A standard microcytotoxicity technique for quantitative analysis of lymphocyte subsets. A comparison with indirect immunofluorescence, evaluated by microscopy or flow cytometry.
A standard complement-dependent microcytotoxicity (CDC) technique was used for quantitative analysis of T-lymphocyte subsets in human peripheral blood and the results compared to those obtained by indirect immunofluorescence microscopy and flow cytometry. The monoclonal antibodies OKT3, OKT4 and OKT8 were used in the CDC method for detection of total-T cells, T-helper and T-suppressor cells respectively. The CDC technique provided reproducible results (CV, 3-7%) correlating well with both immunofluorescence techniques. This observation was valid both for healthy persons (n = 21) and for patients (n = 10) with immunological disorders. The correct antibody dilution, correction for background and the use of eosin staining are considered critical for the usefulness of this technique. The method has several advantages: it is widely used for histocompatibility testing, only simple equipment is necessary, and the amount of monoclonal antibody required per test is small.
D000911:Antibodies, Monoclonal; D003165:Complement System Proteins / Q000502:physiology; D003601:Cytotoxicity Tests, Immunologic / Q000379:methods*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D007958:Leukocyte Count; D008856:Microscopy, Fluorescence; D013601:T-Lymphocytes / Q000145:classification*
D000911:Antibodies, Monoclonal; D003165:Complement System Proteins