Solid-phase selection of human T lymphocyte subpopulations using monoclonal antibodies

Abstract

This report describes a novel solid-phase technique for the positive selection of human T lymphocyte subsets labeled by indirect immunofluorescence with monoclonal antibodies. 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°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 bonds 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.