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.
Ishizaka
S
S
Department of Parasitology, Nara Medical University, Japan.
Kimoto
M
M
Nishiyama
T
T
Araki
T
T
D017077:Culture Media, Conditioned; D003513:Cycloheximide / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D016393:Lymphoma, B-Cell / Q000276:immunology; D016399:Lymphoma, T-Cell / Q000276:immunology; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000378:metabolism*; D013601:T-Lymphocytes / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D017077:Culture Media, Conditioned; D016693:Receptors, Antigen, T-Cell, alpha-beta; D003513:Cycloheximide