Solid-phase selection of human T lymphocyte subpopulations using monoclonal antibodies.
This report describes a novel solid-phase technique for the positive selection of human T lymphocyte subsets labeled by indirect immunofluorescence with monoclonal anti-bodies. Fluorescein labeled normal human T cells or a T cell line were fractionated on plastic culture dishes precoated with affinity chromatography purified anti-fluorescein antibodies. Cell binding was specific for fluorescein, and was both time and temperature dependent. Bound cells were eluted at 37 degrees C with fluorescein-L-lysine. The eluted cells were enriched with highly viable and functional human T cell subsets. Thus Leu3 monoclonal antibody selected cells were shown to provide helper activity in the pokeweed mitogen induced IgM and IgG immunoglobulin secretory response of autologous B cells. The Leu2 antibody selected T cells suppressed both IgM and IgG secretory responses. In addition, studies with the monoclonal antibody 1G11, which binds to an antigen expressed on acute lymphocytic leukemia (T-ALL) cells and the T-ALL derived cell line RPMI-8402, demonstrated that this solid-phase technique can be used to select for cells which are present at low frequencies in a mixed population. It thus provides a simple and reproducible means for the preparative isolation of lymphocyte subsets associated with autoimmune and neoplastic disease for functional and biochemical analysis.
D000818:Animals; D000911:Antibodies, Monoclonal; D002469:Cell Separation; D002478:Cells, Cultured; D005452:Fluoresceins; D005455:Fluorescent Antibody Technique; D006801:Humans; D007945:Leukemia, Lymphoid / Q000276:immunology; D051379:Mice; D008815:Mice, Inbred Strains; D011817:Rabbits; D013601:T-Lymphocytes / Q000145:classification*
D000911:Antibodies, Monoclonal; D005452:Fluoresceins