Determination of the T3-3A1 antigen in PHA-induced human T-cells by standardized cell-ELISA.
The treatment of mononuclear leukocytes (MNL) with lectins induces marked changes in the cell's morphology, physiology and the composition of the cell surface. We used an immunoassay to monitor the PHA-induced expression of the T-lymphocyte-specific antigen T3-3A1 in fixed MNL with a monoclonal antibody (MoAb) specific for this antigen. This assay permits the detection and quantitation of the T3-3A1 antigen in a few thousand cells without the use of a FACS. The test was calibrated with isolated plasma membranes and, combined with a total protein determination, the relative content of T3-3A1 antigen in each sample could be calculated. Maximal T3-3A1 synthesis required a 10-fold lower concentration of PHA than was necessary for optimal DNA synthesis. The test may be used to screen for PHA stimulation.
D000911:Antibodies, Monoclonal / Q000276:immunology; D000941:Antigens / Q000032:analysis* / Q000276:immunology; D002462:Cell Membrane / Q000276:immunology; D004261:DNA Replication; D004306:Dose-Response Relationship, Immunologic; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D008213:Lymphocyte Activation / Q000187:drug effects; D010835:Phytohemagglutinins / Q000494:pharmacology; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000941:Antigens; D010835:Phytohemagglutinins; C051383:T3-3A1 antigen