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Journal of Histochemistry and Cytochemistry, Vol. 49, 759-766, June 2001, Copyright © 2001, The Histochemical Society, Inc.

ARTICLE

Immunohistochemical Detection of Human Gastrointestinal Glutathione Peroxidase in Normal Tissues and Cultured Cells with Novel Mouse Monoclonal Antibodies

Hiroyoshi Komatsua, Isao Okayasub, Hiroyuki Mitomib, Hirotaka Imaic, Yasuhito Nakagawac, and Fumiya Obataa
a Department of Immunology, School of Allied Health Sciences, Kitasato University, Minato, Tokyo, Japan
b Department of Pathology, School of Medicine, Kitasato University, Minato, Tokyo, Japan
c Kitasato University, Sagamihara, Kanagawa, and Laboratory of Hygienic Chemistry, School of Pharmaceutical Sciences, Kitasato University, Minato, Tokyo, Japan

Correspondence to: Hiroyoshi Komatsu, Dept. of Immunology, School of Allied Health Sciences, Kitasato University, 1-15-1 Kitasato, Sagamihara, Kanagawa 228-8555, Japan. E-mail: hkomatsu@ahs.kitasato-u.ac.jp

This is the first report to describe the successful detection of human gastrointestinal glutathione peroxidase in normal tissues by Western blotting and immunohistochemical staining techniques. Four hybridoma clones producing monoclonal antibodies (MAbs) against the human gastrointestinal glutathione peroxidase were established from mice immunized with a gastrointestinal glutathione peroxidase-derived peptide. The MAbs did not crossreact with other members of the glutathione peroxidase family, be it cellular glutathione peroxidase, phospholipid hydroperoxide glutathione peroxidase, or extracellular glutathione peroxidase. Although the MAbs were found to react with a 24-kD protein in a Western blotting assay using gastric carcinoma cell extracts as antigen, they did not react with a B-lymphoblastoid cell extract. Immunohistochemical staining showed gastrointestinal glutathione peroxidase localized in the cytoplasm and in the nucleus of gastric carcinoma cells. Moreover, gastrointestinal glutathione peroxidase was detected in tissue extracts of human stomach, small intestine, large intestine, liver, and gallbladder by Western blotting, and its localization was immunohistochemically confirmed in the mucosal epithelia of the basal area of gastric pits and intestinal crypts.

(J Histochem Cytochem 49:759–766, 2001)

Key Words: gastrointestinal glutathione, peroxidase, monoclonal antibody, immunohistochemical staining, Western blotting, gastric carcinoma cell


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