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Open Access Research article

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

Clément Barjon12, Toshiro Niki3, Benjamin Vérillaud1, Paule Opolon1, Pierre Bedossa4, Mitsuomi Hirashima3, Stéphanie Blanchin5, Michel Wassef6, Hugo R Rosen7, Anne-Sophie Jimenez1, Ming Wei2 and Pierre Busson1*

Author Affiliations

1 University Paris-Sud 11, CNRS-UMR 8126, Institut de Cancérologie Gustave Roussy, 114 rue Edouard Vaillant, 94805, Villejuif cedex, France

2 Cellvax, Ecole Nationale Vétérinaire d’Alfort, 7 avenue du Général de Gaulle, 94704, Maisons-Alfort cedex, France

3 Department of Immunology and Immunopathology, Faculty of Medicine, Kagawa University, Kagawa, 761-0793, Japan

4 Département de Pathologie, INSERM U773, Hôpital Beaujon, Université Paris-Diderot, 92110, Clichy, France

5 Px’ Therapeutics, 38040, Grenoble cedex 9, France

6 Département de Pathologie, Hôpital Lariboisière, Université Paris-Diderot, 75475, Paris cedex 10, France

7 Department of Medicine, Division of Gastroenterology & Hepatology, University of Colorado School of Medicine, Aurora, CO, USA

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Infectious Agents and Cancer 2012, 7:16  doi:10.1186/1750-9378-7-16

Published: 17 July 2012

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-terminus part of galectin-9 (residues 191 to 355 of the long 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-terminus galectin-9 in ELISA. Five of them reacted with the epitope “TPAIPPMMYPHPA” (common to all isoforms, residues 210 to 222 of the long 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.