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. Curr. Protoc. Immunol. 94:10.33.1‐10.33.16. © 2011 by John Wiley & Sons, Inc.