Rapid separation of T cell subpopulations with monoclonal antibodies and affinity chromatography.
A rapid method for isolation of highly enriched helper and suppressor T cell subsets and their corresponding helper and suppressor cell depleted cell populations is described. The method is based on the binding of monoclonal antibodies to helper and suppressor cells and subsequent affinity chromatography with covalently bound rabbit anti-mouse antibodies. As assessed by indirect immunofluorescence, purity of the enriched subpopulations exceeds 90%, whereas no contamination with helper or suppressor cells is detectable in populations depleted of the respective subsets. The cells isolated by this method show no functional defects in helper and suppressor assays and respond with increased DNA synthesis to stimulation with phytohemagglutinin (PHA) plus 2-mercaptoethanol (2-ME).
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D002448:Cell Adhesion; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D002499:Centrifugation, Density Gradient; D002846:Chromatography, Affinity; D005455:Fluorescent Antibody Technique; D006801:Humans; D008213:Lymphocyte Activation; D051379:Mice; D010835:Phytohemagglutinins / Q000494:pharmacology; D011817:Rabbits; D013601:T-Lymphocytes / Q000145:classification* / Q000276:immunology; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology; D050378:T-Lymphocytes, Regulatory / Q000276:immunology
D000911:Antibodies, Monoclonal; D010835:Phytohemagglutinins