Professional Education

  • Bachelor of Science, Cornell University (2006)
  • Doctor of Philosophy, Cornell University (2012)

Stanford Advisors


Journal Articles

  • Barcode-Enabled Sequencing of Plasmablast Antibody Repertoires in Rheumatoid Arthritis ARTHRITIS & RHEUMATOLOGY Tan, Y., Kongpachith, S., Blum, L. K., Ju, C., Lahey, L. J., Lu, D. R., Cai, X., Wagner, C. A., Lindstrom, T. M., Sokolove, J., Robinson, W. H. 2014; 66 (10): 2706-2715

    View details for DOI 10.1002/art.38754

    View details for Web of Science ID 000342744300008

  • High-throughput sequencing of natively paired antibody chains provides evidence for original antigenic sin shaping the antibody response to influenza vaccination CLINICAL IMMUNOLOGY Tan, Y., Blum, L. K., Kongpachith, S., Ju, C., Cai, X., Lindstrom, T. M., Sokolove, J., Robinson, W. H. 2014; 151 (1): 55-65


    We developed a DNA barcoding method to enable high-throughput sequencing of the cognate heavy- and light-chain pairs of the antibodies expressed by individual B cells. We used this approach to elucidate the plasmablast antibody response to influenza vaccination. We show that >75% of the rationally selected plasmablast antibodies bind and neutralize influenza, and that antibodies from clonal families, defined by sharing both heavy-chain VJ and light-chain VJ sequence usage, do so most effectively. Vaccine-induced heavy-chain VJ regions contained on average >20 nucleotide mutations as compared to their predicted germline gene sequences, and some vaccine-induced antibodies exhibited higher binding affinities for hemagglutinins derived from prior years' seasonal influenza as compared to their affinities for the immunization strains. Our results show that influenza vaccination induces the recall of memory B cells that express antibodies that previously underwent affinity maturation against prior years' seasonal influenza, suggesting that 'original antigenic sin' shapes the antibody response to influenza vaccination.

    View details for DOI 10.1016/j.clim.2013.12.008

    View details for Web of Science ID 000332351100006

  • Intestinal infection with Trichinella spiralis induces distinct, regional immune responses. Veterinary parasitology Blum, L. K., Mohanan, S., Fabre, M. V., Yafawi, R. E., Appleton, J. A. 2013; 194 (2-4): 101-105


    The aim of this study was to evaluate differences between the small and large intestines (SI and LI) with regard to colonization and immunity during infection with Trichinella spiralis. In orally infected C57BL/6 mice, the gender ratios of worms differed among the SI, cecum, and LI. Mucosal mastocytosis developed in the SI but not in the LI, consistent with reduced IL-9 and IL-13 production by explants from the LI. Despite these differences, worms were cleared at the same rate from both sites. Furthermore, IL-10 production was reduced in the LI, yet it was instrumental in limiting local inflammation. Finally, passive immunization of rat pups with tyvelose-specific antibodies effectively cleared fist-stage larvae from all intestinal regions. We conclude that despite regional differences in immune responsiveness and colonization, immune mechanisms that clear T. spiralis operate effectively throughout the intestinal tract.

    View details for DOI 10.1016/j.vetpar.2013.01.030

    View details for PubMedID 23465441

  • Participation of MyD88 and interleukin-33 as innate drivers of Th2 immunity to Trichinella spiralis. Infection and immunity Scalfone, L. K., Nel, H. J., Gagliardo, L. F., Cameron, J. L., Al-Shokri, S., Leifer, C. A., Fallon, P. G., Appleton, J. A. 2013; 81 (4): 1354-1363


    Trichinella spiralis is a highly destructive parasitic nematode that invades and destroys intestinal epithelial cells, injures many different tissues during its migratory phase, and occupies and transforms myotubes during the final phase of its life cycle. We set out to investigate the role in immunity of innate receptors for potential pathogen- or danger-associated molecular patterns (PAMPs or DAMPs). Focusing on the MyD88-dependent receptors, which include Toll-like receptors (TLRs) and interleukin-1 (IL-1) family members, we found that MyD88-deficient mice expelled worms normally, while TLR2/4-deficient mice showed accelerated worm expulsion, suggesting that MyD88 was active in signaling pathways for more than one receptor during intestinal immunity. A direct role for PAMPs in TLR activation was not supported in a transactivation assay involving a panel of murine and human TLRs. Mice deficient in the IL-1 family receptor for the DAMP, IL-33 (called ST2), displayed reduced intestinal Th2 responses and impaired mast cell activation. IL-33 was constitutively expressed in intestinal epithelial cells, where it became concentrated in nuclei within 2 days of infection. Nuclear localization was an innate response to infection that occurred in intestinal regions where worms were actively migrating. Th2 responses were also compromised in the lymph nodes draining the skeletal muscles of ST2-deficient mice, and this correlated with increased larval burdens in muscle. Our results support a mechanism in which the immune system recognizes and responds to tissue injury in a way that promotes Th2 responses.

    View details for DOI 10.1128/IAI.01307-12

    View details for PubMedID 23403558

  • In vitro modelling of rat mucosal mast cell function in Trichinella spiralis infection. Parasite immunology Thrasher, S. M., Scalfone, L. K., Holowka, D., Appleton, J. A. 2013; 35 (1): 21-31


    Intestinal infection with the parasitic nematode, Trichinella spiralis, provides a robust context for the study of mucosal mast cell function. In rats, mucosal mast cells are exposed to parasites during the earliest stage of infection, affording an opportunity for mast cells to contribute to an innate response to infection. During secondary infection, degranulation of rat mucosal mast cells coincides with expulsion of challenge larvae from the intestine. The goal of this study was to evaluate the rat bone marrow-derived mast cells (BMMC) and the rat basophilic leukaemia cell line (RBL-2H3) as models for mucosal mast cells, using parasite glycoproteins and antibody reagents that have been tested extensively in rats in vivo. We found that BMMC displayed a more robust mucosal phenotype. Although T. spiralis glycoproteins bound to mast cell surfaces in the absence of antibodies, they did not stimulate degranulation, nor did they inhibit degranulation triggered by immune complexes. Parasite glycoproteins complexed with specific monoclonal IgGs provoked release of rat mast cell protease II (RMCPII) and ?-hexosaminidase from both cell types in a manner that replicated results observed previously in passively immunized rats. Our results document that RBL-2H3 cells and BMMC model rat mucosal mast cells in the contexts of innate and adaptive responses to T. spiralis.

    View details for DOI 10.1111/pim.12014

    View details for PubMedID 23094823

  • Expulsion of Secondary Trichinella spiralis Infection in Rats Occurs Independently of Mucosal Mast Cell Release of Mast Cell Protease II JOURNAL OF IMMUNOLOGY Blum, L. K., Thrasher, S. M., Gagliardo, L. F., Fabre, V., Appleton, J. A. 2009; 183 (9): 5816-5822


    Our aim was to elucidate the contribution of mucosal mast cells to the effector phase of a secondary immune response to Trichinella spiralis. During secondary infection, rats expel 90-99% of T. spiralis first-stage larvae from the intestine in a matter of hours. This phenomenon appears to be unique to rats and has been called rapid expulsion. Primary intestinal infection by T. spiralis induces mastocytosis, and mast cell degranulation occurs when challenged rats exhibit rapid expulsion. These observations have engendered the view that mast cells mediate rapid expulsion. In this study, we report that immunization of adult Albino Oxford rats by an infection limited to the muscle phase did not induce intestinal mastocytosis, yet such rats exhibited rapid expulsion when challenged orally. Although mastocytosis was absent, the protease unique to mucosal mast cells, rat mast cell protease II (RMCPII), was detected in sera at the time of expulsion. We further evaluated mast cell activity in neonatal rats that display rapid expulsion. Pups born to infected dams displayed rapid expulsion, and RMCPII was detected in their sera. By feeding pups parasite-specific mAbs or polyclonal Abs before challenge infection, it was possible to dissociate mast cell degranulation from parasite expulsion. These results indicate that rapid expulsion can occur in the absence of either intestinal mastocytosis or RMCPII release. Furthermore, release of RMCPII is not sufficient to cause expulsion. The data argue against a role for mast cells in the mechanism underlying the effector phase of protective immunity against T. spiralis in rats.

    View details for DOI 10.4049/jimmunol.0900944

    View details for Web of Science ID 000271488500044

    View details for PubMedID 19812197

  • TLR9 traffics through the Golgi complex to localize to endolysosomes and respond to CpG DNA IMMUNOLOGY AND CELL BIOLOGY Chockalingam, A., Brooks, J. C., Cameron, J. L., Blum, L. K., Leifer, C. A. 2009; 87 (3): 209-217


    Toll-like receptor 9 (TLR9) promiscuously binds self- and microbial DNA, but only microbial DNA elicits an inflammatory response. How TLR9 discriminates between self- and foreign DNA is unclear, but inappropriate localization of TLR9 permits response to self-DNA, suggesting that TLR9 localization and trafficking are critical components. The molecular mechanisms controlling the movement of TLR9 may provide new insight into the recognition of DNA in normal and in pathological conditions such as autoimmune systemic lupus erythematosus. We have shown earlier that TLR9 is retained in the endoplasmic reticulum (ER) and it moves to endolysosomes to recognize CpG DNA. Other studies have suggested that TLR9 bypasses the Golgi complex to access endolysosomes. Here, we show that TLR9 translocates from ER to endolysosomes through the Golgi complex and that Golgi export is required for optimal TLR9 signaling. In all, 6-13% of TLR9 constitutively exits the ER, moves through the Golgi complex and resides in lysosomal-associated membrane protein-1-positive vesicles. TLR9 bound to CpG DNA had glycan modifications indicative of Golgi processing confirming that TLR9 travels through the Golgi complex to access CpG DNA in endolysosomes. Together, these data support a model where TLR9 uses traditional secretory pathways and does not bypass the Golgi complex.

    View details for DOI 10.1038/icb.2008.101

    View details for Web of Science ID 000264748100005

    View details for PubMedID 19079358

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