Journal
CLINICAL CANCER RESEARCH
Volume 18, Issue 17, Pages 4733-4742Publisher
AMER ASSOC CANCER RESEARCH
DOI: 10.1158/1078-0432.CCR-11-3234
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Funding
- NIH [CA111999, CA16672, CA124782, CA120956, CA141303, CA116127, 1S10RR026916]
- Burroughs Wellcome Fund
- Cancer Prevention Research Institute of Texas
- CLL Global Research Foundation
- Department of Defense
- Dr. Miriam and Sheldon Adelson Medical Research Foundation
- Estate of Noelan L. Bibler
- Harry T. Mangurian, Jr., Fund for Leukemia Immunotherapy
- Gillson Longenbaugh Foundation
- Institute of Personalized Cancer Therapy
- Melanoma Research Alliance
- Miller Foundation
- Mr. and Mrs. Joe H. Scales
- National Foundation for Cancer Research
- Pediatric Cancer Research Foundation
- Sister Institution Network Fund
- William Lawrence and Blanche Hughes Children's Foundation
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Purpose: T-cell receptor (TCR) variable V alpha and V beta 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 alpha and V beta 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 V alpha and 46 V beta 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. 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 TCRV beta gene usage as sequencing cloned TCRV beta 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 alpha and V beta transcripts. Conclusions: DTEA can rapidly and sensitively track changes in TCRV alpha and V beta 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. (C) 2012 AACR.
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