Characterization of individual tumor necrosis factor alpha-and beta-producing cells after polyclonal T cell activation.
Mononuclear cells from human blood were stimulated to tumor necrosis factor alpha (TNF alpha) or beta (TNF beta) production by the T cell mitogens anti-CD3 antibody (OKT3) or staphylococcal enterotoxin A (SEA). The cells were then fixed and subsequently permeabilized in suspension by the detergent saponin in order to enable TNF alpha- or TNF beta-specific antibodies to enter the cells and interact with cytoplasmic TNF in producer cells. A characteristic morphology of the staining pattern of the two cytokines was noted, with a local accumulation in the cytoplasm in a perinuclear position reflecting the presence of TNF alpha or -beta in the Golgi system. TNF alpha-producing cells appeared 2-3 h after activation of the cultures and increased up to 6 h. The majority of these early TNF alpha-producing cells were monocytes as judged by two-color staining and morphology, but a small fraction of CD4- and CD8-positive T cells was found up to 72 h. TNF beta production started later and peaked 18 or 48 h after OKT3 or SEA stimulation, respectively. The number of TNF beta-producing cells was much larger than that of TNF alpha-producing cells, and approximately 90% of them were CD4-positive T cells. The remaining TNF beta production occurred in CD8-positive T cells and in B cells. Almost every second CD4-positive T cell made TNF beta at the peak of the SEA-induced synthesis. The cytotoxic activity found in the supernatants correlated well with the number of TNF-producing cells found in the cultures. Cells from fresh blood or unstimulated cultures showed no or very few TNF-producing cells.
Andersson
U
U
Department of Immunology, Stockholm University, Arrhenius Laboratories for Natural Sciences, Sweden.
Adolf
G
G
Dohlsten
M
M
Möller
G
G
Sjögren
H O
HO
D015703:Antigens, CD / Q000032:analysis; D000945:Antigens, Differentiation, T-Lymphocyte / Q000502:physiology; D017252:CD3 Complex; D002478:Cells, Cultured; D004768:Enterotoxins / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D006684:HLA-DR Antigens / Q000032:analysis; D006801:Humans; D066298:In Vitro Techniques; D008213:Lymphocyte Activation; D008233:Lymphotoxin-alpha / Q000096:biosynthesis*; D011948:Receptors, Antigen, T-Cell / Q000502:physiology; D013601:T-Lymphocytes / Q000502:physiology*; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis*
D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D017252:CD3 Complex; D004768:Enterotoxins; D006684:HLA-DR Antigens; D008233:Lymphotoxin-alpha; D011948:Receptors, Antigen, T-Cell; D014409:Tumor Necrosis Factor-alpha