Detection of intracellular expression and secretion of interferon-gamma at the single-cell level after activation of human T cells with tetanus toxoid in vitro.
Activation of T cells results in intracellular expression and secretion of cytokines such as interferon (IFN)-gamma. Here we have used three different assays for determination of IFN-gamma in tetanus toxoid- or mitogen-activated human T cell cultures. Two of these assays [intracytoplasmic immunofluorescence and enzyme-linked immuno spot assay (ELISPOT)] determined the expression and secretion of IFN-gamma at the single-cell level while the third assay enzyme-linked immunosorbent assay (ELISA) measured IFN-gamma secreted into the culture supernatant. Comparison of all three tests revealed a good correlation between the ELISPOT assay and the ELISA, whereas expression of intracellular IFN-gamma showed a qualitative but not a quantitative correlation with the latter. Both the immunospot assay and the immunofluorescence may be used to detect approximate numbers of specific T cells even when present at low frequencies. With the use of the immunospot assay antigen-specific T cells could be detected even in the absence of detectable IFN-gamma in the culture supernatants. However, the ELISA assay should be more convenient for screening large clinical material.
Kabilan
L
L
Department of Immunology, University of Stockholm, Sweden.
Andersson
G
G
Lolli
F
F
Ekre
H P
HP
Olsson
T
T
Troye-Blomberg
M
M
D000941:Antigens / Q000276:immunology; D002478:Cells, Cultured; D003470:Culture Media / Q000032:analysis; D003593:Cytoplasm / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D006801:Humans; D007158:Immunologic Techniques; D007371:Interferon-gamma / Q000096:biosynthesis* / Q000378:metabolism; D008213:Lymphocyte Activation / Q000276:immunology; D010835:Phytohemagglutinins; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism*; D013745:Tetanus Toxoid / Q000276:immunology
D000941:Antigens; D003470:Culture Media; D010835:Phytohemagglutinins; D013745:Tetanus Toxoid; D007371:Interferon-gamma