Discrimination of Treg subsets using Helios and FoxP3 followed by CDR3 sequencing reveals that the TCR repertoire of effector T Cells and natural regulatory T Cels is distinct

Wednesday, October 26, 2011 — Poster Session IV
2:00 p.m. – 4:00 p.m.	Natcher Conference Center	NIAID	IMMUNO/INFLAM-9

Authors

• A Golding
• F Hakim
• S Pavletic
• P Scheinberg
• D Douek
• J Melenhorst
• E Shevach

Abstract

The general conclusion of prior studies has been that the T cell receptor (TCR) repertoires of effector T cells (Teffs) and regulatory T cells (Tregs) overlap to some degree (10-20%). The prevailing model is that Tregs are selected during thymic development to recognize self-antigen whereas Teffs are selected primarily to express a repertoire that recognizes foreign antigen. In order to better identify Tregs, we have used a novel maker, Helios, together with FoxP3 to discriminate between thymically-derived natural Tregs (Helios positive) and peripherally-derived Tregs (Helios negative). We first show in individuals with skewed TCR VBeta repertoires, such as chronic GVHD patients and one patient with X-linked agammaglobulinemia, that Helios negative Tregs are over-represented by the VBeta that is over-expressed by Teffs. We have separately identified antigen-specific CD4 T cells using a Class II CMV Tetramer. We have gone on to sort Helios negative vs Helios positive Tregs vs Teffs and then used a DNA-based multiplex-PCR approach to sequence the VBeta TCR sequences of the three separate T lymphocyte groups. Our data shows that TCR CDR3 sequences are distinct and non-overlapping between Helios positive natural Tregs and FoxP3 negative Teffs.

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