Detection of individual interleukin 2-producing cells after anti-CD3 antibody activation.
A new intracytoplasmic immunofluorescence staining technique to detect and quantify human interleukin-2 (IL-2)-producing lymphocytes is described. Blood mononuclear cells (MNC) were cultured and stimulated by mitogenic anti-CD3 antibody to produce IL-2. The cells were then fixed and subsequently permeabilized in suspension by the detergent saponin. Cytoplasmic IL-2 could then be demonstrated using polyclonal IL-2 specific antibodies and indirect immunofluorescence staining. A characteristic morphology of the IL-2 staining was noted with a local circle-shaped accumulation in the cytoplasm in a perinuclear position, probably reflecting the presence of the lymphokine in the Golgi stacks. A rapid, but transient IL-2 production peak 6 h after initiation of the cultures was observed. Only 1% of the cells produced IL-2, but each IL-2 stained cell was very bright, indicating a low capacity of anti-CD3 antibody to induce IL-2 production rather than an insensitivity of our detection system. Of the IL-2-producing cells 90% were CD4-positive T cells and 10% were CD8-positive T cells as revealed by two-colour staining of cell surface antigens and cytoplasmic IL-2.
Andersson
U
U
Department of Immunology, Stockholm University, Sweden.
Sander
B
B
D000911:Antibodies, Monoclonal; D000945:Antigens, Differentiation, T-Lymphocyte; D017252:CD3 Complex; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D005455:Fluorescent Antibody Technique; D006801:Humans; D066298:In Vitro Techniques; D007376:Interleukin-2 / Q000096:biosynthesis*; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000276:immunology*; D011948:Receptors, Antigen, T-Cell
D000911:Antibodies, Monoclonal; D000945:Antigens, Differentiation, T-Lymphocyte; D017252:CD3 Complex; D007376:Interleukin-2; D011948:Receptors, Antigen, T-Cell