Induction of tumor necrosis factor and macrophage-mediated cytotoxicity by horseradish peroxidase and other glycosylated proteins: the role of enzymatic activity and LPS.
Recent studies by these investigators have shown that horseradish peroxidase (HRP) can cause murine thioglycollate-induced peritoneal macrophages (M phi) to produce both tumor necrosis factor (TNF) and enhance macrophage-mediated cytotoxicity (MMC) to 3T12 target cells. The present study identifies the roles of both enzymatic activity and contaminating lipopolysaccharides (LPS) (less than or equal to 1 ng) on these activities. The addition of 100 ng/ml of polymyxin B (PB) to enzymatically active HRP significantly reduced TNF production but did not affect MMC. Enzymatically inactive HRP (DHRP) was more effective than HRP in both TNF production and MMC but was not affected by PB. The inability of PB to modify DHRP-induced TNF suggests that LPS was not required. The induction of TNF and MMC in the absence of LPS was also corroborated by similar studies using M phi from endotoxin-resistant C3H/HeJ mice. Glycosylated proteins such as HRP, DHRP, and mannosylated bovine serum albumin (M-BSA) are known to bind to mannose receptors (mannosyl-fucosyl receptor [MFR]) on the surface of M phi. In the present studies, M-BSA behaved similarly to DHRP in that it induced both TNF secretion and MMC. These results suggest that binding to the MFR may be sufficient to induce TNF secretion and MMC. In addition, the data suggest that neither enzymatic activity nor LPS was required for DHRP-induced TNF.
Department of Biological Sciences, Texas Tech University, Lubbock 79409.
D L
DL
Lefkowitz
K
K
Mills
A
A
Castro
S S
SS
Lefkowitz
D000818:Animals; D003602:Cytotoxicity, Immunologic / Q000187:drug effects; D006023:Glycoproteins / Q000494:pharmacology*; D006735:Horseradish Peroxidase / Q000378:metabolism / Q000494:pharmacology*; D007111:Immunity, Cellular / Q000187:drug effects; D008070:Lipopolysaccharides; D008264:Macrophages / Q000276:immunology* / Q000378:metabolism; D008351:Mannans / Q000494:pharmacology; D051379:Mice; D008815:Mice, Inbred Strains; D010529:Peritoneal Cavity / Q000166:cytology; D011112:Polymyxin B / Q000494:pharmacology; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis*
D006023:Glycoproteins; D008070:Lipopolysaccharides; D008351:Mannans; D014409:Tumor Necrosis Factor-alpha; D012710:Serum Albumin, Bovine; D006735:Horseradish Peroxidase; D011112:Polymyxin B