Detection of soluble T cell receptor-releasing cells by ELISPOT assay

A specific and sensitive enzyme-linked immunospot (ELISPOT) assay has been developed for the detection and enumeration of soluble T cell receptor (TCR)-releasing cells. Using this method, we readily detected at the single cell level the release of soluble TCR by living T lymphoma cells (MT-2 and HSB-2) but not by human B lymphoma cells (DAKIKI), mouse hepatoma cells (MH134) and dead MT-2. Furthermore, distinct spots in MT-2 cell culture were not visualized using several monoclonal antibodies against antigens unrelated to TCRs as a primary antibody. The specific and quantitative detection of soluble TCR-releasing cells using ELISPOT assay will certainly provide a valuable tool to better characterize soluble TCRs and their relationship to immune regulation and a number of diseases.