COOH terminus of the mature hTGF-alpha peptide. Finally, the identification of the purified eosinophil-associated transcript as hTGF-alpha was confirmed by polymerase chain reaction product restriction enzyme analysis followed by Southern blot hybridization. In contrast to eosinophils from the patient with hypereosinophilic syndrome, the peripheral blood eosinophils from only two of seven normal donors had detectable TGF-alpha mRNA and none of these eosinophils contained immunohistochemically detectable TGF-alpha product. Taken together, these findings establish that human eosinophils can express TGF-alpha, but suggest that the expression of TGF-alpha by eosinophils may be under microenvironmental regulation. Demonstration of TGF-alpha production by tissue-infiltrating eosinophils and the eosinophils in the hypereosinophilic syndrome identifies a novel mechanism by which eosinophils might contribute to physiological, immunological, and pathological responses.

    View details for Web of Science ID A1990DW49100001

    View details for PubMedID 1696954

  • MAST-CELLS AS A SOURCE OF BOTH PREFORMED AND IMMUNOLOGICALLY INDUCIBLE TNF-ALPHA CACHECTIN NATURE Gordon, J. R., GALLI, S. J. 1990; 346 (6281): 274-276

    Abstract

    Tumour necrosis factor-alpha (TNF-alpha)/cachectin is a multifunctional cytokine that has effects in inflammation, sepsis, lipid and protein metabolism, haematopoiesis, angiogenesis and host resistance to parasites and malignancy. TNF-alpha was first described in activated macrophages, but certain mouse or rat mast cell populations (reviewed in refs 4,5) and some in vitro-derived human cells with cytochemical features of mast cells-basophils may also contain products similar to TNF-alpha. Here we present evidence that resident mouse peritoneal mast cells constitutively contain large amounts of TNF-alpha bioactivity, whereas cultured, immature mast cells vary in their TNF-alpha content. IgE-dependent activation of cultured or peritoneal mast cells induces extracellular release of TNF-alpha and augments levels of TNF-alpha messenger RNA and bioactivity. These findings identify mouse mast cells as an important source of both preformed and immunologically inducible TNF-alpha, and suggest that release of TNF-alpha by mast cells may contribute to host defence, the pathophysiology of allergic diseases and other processes dependent on TNF-alpha.

    View details for Web of Science ID A1990DP45600067

    View details for PubMedID 2374592

  • ANATOMICAL VARIATION IN MAST-CELL NERVE ASSOCIATIONS IN THE RAT SMALL-INTESTINE, HEART, LUNG, AND SKIN - SIMILARITIES OF DISTANCES BETWEEN NEURAL PROCESSES AND MAST-CELLS, EOSINOPHILS, OR PLASMA-CELLS IN THE JEJUNAL LAMINA PROPRIA LABORATORY INVESTIGATION Arizono, N., Matsuda, S., Hattori, T., Kojima, Y., Maeda, T., GALLI, S. J. 1990; 62 (5): 626-634

    Abstract

    Several studies have indicated that mast cells occur in close proximity to enteric nerves in the gastrointestinal tract of rats, man, and other mammalian species, and such intimate associations have been proposed as one of the anatomical bases of communication between the immune and the nervous systems. However, the specificity of anatomical associations between enteric nerves and mast cells, as opposed to other bone marrow-derived or lymphoid cells normally present in mucosal sites, is unclear. We used transmission electron microscopy to quantify the distances between mast cells and neural processes (nerve terminals or axons) in the small intestinal mucosa, right atrium, skin, and pulmonary parenchyma of normal rats, and in the small intestinal mucosa and lung parenchyma of rats that had undergone hyperplasia of the mast cell populations in these sites as a result of infection with the nematode Nippostrongylus brasiliensis. In the jejunal mucosa of normal rats, 8.0% of mast cells occurred within 100 nm of neural processes and an additional 11.0% between 101 and 500 nm of these structures; the corresponding figures for eosinophils were 3.3% (N.S. vs. mast cell value) and 23.3% (p less than 0.05 vs. mast cell value) and for plasma cells were 8.5% and 14.6% (N.S. vs. mast cell values). In the right atrium, 1.2% of mast cells occurred within 100 nm and an additional 13.4% within 101 and 500 nm of neural processes, whereas no mast cells were observed within 500 nm of neural processes in the pulmonary parenchyma or ear skin. Infection with N. brasiliensis increased by 61% the proportion of mast cells within 500 nm of neural processes in the jejunal mucosa and resulted in the appearance of mast cells in close association with these structures in the jejunal muscularis propria, but had no effect on the proportion of mast cells within 100 or 500 nm of neural processes in the pulmonary parenchyma. Acetylcholine esterase staining demonstrated dense networks of neural processes in the three sites where some mast cells were closely associated with these structures (jejunal mucosa and muscularis, right atrium) but not in the pulmonary parenchyma or ear skin. Taken together, our findings indicate that mast cells occur in close proximity to neural processes in sites where these structures are abundant, but that anatomical associations as close as those between mast cells and neural processes can also occur between such structures and other bone marrow-derived cells (eosinophils) or lymphoid cells (plasma cells) resident in the small intestinal mucosa.(ABSTRACT TRUNCATED AT 400 WORDS)

    View details for Web of Science ID A1990DF15000012

    View details for PubMedID 2342332

  • PHORBOL 12-MYRISTATE 13-ACETATE INDUCED DEVELOPMENT OF FUNCTIONALLY ACTIVE MAST-CELLS IN W/WV BUT NOT SL/SLD GENETICALLY MAST CELL-DEFICIENT MICE BLOOD Gordon, J. R., GALLI, S. J. 1990; 75 (8): 1637-1645

    Abstract

    The normal skin and other tissues of adult genetically mast cell-deficient WBB6F1-W/Wv or WCB6F1-Sl/Sld mice contain less than 1.0% the number of mast cells present in the corresponding tissues of the congenic normal (+/+) mice. We previously reported that mature dermal mast cells developed locally in the skin of W/Wv, but not Sl/Sld, mice at sites of chronic idiopathic dermatitis. We now report that the repeated application of phorbol 12-myristate 13-acetate (PMA) to the ear skin of either W/Wv or +/+ mice induces both dermatitis and a striking and dose-dependent increase in the number of dermal mast cells. The number of dermal mast cells at sites treated for 6 weeks with 5 micrograms PMA, three times per week, was 39 +/- 7/mm2 and 305 +/- 34/mm2 for W/Wv and +/+ mice, respectively; the corresponding values for vehicle-treated skin were 1.5 +/- 1.0/mm2 and 145 +/- 8/mm2, respectively. The PMA-induced dermal mast cells in W/Wv mice appeared mature by morphology, stained with the heparin-binding fluorescent dye, berberine sulfate, and were competent to express IgE-dependent passive cutaneous anaphylaxis responses. The development of mast cells was a local, not systemic, effect of PMA treatment. PMA treatment also induced dermatitis in both WCB6F1-Sl/Sld and +/+ mice, but was associated with increased numbers of dermal mast cells only in the WCB6F1(-)+/+ mice. PMA treatment had no detectable effect on the ability of bone marrow-derived cultured mast cells to survive in the skin of Sl/Sld mice. These findings establish a convenient model system for analyzing factors associated with the development of endogenous populations of mast cells in genetically mast cell-deficient W/Wv mice.

    View details for Web of Science ID A1990CZ67500008

    View details for PubMedID 2328315

  • New insights into "the riddle of the mast cells": microenvironmental regulation of mast cell development and phenotypic heterogeneity. Laboratory investigation; a journal of technical methods and pathology GALLI, S. J. 1990; 62 (1): 5-33

    View details for PubMedID 2404155

  • SUBSTANCE-P-INDUCED AUGMENTATION OF CUTANEOUS VASCULAR-PERMEABILITY AND GRANULOCYTE INFILTRATION IN MICE IS MAST-CELL DEPENDENT JOURNAL OF CLINICAL INVESTIGATION Yano, H., Wershil, B. K., Arizono, N., GALLI, S. J. 1989; 84 (4): 1276-1286

    Abstract

    The undecapeptide substance P is thought to mediate both vasodilatation and augmented vascular permeability when released from sensory nerve endings in the skin. Substance P also induces mast cell degranulation in vitro or in vivo. However, the extent to which substance P-induced changes in vascular permeability are mast cell-dependent is unclear. We investigated this issue by injecting substance P and certain related peptides (substance P1-4, substance P4-11) into the skin of genetically mast cell-deficient WBB6F1-W/W or WCB6F1- SI/SId mice the congenic normal (+/+) mice, and W/W mice which had undergone selective local repair of their mast cell deficiency by intradermal injection of IL-3-dependent mast cells generated in vitro from the bone marrow cells of the congenic +/+ mice. Substance P induced significant augmentation of vascular permeability and significant cutaneous swelling when injected into normal mice at doses as low as 2 pmol i.d. Substance P also induced granulocyte infiltration, although the infiltrate were modest and were seen at doses of peptide from 5 to more than 20-fold higher than those required for induction of tissue swelling. The effects of substance P on tissue swelling, vascular permeability, and granulocyte infiltration were virtually entirely mast cell dependent. By contrast, substance P1-4 was inactive in our assays at 25 nmol/site, and substance P4-11 induced modest augmentation of vascular permeability, which was at least in part mast cell independent.

    View details for Web of Science ID A1989AT80800031

    View details for PubMedID 2477394

  • ANALYSIS OF MAST-CELL FUNCTION IN BIOLOGICAL RESPONSES NOT INVOLVING IGE INTERNATIONAL ARCHIVES OF ALLERGY AND APPLIED IMMUNOLOGY GALLI, S. J., Wershil, B. K., Mekori, Y. A. 1987; 82 (3-4): 269-271

    Abstract

    Mast cells are critical for the expression of certain IgE-mediated responses, but the precise contributions of mast cells to biological processes not involving IgE are obscure. We have employed genetically mast cell-deficient WBB6F1-W/Wv and WCB6F1-S1/S1d mice to investigate the roles of mast cells in several different biological responses. This work strongly suggests that mast cells are not required for the elicitation of contact sensitivity (CS) responses, suppressor T cell-dependent tolerance to CS, reserpine-induced inhibition of CS responses, or bleomycin-induced pulmonary fibrosis. By contrast, mast cells appear to contribute to the acute gastric injury induced by ethanol and the acute inflammation of the skin induced by croton oil.

    View details for Web of Science ID A1987G942800009

    View details for PubMedID 3570498

Conference Proceedings


  • Second symposium on the definition and management of anaphylaxis: summary report--Second National Institute of Allergy and Infectious Disease/Food Allergy and Anaphylaxis Network symposium. Sampson, H. A., Mu˝oz-Furlong, A., Campbell, R. L., Adkinson, N. F., Bock, S. A., Branum, A., Brown, S. G., Camargo, C. A., Cydulka, R., Galli, S. J., Gidudu, J., Gruchalla, R. S., Harlor, A. D., Hepner, D. L., Lewis, L. M., Lieberman, P. L., Metcalfe, D. D., O'Connor, R., Muraro, A., Rudman, A., Schmitt, C., Scherrer, D., Simons, F. E., Thomas, S., Wood, J. P., Decker, W. W. 2006: 391-397

    Abstract

    There is no universal agreement on the definition of anaphylaxis or the criteria for diagnosis. In July 2005, the National Institute of Allergy and Infectious Disease and Food Allergy and Anaphylaxis Network convened a second meeting on anaphylaxis, which included representatives from 16 different organizations or government bodies, including representatives from North America, Europe, and Australia, to continue working toward a universally accepted definition of anaphylaxis, establish clinical criteria that would accurately identify cases of anaphylaxis with high precision, further review the evidence on the most appropriate management of anaphylaxis, and outline the research needs in this area.

    View details for PubMedID 16461139

  • Probing the roles of mast cells and basophils in natural and acquired immunity, physiology and disease. Marone, G., Galli, S. J., Kitamura, Y. 2002: 425-427

    View details for PubMedID 12200056

Footer Links:

Stanford Medicine Resources: