Single cell RNA seq profiling of (auto-)antigen-specific CD8+ T cells in response to their target antigen

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Abstract Summary

 

CD8+ T cells are key player of the adaptive immune response. Improving our understanding of antigen-specific T cell responses on the single cell level will be crucial for the development of highly effective future vaccines and a necessity to resolve or minimize errant immunity in autoimmune diseases and after transplantation. Currently, the profiling of peptide-specific CD8+ T cells is compromised by a low efficiency of multimer-based isolation following peptide stimulation.

 

We established an assay that circumvents the isolation of antigen-responsive CD8+ T cells via MHC class I multimers and applied it to the analysis of CD8+ T cells responding to their cognate viral antigen (Influenza M158-66, CMV pp65495-503), beta cell autoantigen (IGRP265-273) or control stimuli. Targeted single cell gene expression analysis and single cell RNA sequencing on more than 600 isolated cells was conducted to identify genes differentially expressed in response to the cognate antigen.

 

We identified sets of transcripts that clearly mark the antiviral response on a single cell level, across donors and that largely overlap for cells stimulated via PBMC or cell line-based antigen presentation. The approach also identified paired TCR alpha and beta chain sequences with the expected TCR gene usage for flu and CMV responses. Fewer differentially expressed genes were identified for IGRP-responsive cells and these differed from those seen in antiviral response. Again, paired TCR alpha and beta chain sequences could be obtained from the IGRP-responsive cells.

 

A valid method for the isolation of peptide responsive CD8+ T cells was developed. Cells isolated using this method provide detailed information on the response profile and TCR, and suggest that circulating beta cell antigen peptide-stimulated CD8+ T cells have a different gene expression profile than viral peptide-responsive CD8+ T cells.

 

Submission ID :
IDS36218
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DFG-Center for Regenerative Therapies Dresden, Faculty of Medicine, Technische Universität Dresden, Dresden, Germany
CRTD-DFG Center for Regenerative Therapies Dresden, Carl Gustav Carus Faculty of Medicine, Technische Universität Dresden, Dresden, Germany
CRTD Center for Regenerative Therapies Dresden
DFG-Center for Regenerative Therapies Dresden, Faculty of Medicine, Technische Universität Dresden, Dresden, Germany
DFG-Center for Regenerative Therapies Dresden, Faculty of Medicine, Technische Universität Dresden, Dresden, Germany
DFG-Center for Regenerative Therapies Dresden, Faculty of Medicine, Technische Universität Dresden, Dresden, Germany
Institute of Diabetes Research, Helmholtz Zentrum München, German Research Center for Environmental Health, Munich-Neuherberg, Germany
CRTD Center for Regenerative Therapies Dresden

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KEY DATES

Event dates:
Thursday 25 October - Monday 29 October 2018

Abstract submission deadline:
Monday 14 May 2018

Abstract notification:
July 2018

Early registration deadline:
Monday 3 September 2018

Registration deadline:
Monday 15 October 2018

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British Society for Immunology
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