The in vitro cytotoxicity of native and modified trichloroacetic acid-extracted antigens from Fusobacterium necrophorum for mouse peritoneal macrophages |
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Authors: | Richard D Garman Michael F Lyons Kelton P Hepper George W Teresa |
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Institution: | 1. Department of Microbiology, University of Minnesota, Minneapolis, MN 55455, U.S.A.;3. Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD 20014, U.S.A.;4. Department of Microbiology, University of Saskatchewan, Saskatoon, Canada |
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Abstract: | A Boivin lipopolysaccharide-protein complex (LPS-P), Westphal LPS (W-LPS), Boivin LPS (B-LPS), and a lipid A-associated protein (LAP) were extracted from Fusobacterium necrophorum bov 5 strain and chemically analyzed for protein, total nitrogen, hydrolyzed hexosamine, and nucleic acid content. Dose and time effects on the in vitro cytotoxicity of these preparations for mouse peritoneal macrophages were determined. Macrophage monolayers were exposed to 2.0 ml of 10, 100, 250, 500, or 1000 μg/ml solutions of the individual preparations for 1, 2, 4, or 6 hr at 37°C, and relative toxicities were determined; B-LPS> LPS-P> LAP, W-LPS. The effects of heat, NaOH, and trypsin and pronase on the native antigens were determined. Heat treatment had no effect on the cytotoxicity of B-LPS, but increased the toxicity of LPS-P and LAP for macrophages. Alkaline treatment decreased the cytotoxicity of B-LPS and increased the toxicity of LPS-P and LAP for macrophages. Enzyme treatment had no effect on LPS-P toxicity. The relationship of lipid A and protein content to toxicity for macrophages is discussed. Lipid A appears to be important in the cytotoxicity of B-LPS, and the protein components of LPS-P and LAP may contribute to their toxic activity. The ability of these bacterial cellular antigens to destroy phagocytic cells may facilitate the establishment of necrotic abscesses in susceptible hosts. |
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Keywords: | Fusobacterium necrophorum trichloroacetic acid-extracted antigens cytotoxicity for macrophagesAuthor Keywords: Fusobacterium necrophorum antigènes extraits par l'acide trichloroacétique cytotoxicité sur les macrophages |
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