A novel approach to priming and assessing the diversity of T-cell receptors using randomized peptide pools

Thursday, October 11, 2012 — Poster Session IV
2:00 p.m. – 4:00 p.m.	Natcher Conference Center, Building 45	NIAID	IMMUNO-5

Authors

• J.S. Barber
• L. Yokomizo
• J.D. Milner

Abstract

Functional evaluation of polyclonal T-cell receptor (TCR) repertoires is difficult due to the large number of unique TCRs, a feature that also makes it difficult to activate polyclonal T-cell populations with their most physiologic agonist—peptides. Peripheral blood mononuclear cells (PBMC), or sorted naïve CD4 T-cells with autologous DCs were mixed with pools of 15-mer peptides comprised of a single fixed amino acid with the other 14 randomized. Across numerous donors, peptide pools led to a range of activation and proliferation of CD4 cells from PBMC, and to differentiation and Thelper polarization of naïve cells. This technique permitted discrimination of TCR diversity in several models of human disease associated with limited repertoire and allowed for evaluation of functional overlap between effector and Tregulatory cells. Randomized peptide pools can be used to assess diversity in patients with suspected reductions in TCR repertoire and to physiologically study activation and differentiation of naïve human T-cells.

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