A new approach to simultaneously quantify both TCR ?- and ?-chain diversity after adoptive immunotherapy.

ABSTRACT: T-cell receptor (TCR) variable V? and V? 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 TCRV? and V? transcripts in heterogeneous populations of T cells is appealing.We designed a "direct TCR expression assay" (DTEA) using a panel of customized bar-coded probes that simultaneously detects and quantifies 45 V? and 46 V? transcripts in a nonenzymatic digital multiplexed assay from a small number of cells (10(4) cells) or as little as 100 ng of total RNA.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 TCRV? gene usage as sequencing cloned TCRV? CDR3 regions. However, DTEA was rapid, achieved a level of sensitivity to identify rare T-cell populations, and simultaneously tracked the full array of V? and V? transcripts.DTEA can rapidly and sensitively track changes in TCRV? and V? 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.

SUBMITTER: Zhang M 

PROVIDER: S-EPMC3823368 | biostudies-literature | 2012 Sep

REPOSITORIES: biostudies-literature