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A novel monoclonal antibody for detection of galectin-9 in tissue sections : application to human tissues infected by oncogenic viruses.

Abstract : ABSTRACT: BACKGROUND: Galectin-9 is a mammalian lectin which possesses immunosuppressive properties. Excessive production of galectin-9 has been reported in two types of human virus-associated diseases : chronic hepatitis C and nasopharyngeal carcinoma associated to the Epstein-Barr virus. The objective of this study was to produce new monoclonal antibodies targeting galectin-9 in order to improve its detection in clinical samples especially on tissue sections analysed by immunohistochemistry. METHODS: Hybridomas were produced through immunization of mice with the recombinant c-terminal part of galectin-9 (residues 149 to 311 of the S-isoform) and semi-solid fusion of spleen cells with Sp2/0 cells. Monoclonal antibodies were characterized using ELISA, epitope mapping, western blot and immunohistochemistry. RESULTS: We selected seven hybridomas producing antibodies reacting with our recombinant c-terminal galectin-9 in ELISA. Five of them reacted with the epitope "TPAIPPMMYPHPA" (residues 166 to 178 of the S-isoform) and stained all three isoforms of galectin-9 analysed by western blot. One of them, 1G3, demonstrated very good sensitivity and specificity when used for immunohistochemistry. Using 1G3, we could confirm the intense and constant expression of galectin-9 by Epstein-Barr virus positive malignant cells from nasopharyngeal carcinomas. In most samples, specific staining was detected in both cytoplasm and nuclei. Galectin-9 was also detected in liver biopsies from patients infected by the human hepatitis C or B viruses with expression not only in inflammatory leucocytes and Kupffer cells, but also in hepatocytes. In contrast, galectin-9 was virtually absent in non-infected liver specimens. CONCLUSION: The 1G3 monoclonal antibody will be a powerful tool to assess galectin-9 expression and distribution especially in diseases related to oncogenic viruses.
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Submitted on : Wednesday, September 12, 2012 - 9:08:29 PM
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Clément Barjon, Toshiro Niki, Benjamin Vérillaud, Paule Opolon, Pierre Bedossa, et al.. A novel monoclonal antibody for detection of galectin-9 in tissue sections : application to human tissues infected by oncogenic viruses.. Infectious Agents and Cancer, BioMed Central, 2012, 7 (1), pp.16. ⟨10.1186/1750-9378-7-16⟩. ⟨inserm-00731480⟩

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