Human interleukin 6 and tumor necrosis factor alpha production studied at a single-cell level.
Individual peripheral blood mononuclear cells, which produced interleukin 6 (IL 6) or tumor necrosis factor alpha, (TNF alpha), were studied by cytokine-specific polyclonal or monoclonal antibodies (mAb) and immunofluorescence technique with UV microscopy. Lipopolysaccharide (LPS) induced IL 6 as well as TNF alpha production in the majority of the monocytes, but not at all in lymphocytes. Approximately every second monocyte made TNF alpha in response to LPS within 0.5 h from start of the cultures, when no IL 6 or TNF alpha production occurred. The maximal number of TNF alpha-synthesizing monocytes was observed 1.5 h later and then rapidly declined. LPS stimulation led to optimal IL 6 production 3 h after initiation of the cultures, with 90% of the monocytes expressing intracellular IL 6. LPS-induced IL 6 synthesis started about 1 h after that of TNF alpha. Polyclonal T cell activation with staphylococcal enterotoxin A or anti-CD3 mAb induced a biphasic production pattern of IL 6 as well as TNF alpha. Early IL 6 synthesis, which peaked 6-8 h from start of the cultures, occurred exclusively in monocytes, while late IL 6 production at 48 h was restricted to a small fraction of lymphoid cells. T cell mitogen induced early TNF alpha production, which peaked at 6 h, took mainly place in monocytes and to a minor degree in CD4+ as well as CD8+ T cells. The majority of the TNF alpha-producing mononuclear cells at 24 h were of the CD4+ T cell lineage in the staphylococcal enterotoxin A- or anti-CD3 mAb-activated cultures. IL 6 as well as TNF alpha accumulated in the Golgi system, which resulted in a characteristic morphology of the staining, eliminating problems with evaluation of background signals.
Andersson
U
U
Department of Immunology, Stockholm University, Sweden.
Matsuda
T
T
D000945:Antigens, Differentiation, T-Lymphocyte / Q000502:physiology; D017252:CD3 Complex; D002478:Cells, Cultured; D003593:Cytoplasm / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006056:Golgi Apparatus / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D015850:Interleukin-6; D007378:Interleukins / Q000096:biosynthesis*; D007963:Leukocytes, Mononuclear / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000494:pharmacology; D011948:Receptors, Antigen, T-Cell / Q000502:physiology; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis*
D000945:Antigens, Differentiation, T-Lymphocyte; D017252:CD3 Complex; D015850:Interleukin-6; D007378:Interleukins; D008070:Lipopolysaccharides; D011948:Receptors, Antigen, T-Cell; D014409:Tumor Necrosis Factor-alpha