# TCR Sequencing Core

> **NIH NIH U19** · COLUMBIA UNIVERSITY HEALTH SCIENCES · 2020 · $361,134

## Abstract

Summary 
The overall goal of the U19 proposal is to combine kidney and islet transplantation with nonmyeloablative 
hematopoietic cell transplantation (HCT) to achieve immune tolerance to the donor tissues. We will use a the 
cynomolgus monkey model to develop this approach for deceased and living donor scenarios, using expanded 
polyclonal and donor-specific recipient regulatory T cells respectively, in Projects 1 and 2. We hypothesize that 
tolerance will involve both expansion and induction of donor-specific regulatory T cells and deletion of donor- 
specific effector T cells. A key to development of this procedure will be the tools to identify and track these 
donor-specific T cells. We previously developed such a method in humans. Core B will extend this technology 
to cynomolgus monkeys (cyno) and apply the assay to support Projects 1 and 2. Specifically, Core B will follow 
the same development pipeline we have used for human and mouse to develop a T cell receptor (TCR) 
sequencing assay for cyno. Since TCRs rearrange somatically with massive diversity, each TCR is nearly 
unique. This allows us to track hundreds of thousands of T cell clones over time and between tissues by their 
TCR sequence. We leverage with technology by first isolating donor-specific T cells through a mixed lymphocyte 
reaction (explicitly stimulating T cells that are specific to the donor tissue). The clones in these expanded donor- 
reactive CD4 and CD8 T cells are compared to those in sorted, unstimulated functional subsets, including effector 
and regulatory T cells, and the expanded sequences are identified as belonging to the donor-reactive subset. 
We will validate this assay in cynomolgus monkeys receiving allotransplants and optimize methods .for defining 
donor-reactive effector and regulatory T cell clones. Timed, sorted T cell DNA samples from infused Tregs, 
peripheral blood and graft-infiltrating lymphocytes as well as DNA from biopsy specimens and urine pellets from 
transplanted cyno monkeys in Projects 1 and 2 will be sent to Core B, where the assay will be applied. Over 
time, we will monitor the clonal expansion and contraction of these T cell subsets in blood, tissue, and urine 
pellets to identify mechanisms of immune tolerance in this preclinical model.

## Key facts

- **NIH application ID:** 9988360
- **Project number:** 5U19AI131474-04
- **Recipient organization:** COLUMBIA UNIVERSITY HEALTH SCIENCES
- **Principal Investigator:** Jason H Bielas
- **Activity code:** U19 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $361,134
- **Award type:** 5
- **Project period:** 2017-08-18 → 2022-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9988360

## Citation

> US National Institutes of Health, RePORTER application 9988360, TCR Sequencing Core (5U19AI131474-04). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/9988360. Licensed CC0.

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