Nat Methods. 2009 Jul;. Epub 2009 Jun 21

Abstract The use of fluorescently labeled MHC multimers has become an essential technique for the analysis of disease- and therapy-induced T cell immunity. While classical MHC multimer analyses are well-suited for the detection of immune responses against a few epitopes, limitations on patient sample size preclude a comprehensive analysis of T cell immunity. To address this issue, we have developed a combinatorial encoding strategy that allows the parallel detection of a multitude of different T cell populations within a single sample. Detection of antigen-specific T cells from peripheral blood by combinatorial encoding is as efficient as detection with conventional PE labeled multimers, but results in a significantly increased sensitivity, and most importantly, allows comprehensive screens to be performed on patient material. Proof of principle for the feasibility of large-scale screening of patient material was obtained by analysis of HLA-A3 restricted T cell responses against known and potential melanoma-associated antigens in peripheral blood from melanoma patients.