title: Unbiased molecular analysis of T cell receptor expression using template-switch anchored RT-PCR.
abstract: A detailed knowledge of the principles that guide clonal selection within the memory and effector T cell pools is essential to further our understanding of the factors that influence effective T cell-mediated immunity and has direct implications for the rational design of vaccines and immunotherapies. This unit provides methods for the unbiased quantification and characterization of all expressed T cell receptor (TCR) gene products within any defined T cell population. The approach is based on a template-switch anchored reverse transcription-polymerase chain reaction (RT-PCR) and is optimized for the analysis of antigen-specific T cells isolated directly ex vivo.
authors.affiliation: Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Máire F
authors.initials: MF
authors.lastname: Quigley
authors.forename: Jorge R
authors.initials: JR
authors.lastname: Almeida
authors.forename: David A
authors.initials: DA
authors.lastname: Price
authors.forename: Daniel C
authors.initials: DC
authors.lastname: Douek
mesh_terms: D000818:Animals; D002999:Clone Cells; D019672:Genes, T-Cell Receptor / Q000235:genetics; D006801:Humans; D007111:Immunity, Cellular / Q000235:genetics; D057089:Pathology, Molecular / Q000295:instrumentation / Q000379:methods; D016133:Polymerase Chain Reaction; D037182:T-Cell Antigen Receptor Specificity; D013601:T-Lymphocytes / Q000276:immunology*; D013698:Templates, Genetic