Inhibition of Vero Cell Cytotoxic Activity in Eescherichia coli O157:H7 Lysates by Globotriaosylceramide, Gb3 from Bovine Milk

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  • WATARAI Shinobu
    Laboratory of Veterinary Immunology, Division of Veterinary Science, Graduate School of Agriculture and Biological Sciences, Osaka Prefecture University
  • Tana
    Laboratory of Veterinary Immunology, Division of Veterinary Science, Graduate School of Agriculture and Biological Sciences, Osaka Prefecture University
  • INOUE Kaoru
    Department of Bacteriology, Okayama University Medical School
  • KUSHI Yasunori
    Department of Biochemistry, Faculty of Medicine, Tokyo Medical and Dental University
  • ISOGAI Emiko
    Department of Preventive Dentistry, Health Sciences University of Hokkaido
  • YOKOTA Kenji
    Department of Bacteriology, Okayama University Medical School
  • NAKA Katsumi
    Department of Cell Chemistry, Institute of Cellular and Molecular Biology, Okayama University Medical School
  • OGUMA Keiji
    Department of Bacteriology, Okayama University Medical School
  • KODAMA Hiroshi
    Laboratory of Veterinary Immunology, Division of Veterinary Science, Graduate School of Agriculture and Biological Sciences, Osaka Prefecture University

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  • Inhibition of Vero Cell Cytotoxic Activity in Escherichia coli O157:H7 Lysates by Globotriaosylceramide, Gb3, from Bovine Milk

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In order to clarify the presence and verotoxin (VT) inhibitory activity of globotriaosylceramide (Gb3) in bovine milk, we analyzed neutral glycosphingolipids (GSLs) from bovine milk and investigated the inhibitory effect of bovine milk Gb3 on the cytotoxicity of VT2. Five species of neutral GSLs, designated as N-1, N-2, N-3, N-4, and N-5, were separated on thin-layer chromatography (TLC). N-1, N-2, and N-3 showed the same mobility as glucosylceramide, lactosylceramide, and Gb3 on the TLC plate, respectively. N-4 and N-5 GSLs migrated below globoside on the TLC plate. N-3 GSL having the same TLC mobility as Gb3 from bovine milk was immunologically identified as Gb3 by monoclonal antibody against Gb3, anti-CD77 monoclonal antibody. Furthermore, the effect of bovine milk Gb3 on VT2-induced cytotoxicity was investigated. We found that treatment of VT2 with bovine milk Gb3 can reduce the cytotoxic effect of VT2.

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