title: A new approach to simultaneously quantify both TCR α- and β-chain diversity after adoptive immunotherapy.
abstract: OBJECTIVE
  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.
  
  
  METHODS
  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.
authors: Department of Melanoma Medical Oncology, Unit 904, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Blvd., Houston, TX 77030, USA.
authors: Minying
authors: M
authors: Zhang
authors: Sourindra
authors: S
authors: Maiti
authors: Chantale
authors: C
authors: Bernatchez
authors: Helen
authors: H
authors: Huls
authors: Brian
authors: B
authors: Rabinovich
authors: Richard E
authors: RE
authors: Champlin
authors: Luis M
authors: LM
authors: Vence
authors: Patrick
authors: P
authors: Hwu
authors: Laszlo
authors: L
authors: Radvanyi
authors: Laurence J N
authors: LJ
authors: Cooper
mesh_terms: D054316:Biomarkers, Pharmacological / Q000097:blood; D006801:Humans; D016219:Immunotherapy, Adoptive; D016246:Lymphocytes, Tumor-Infiltrating / Q000378:metabolism; D008545:Melanoma / Q000097:blood* / Q000276:immunology / Q000628:therapy; D012313:RNA / Q000097:blood / Q000235:genetics; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000097:blood / Q000235:genetics*
chemical_list: D054316:Biomarkers, Pharmacological; D016693:Receptors, Antigen, T-Cell, alpha-beta; D012313:RNA