A new approach to simultaneously quantify both TCR α-and β-chain diversity after adoptive immunotherapy

M Zhang, S Maiti, C Bernatchez, H Huls… - Clinical cancer …, 2012 - AACR
M Zhang, S Maiti, C Bernatchez, H Huls, B Rabinovich, RE Champlin, LM Vence, P Hwu
Clinical cancer research, 2012AACR
Purpose: T-cell receptor (TCR) variable Vα and Vβ gene diversity is a surrogate biomarker
for the therapeutic potential of adoptive immunotherapy and cellular immunity. Therefore,
creating a straightforward, rapid, sensitive, and reliable method to view the global changes
of both TCRVα and Vβ transcripts in heterogeneous populations of T cells is appealing.
Experimental Design: We designed a “direct TCR expression assay”(DTEA) using a panel of
customized bar-coded probes that simultaneously detects and quantifies 45 Vα and 46 Vβ …
Abstract
Purpose: T-cell receptor (TCR) variable Vα and Vβ gene diversity is a surrogate biomarker for the therapeutic potential of adoptive immunotherapy and cellular immunity. Therefore, creating a straightforward, rapid, sensitive, and reliable method to view the global changes of both TCRVα and Vβ transcripts in heterogeneous populations of T cells is appealing.
Experimental Design: We designed a “direct TCR expression assay” (DTEA) using a panel of customized bar-coded probes that simultaneously detects and quantifies 45 Vα and 46 Vβ transcripts in a nonenzymatic digital multiplexed assay from a small number of cells (104 cells) or as little as 100 ng of total RNA.
Results: We evaluated DTEA on total RNA samples of tumor-infiltrating lymphocytes and peripheral blood obtained from patients with melanoma after adoptive T-cell therapy. DTEA detected a similar spectrum of the dominant patterns of TCRVβ gene usage as sequencing cloned TCRVβ CDR3 regions. However, DTEA was rapid, achieved a level of sensitivity to identify rare T-cell populations, and simultaneously tracked the full array of Vα and Vβ transcripts.
Conclusions: DTEA can rapidly and sensitively track changes in TCRVα and gene usages in T-cell pools following immune interventions, such as adoptive T-cell transfer, and may also be used to assess impact of vaccination or reconstitution of T-cell compartment after hematopoietic stem cell transplantation. Clin Cancer Res; 18(17); 4733–42. ©2012 AACR.
AACR