High-Parametric Evaluation of Human Invariant Natural Killer T Cells to Delineate Heterogeneity in Allo- and Autoimmunity.
Human invariant natural killer T cells (iNKTs) are a rare innate-like lymphocyte population that recognize glycolipids presented on CD1d. Studies in mice have shown that these cells are heterogenous and capable of enacting diverse functions, and the composition of iNKT subsets can alter disease outcomes. In contrast, far less is known about how heterogeneity in human iNKTs relates to disease. To address this, we use a high-dimensional, data-driven approach to devise a framework to parse human iNKT heterogeneity. Our data revealed novel and previously described iNKT phenotypes with distinct functions. In particular, we found two phenotypes of interest: 1) a population with Th1 function that was increased with iNKT activation characterized by HLA-II+CD161- expression, and 2) a population with enhanced cytotoxic function characterized by CD4-CD94+ expression. These populations, respectively, correlate with acute graft-versus-host disease after allogeneic hematopoietic stem cell transplantation and with new onset type 1 diabetes. Our study identifies human iNKT phenotypes associated with human disease that could aid in the development of biomarkers or therapeutics targeting iNKTs.
View details for DOI 10.1182/blood.2019001903
View details for PubMedID 31935280
Hyaluronan synthesis inhibition impairs antigen presentation and delays transplantation rejection.
Matrix biology : journal of the International Society for Matrix Biology
A coat of pericellular hyaluronan surrounds mature dendritic cells (DC) and contributes to cell-cell interactions. We asked whether 4-methylumbelliferone (4MU), an oral inhibitor of HA synthesis, could inhibit antigen presentation. We find that 4MU treatment reduces pericellular hyaluronan, destabilizes interactions between DC and T-cells, and prevents T-cell proliferation in vitro and in vivo. These effects were observed only when 4MU was added prior to initial antigen presentation but not later, consistent with 4MU-mediated inhibition of de novo antigenic responses. Building on these findings, we find that 4MU delays rejection of allogeneic pancreatic islet transplant and allogeneic cardiac transplants in mice and suppresses allogeneic T-cell activation in human mixed lymphocyte reactions. We conclude that 4MU, an approved drug, may have benefit as an adjunctive agent to delay transplantation rejection.
View details for DOI 10.1016/j.matbio.2020.12.001
View details for PubMedID 33290836
- Donor-Derived Cytokine-Induced Killer Cell Infusion as Consolidation after Nonmyeloablative Allogeneic Transplantation for Myeloid Neoplasms BIOLOGY OF BLOOD AND MARROW TRANSPLANTATION 2019; 25 (7): 1293?1303
- Transplantation of donor grafts with defined ratio of conventional and regulatory T cells in HLA-matched recipients JCI INSIGHT 2019; 4 (10)
4-Methylumbelliferyl glucuronide contributes to hyaluronan synthesis inhibition.
The Journal of biological chemistry
4-Methylumbelliferone (4-MU) inhibits hyaluronan (HA) synthesis and is an approved drug used for managing biliary spasm. However, rapid and efficient glucuronidation is thought to limit its utility for systemically inhibiting HA synthesis. In particular, 4-MU in mice has a short half-life causing most of the drug to be present as the metabolite 4-methylumbelliferyl glucuronide (4-MUG), which makes it remarkable that 4-MU is effective at all. We report here that 4-MUG contributes to HA synthesis inhibition. We observed that oral administration of 4-MUG to mice inhibits HA synthesis, promotes FoxP3+ regulatory T-cell expansion, and prevents autoimmune diabetes. Mice fed either 4-MUG or 4-MU had equivalent 4-MU:4-MUG ratios in serum, liver and pancreas, indicating that 4-MU and 4-MUG reach an equilibrium in these tissues. LC-tandem MS experiments revealed that 4-MUG is hydrolyzed to 4-MU in serum, thereby greatly increasing the effective bioavailability of 4-MU. Moreover, using intra-vital 2-photon microscopy, we found that 4-MUG (a non-fluorescent molecule) undergoes conversion into 4-MU (a fluorescent molecule) and that 4-MU is extensively tissue bound in the liver, fat, muscle, and pancreas of treated mice. 4-MUG also suppressed HA synthesis independently of its conversion into 4-MU and without depletion of the HA precursor UDP-glucuronic acid (GlcUA). Together, these results indicate that 4-MUG both directly and indirectly inhibits HA synthesis and that the effective bioavailability of 4-MU is higher than previously thought. These findings greatly alter the experimental and therapeutic possibilities for HA synthesis inhibition.
View details for PubMedID 30914479
Donor-derived CIK Cell Infusion as Consolidation after Non-myeloablative Allogeneic Transplant for Myeloid Neoplasms.
Biology of blood and marrow transplantation : journal of the American Society for Blood and Marrow Transplantation
Non-myeloablative conditioning, such as with total lymphoid irradiation and anti-thymocyte globulin (TLI-ATG), has allowed hematopoietic allotransplantation with curative potential for older patients and those with comorbid medical conditions with myeloid neoplasms. However, early achievement of full donor chimerism (FDC) and relapse remain challenges. Cytokine induced killer (CIK) cells have been shown to have anti-tumor cytotoxicity. Infusion of donor-derived CIK cells has been studied for hematologic malignancies relapsed after allotransplant but has not been evaluated as post-transplant consolidation. In this phase II study, we prospectively studied whether a one-time infusion of 1?×?108/kg CD3+ donor-derived CIK cells administered between Days +21-35 after TLI-ATG conditioning, could improve FDC achievement by Day +90 and 2-year clinical outcomes in patients with myeloid neoplasms. CIK cells were infused in 31 of 44 patients treated on study and contained predominantly CD3+CD8+NKG2D+ cells along with significantly expanded CD3+CD56+ cells. Outcomes were compared to a retrospective historical cohort of 100 patients. We found that this one-time CIK infusion did not increase the rate of FDC by Day +90. On an intention-to-treat analysis, 2-year non-relapse mortality (6.8%, 95%CI: 0-14.5%), event-free survival (27.3%, 95%CI: 16.8-44.2%), and overall survival (50.6%, 95%CI: 37.5-68.2%) were similar to our historical cohort. Cumulative incidence of grade II-IV acute graft versus host disease at 1-year was 25.1% (95%CI: 12-38.2%). On univariate analysis, the presence of monosomal or complex karyotype was adversely associated with relapse-free and overall survival. Given the favorable safety profile of CIK cell infusion, strategies such as repeat dosing or genetic modification are worth exploration. This trial was registered at clinicaltrials.gov (NCT01392989).
View details for PubMedID 30951840