Saturday, February 10, 2018

Antigen-specific Foxp3+ Tregs control tolerance to gut pathobiont H. hepaticus

Helicobacter hepaticusH. hepaticus, is an opportunist commensal, a pathobiont, that causes gut inflammation in IL-10 deficient but not in WT mice. A New study in journal Nature from Dan Littman's lab tried to understand basis for such dual nature of host response to H. hepaticus.

The authors have generated transgenic T cells specific for H. hepaticus-unique protein, HH_1713. Next, the authors co-transferred H. hepaticus-specific T cells (HH7-2tg cells) alongside with segmented filamentous bacteriaSFB-specific T cells (7B8tg cells) into WT mice exposed to H. hepaticus. Interestingly, HH7-2tg cells mostly differentiated into RORγt+ Foxp3+ Tregs while 7B8tg cells mostly differentiated into Th17 cells, as observed in earlier studies.

However, similar transfer of H. hepaticus-specific T cells into IL-10KO mice exposed to H. hepaticus mostly yielded inflammatory T cells (Th17 and Th1 cells).

It appears that transcription factor c-MAF control differentiation of H. hepaticus-specific T cells into RORγt+ Foxp3+ Tregs and its deficiency recapitulates H. hepaticus-specific T cells differentiation into inflammatory T helper cells observed in IL-10KO hosts when exposed to H. hepaticus.

In summary, this study indicates that in WT mice harbor H. hepaticus specific Foxp3+ T cells that keep tolerance to H. hepaticus. These induced Foxp3+ T cells are probably generated (or expand) by chain reaction orchestrated by existed antigen-specific natural Tregs since induced Tregs don't develop efficiently when transferred into RAG KO hosts lacking natural Tregs. IL-10 or c-MAF deficiency could alter overall microbiota composition (unrelated to H. hepaticus) and drive shift from Treg program into inflammatory pathway.

posted by David Usharauli

CD4 T cells silence innate over-activation to gut microbiota

A simple but very effective study was published in journal Nature from Ron Germain's lab. His group is known for publishing high quality in situ microscopy data combined with cellular analysis. New study continues this trend. 

In this study the authors analyzed pattern of phosphorylation of STAT3 transcription factor in small intestine derived from various immune deficient mouse strains. Compared to WT mice, pSTAT3 staining in RAG1 KO gut tissue (lacking adaptive immune system) was significantly up-regulated. 

Analysis of antibiotic-treated or germ-free mice indicated that pattern of pSTAT3 staining in RAG1-KO was correlated with the presence of gut microbiota.

Interestingly, longitudinal analysis showed that pSTAT3 staining inversely correlated with  maturation of adaptive immune system post weaning (between 4-20 weeks). 

Co-housing experiments showed that T cells, but not B cells, played a role in silencing innate pSTAT3 over-activation.

And out of T cells, it were CD4 T cells and class II antigen-presentation that played the role in pSTAT3 silencing.

Finally, both Tregs and SBF-specific Th17 cells (7B8 transgenic T cells) could mediate silencing of pSTAT3 over-activation. Both T cell type could down-regulate STAT3 phosphorylation in innate and epithelial cells but the mechanisms could be different. 

In summary, this study showed that persistent pSTAT3 over-activation observed in mice deficient for CD4 T cell function could explain some of chronic metabolic shifts observed in clinical settings. 

posted by David Usharauli

Tuesday, February 6, 2018

T1D target epitope from zinc transporter 8 (ZnT8) cross-reacts with commensal bacteria

Type 1 diabetes (T1D) is considered autoimmune disease. Of course, in humans, we don't have a direct evidence that islet-specific auto-reactive T cells and auto-antibodies found in peripheral blood from T1D patients are indeed responsible for tissue damage. Such evidence would require T/B cell and Ab depletion experiment that is not feasible. Autoimmune nature of T1D is basically extrapolated from mouse studies or in vitro antigen binding assays.   

Several islet antigens are known to represent targets in T1D, such as preproinsulin (PPI), glutamic acid decarboxylase (GAD), insulinoma-associated protein-2 (IA-2), islet-specific glucose-6-phosphatase catalytic subunit-related protein (IGRP) and more recently described zinc transporter 8 (its peptide ZnT8186–194).

New study in Science Immunology found that ZnT8186–194-specific CD8 T cells are largely similarly present in both T1D patients and healthy controls and that ZnT8186–194-specific CD8 T cells could recognize (cross-react) peptide derived from gut commensal microbe Bacteroides stercoris.

Staining with ZnT8186–194-specific HLA class I multimers (MMr) and other functional antigen-specific assays found that T1D and healthy controls harbored largely similar number and functional ZnT8186–194-specific CD8 T cells.

Interestingly, ZnT8186–194-specific CD8 T cells could be double stained with HLA class I multimer + peptide derived from B. stercoris, a commensal bacterial species found in gut flora. It is not the first time such cross-reactivity has been observed between islet-specific CD8 T cells and commensal bacteria. Previously, at least two bacterial species have been identified to cross-react with IGRP-specific CD8 T cells.

In summary, this study indicates that deletion of auto-reactive CD8 T cells in the thymus is not sufficient to prevent autoimmunity and that regulatory mechanisms operating in the periphery is necessary to prevent initiation of auto-reactive attack by circulating ZnT8186–194-specific CD8 T cells (probably Tregs the authors had in mind). Cross-reactive peptide derived from B. stercoris could play role in priming of ZnT8186–194-specific CD8 T cells in absence of regulatory circuit. But how and why such antigen-specific tolerance breakdown happens in one and not in another is not clear at this stage.

posted by David Usharauli