# In vivo development and reactivity of human autoreactive T cells

> **NIH NIH R01** · COLUMBIA UNIVERSITY HEALTH SCIENCES · 2021 · $405,000

## Abstract

Abstract
Inadequate recognition of autoantigens during thymopoiesis can result in abnormally high frequency of
autoreactive T cells, which can lead to autoimmune diseases when combined with defective mechanisms of
peripheral tolerance in susceptible individuals. The process of thymic selection of autoreactive T cells in humans
is largely not understood due to lack of in vivo models and could so far only be extrapolated from rodent studies.
Analysis of autoantigen expression in the human thymus has provided an incomplete picture based on increasing
evidence that this does not necessarily correlate with T cell deletion. As autoreactive T cells are also found
circulating in `healthy' individuals, it is unclear to what extent incomplete thymic deletion (or regulatory T cell
(Treg) selection) contributes to autoimmunity.
Our proposed research is based on humanized mice reconstituted with fetal hematopoietic stem cells (HSCs)
and thymic tissue, whereby HSCs can be transduced to express an autoreactive T cell receptor (TCR) and/or
the actual epitope recognized by this TCR. The HSCs then develop into T cells in a normal human thymus in
vivo, a fraction of which display the autoreactive TCR. Having established the model with T cells specific for a
melanocyte autoantigen that fail to undergo thymic deletion unless the epitope is also introduced in HSCs, we
will adapt this system to a collection of TCRs that have been identified in islet infiltrates of Type 1 diabetes
patients. These TCRs were selected because it is yet unknown whether their specific epitopes are presented
and recognized in a normal human thymus. Thus the model will help determine whether T cells bearing those
TCRs escape thymic deletion through a number of mechanisms that will be tested (lack of expression, of cross-
reactivity or of neoepitope formation). Introduction of the cognate epitopes in HSCs will allow us to evaluate their
effect on developing autoreactive T cells (deletion versus positive selection of Tregs), and further explore the
unique contribution of specific subsets of antigen-presenting cells (APCs) in this process, including their ability
to relocate to the thymus from the bloodstream to influence thymic selection. Using recipient mice that are able
to present antigens to human T cells and antigens that are cross-reactive between the two species, we will
assess the conditions that are needed to activate autoreactive T cells to attack their target tissue.
This work will feature immunofluorescence imaging, multi-parametric flow cytometry and single-cell RNA
sequencing analysis to analyze human APC subsets that populate the thymus and peripheral lymphoid tissues
and engage human autoreactive T cells, and to assess the phenotype and molecular profile of these autoreactive
T cells based on different conditions of antigen encounter (thymus vs periphery, type of APC, etc). Overall, these
studies will provide valuable insights into the generation of human autoreactive T cells cau...

## Key facts

- **NIH application ID:** 10054162
- **Project number:** 5R01AI142428-03
- **Recipient organization:** COLUMBIA UNIVERSITY HEALTH SCIENCES
- **Principal Investigator:** Remi J Creusot
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $405,000
- **Award type:** 5
- **Project period:** 2018-11-15 → 2023-10-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10054162

## Citation

> US National Institutes of Health, RePORTER application 10054162, In vivo development and reactivity of human autoreactive T cells (5R01AI142428-03). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10054162. Licensed CC0.

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