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

Clin Cancer Res. 2012 Sep 1;18(17):4733-42. doi: 10.1158/1078-0432.CCR-11-3234. Epub 2012 Jul 3.


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 (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 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 Vβ 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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers, Pharmacological / blood
  • Humans
  • Immunotherapy, Adoptive*
  • Lymphocytes, Tumor-Infiltrating / metabolism
  • Melanoma / blood*
  • Melanoma / immunology
  • Melanoma / therapy
  • RNA / blood
  • RNA / genetics
  • Receptors, Antigen, T-Cell, alpha-beta / blood
  • Receptors, Antigen, T-Cell, alpha-beta / genetics*


  • Biomarkers, Pharmacological
  • Receptors, Antigen, T-Cell, alpha-beta
  • RNA