Antibodies against amino acids 1-15 of tumor necrosis factor block its binding to cell-surface receptor.
Human tumor necrosis factor (hTNF) mediates a variety of biologic activities, which are dependent on the attachment of hTNF to cell-surface receptors. To identify regions of the hTNF protein involved in binding hTNF to its receptor, we prepared five synthetic peptides [hTNF-(1-15), hTNF-(1-31), hTNF-(65-79), hTNF-(98-111), and hTNF-(124-141)] and two hydroxylamine cleavage fragments [hTNF-(1-39) and hTNF-(40-157)] of hTNF. The hTNF-synthetic peptides and hTNF fragments were tested in hTNF receptor binding assays and in two biologic assays: cytolysis of tumor cells and suppression of lipoprotein lipase in adipocytes. Neither the synthetic peptides nor hTNF fragments were active agonists or antagonists in these assays. The synthetic peptides were also conjugated to thyroglobulin, and peptide-specific antisera were raised. All five peptide-thyroglobulin conjugates induced antibody responses to the immunizing peptide and to hTNF. Each antiserum was tested for antagonist activity in hTNF binding assays. Only antisera raised against hTNF-(1-15) or hTNF-(1-31) and antisera against whole hTNF blocked binding. IgGs purified from these three antisera also block hTNF-induced cytolysis and lipoprotein lipase suppression. We conclude that antibodies that recognize the N-terminus of hTNF block the attachment of hTNF to its cellular receptor and inhibit the biologic effects of hTNF.
Socher
S H
SH
Department of Virus and Cell Biology, Merck Sharp & Dohme Research Laboratories, West Point, PA 19486.
Riemen
M W
MW
Martinez
D
D
Friedman
A
A
Tai
J
J
Quintero
J C
JC
Garsky
V
V
Oliff
A
A
D000937:Antigen-Antibody Reactions; D001681:Biological Assay; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D008071:Lipoprotein Lipase / Q000378:metabolism; D009842:Oligopeptides / Q000138:chemical synthesis / Q000276:immunology; D011863:Radioimmunoassay; D011869:Radioligand Assay; D011956:Receptors, Cell Surface / Q000378:metabolism*; D018124:Receptors, Tumor Necrosis Factor; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology*
D009842:Oligopeptides; D011956:Receptors, Cell Surface; D018124:Receptors, Tumor Necrosis Factor; D014409:Tumor Necrosis Factor-alpha; D008071:Lipoprotein Lipase