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Characterization of a monoclonal antibody reactive with a glycolipid antigen expressed by tumorigenic and certain immortalized, non-tumorigenic rat esophageal epithelial cell lines

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Abstract

A monoclonal antibody (mAb 5G) was produced against a tumorigenic rat esophageal epithelial cell line, designated B2T. Using an enzyme-linked immunosorbent assay, immunofluorescence assay (IFA), thin-layer chromatography (TLC) and immunoperoxidase staining, it was found that mAb 5G reacted specifically with a glycolipid antigen expressed by three tumorigenic rat esophageal epithelial cell lines, and two out of the three nontumorigenic, immortalized rat esophageal epithelial cell lines tested; but did not react with primary cultures of normal rat esophageal epithelial cells or fibroblasts. mAb 5G did not bind to rat respiratory tract carcinoma cell lines, to immortalized rat tracheal epithelial cell lines, or to primary cultures of normal rat tracheal epithelial cells. In addition, mAb 5G did not react with any of the human or mouse cell lines tested. In IFA experiments, mAb 5G stained imprints prepared from in vivo propagated B2T tumor tissues, but did not react with normal rat esophageal, tracheal, lung, liver, and kidney tissues. The antigen was identified by TLC as a neutral glycolipid, consisting of two bands, withR F = 0.45 and 0.41, which migrated in proximity to the ceramide trihexoside standard on TLC plates. Densitometric scanning of the antigen bands indicated that the tumorigenic rat esophageal cell lines possessed 50%–90% more mAb-5G-reactive antigen than the nontumorigenic esophageal cell lines. The results show that mAb 5G reacts specifically with a glycolipid antigen expressed by tumorigenic and certain non-tumorigenic, immortalized rat esophageal epithelial cell lines that might be at the late stages of transformation and early malignancy.

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Author notes

  1. Xingsheng Wan

    Present address: Graduate Center for Toxicology, University of Kentucky, 40 506-0054, Lexington, Kentucky, USA

Authors and Affiliations

  1. Department of Biological Sciences, Bowling Green State University, 43 403, Bowling Green, OH, USA

    Xingsheng Wan & Roudabeh J. Jamasbi

  2. Department of Medical Technology, Bowling Green State University, 43 403, Bowling Green, OH, USA

    Roudabeh J. Jamasbi

  3. Medical College of Ohio, 43 699, Toledo, OH, USA

    Gary D. Stoner

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  1. Xingsheng Wan
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  2. Roudabeh J. Jamasbi
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  3. Gary D. Stoner
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Wan, X., Jamasbi, R.J. & Stoner, G.D. Characterization of a monoclonal antibody reactive with a glycolipid antigen expressed by tumorigenic and certain immortalized, non-tumorigenic rat esophageal epithelial cell lines. Cancer Immunol Immunother 36, 94–100 (1993). https://doi.org/10.1007/BF01754408

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  • DOI: https://doi.org/10.1007/BF01754408

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