title: Binding of human TNF-alpha to high-affinity cell surface receptors: effect of IFN.
abstract: Human tumor-necrosis factor (INF-alpha), induced by Sendai virus in peripheral blood mononuclear leukocytes (PBMC), was isolated using a monoclonal antibody to the protein and radioiodinated by the chloramine-T method. Effective labelling of the protein with little loss of bioactivity was obtained. The labelled protein was found to bind specifically to high affinity receptors present on cells of various cultured lines. The molecular nature of these receptors was examined using the bifunctional cross-linking reagents disuccinimidyl suberate (DSS) and dithiobis succinimidyl propionate (DSP). In SDS-PAGE, conjugates between TNF receptors and the labelled TNF of two sizes (about 75 and 92 kDa) were detected. 125I-labelled TNF was also used to examine variations in TNF receptor concentration, upon treatment of cells by interferon (IFN) or TNF, and any correlation of observed variations with the effect of IFN and TNF on responsiveness of cells to the cytotoxicity of TNF. The decreased vulnerability of cells to the cytotoxic effect of TNF, following treatment of cells by TNF itself, was not correlated with decreased availability of free receptors for the protein. However, an increase in responsiveness of cells to TNF following treatment by IFN was accompanied by some increase in binding of TNF to the receptors.
mesh_terms: D002460:Cell Line; D003429:Cross Reactions; D003601:Cytotoxicity Tests, Immunologic / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005347:Fibroblasts / Q000378:metabolism / Q000648:ultrastructure; D006023:Glycoproteins / Q000096:biosynthesis / Q000302:isolation & purification* / Q000502:physiology; D006367:HeLa Cells / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D006801:Humans; D007372:Interferons / Q000494:pharmacology*; D007457:Iodine Radioisotopes; D008214:Lymphocytes / Q000276:immunology / Q000378:metabolism; D008970:Molecular Weight; D011956:Receptors, Cell Surface / Q000378:metabolism*; D018124:Receptors, Tumor Necrosis Factor; D014409:Tumor Necrosis Factor-alpha
chemical_list: D006023:Glycoproteins; D007457:Iodine Radioisotopes; D011956:Receptors, Cell Surface; D018124:Receptors, Tumor Necrosis Factor; D014409:Tumor Necrosis Factor-alpha; D007372:Interferons