Evaluation of a single-platform, MHC tetramer based, six-parameter flow cytometric method for immunmonitoring

A major setback in tumor vaccine therapy is the lack of an adaequate and easy to use immune monitoring system. The quantitation of antigen-specific T-cells would provide an insight into the development and dynamics of the induced T-cell responses. In order to establish the monitoring system, we used pp65, immundominant epitope of CMV as a model antigen. Whole blood from healthy donors was stained with HLA-A*0201/tetramers that stain the pp65 specific T-cell receptor. A single-platform, four-color flow cytometric assay was established to obtain absolute counts of tetramer-positive cells. Various staining and gati…