title: Cytotoxic activity and production of toxic nitrogen oxides by macrophages treated with IFN-gamma and monoclonal antibodies against the 73-kDa lipopolysaccharide receptor.
abstract: The hamster IgM mAb 5D3 is specific for an 73-kDa LPS receptor on murine leukocytes. This mAb inhibits binding of radiolabeled LPS to splenocytes and acts as an agonist for induction of LPS-mediated changes in macrophage function. Resident peritoneal macrophages treated with IFN-gamma and mAb 5D3 developed potent cytotoxic activity against tumor cells. Cells treated with IFN-gamma or mAb 5D3 alone were inactive. Macrophage cytotoxic activity induced by IFN-gamma and mAb 5D3 was inhibited by NGMMLA and coincident with high levels of NO2-released into culture fluids. These data show that mAb 5D3 serves as an effective trigger signal for induction of cytotoxic activity with IFN-gamma-primed macrophages. Indeed, mAb 5D3 exactly mimicked the effects of LPS in these same systems. Unlike LPS, effects of mAb 5D3 on induction of macrophage cytotoxic activity and production of nitrogen oxides was abrogated after boiling, and not affected by addition of polymyxin B. The effects of LPS and mAb 5D3 as a trigger signal for IFN-gamma-primed macrophages were associated with production of TNF activity in culture fluids and inhibited by mAb against rTNF-alpha. Expression of class II MHC on macrophages induced by IFN-gamma treatment was suppressed by both LPS and mAb 5D3. These suppressive effects of LPS and mAb 5D3 were not affected by NGMMLA or mAb against rTNF-alpha. Finally, macrophages treated with LPS or mAb 5D3 before exposure to IFN-gamma and LPS or mAb 5D3 did not develop cytotoxic activity or high levels of NO2- in the culture fluids. These same cells developed both effector activities after addition of rTNF-alpha. These results in toto identify the 73-kDa protein as a receptor that mediates LPS-induced changes in macrophage effector function. The mAb 5D3 serves as a specific and defined reagent agonist for analysis of LPS receptor-linked change.
authors.affiliation: Department of Cellular Immunology, Walter Reed Army Institute of Research, Washington, DC 20307-5100.
authors.forename: S J
authors.initials: SJ
authors.lastname: Green
authors.forename: T Y
authors.initials: TY
authors.lastname: Chen
authors.forename: R M
authors.initials: RM
authors.lastname: Crawford
authors.forename: C A
authors.initials: CA
authors.lastname: Nacy
authors.forename: D C
authors.initials: DC
authors.lastname: Morrison
authors.forename: M S
authors.initials: MS
authors.lastname: Meltzer
mesh_terms: D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives / Q000494:pharmacology; D003602:Cytotoxicity, Immunologic; D000949:Histocompatibility Antigens Class II / Q000378:metabolism; D006358:Hot Temperature; D007111:Immunity, Cellular; D007371:Interferon-gamma / Q000494:pharmacology*; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides / Q000008:administration & dosage; D008264:Macrophages / Q000276:immunology*; D051379:Mice; D008809:Mice, Inbred C3H; D009589:Nitrogen Oxides / Q000633:toxicity*; D011112:Polymyxin B / Q000494:pharmacology; D011971:Receptors, Immunologic / Q000502:physiology*; D011994:Recombinant Proteins; D019323:omega-N-Methylarginine
chemical_list: D000949:Histocompatibility Antigens Class II; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D009589:Nitrogen Oxides; D011971:Receptors, Immunologic; D011994:Recombinant Proteins; D019323:omega-N-Methylarginine; D007371:Interferon-gamma; D001120:Arginine; D011112:Polymyxin B