Abstract
Detection and localization of HCV in liver tissue are vital for diagnostic purposes and clinical management of HCV-infected patients, as well as for the elucidation of viropathological mechanisms. The fragility of HCV RNA and the low levels of viral expression in infected tissues are a constant limitation in molecular assays for HCV characterization. HCV antigen detection, by immunochemistry, in liver biopsies is an attractive option for precise localization and quantification of viral proteins with direct access to histological patterns. We describe here a study using a novel immunohistochemical method effective on fixed, archived specimens, including liver biopsies and surgical resection samples. The initial protocol uses a biotin-detection system but can also be used in a polymer-detection system. This protocol offers easy, precise, and strong staining resolution with distinct patterns consistent with the liver pathology, irrespective of the viral HCV genotype examined. This approach provides applications for diagnosis as well as for exploratory pathological studies.
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References
Shimizu, Y. K., Feinstone, S. M., Kohara, M., Purcell, R. H., and Yoshikura, H. (1996) Hepatitis C virus: detection of intracellular virus particles by electron microscopy. Hepatology 23, 205–209.
Negro, F. (1998) Detection of hepatitis C virus RNA in liver tissue: an overview. Ital. J. Gastroenterol. Hepatol. 30, 205–210.
Lau, J. Y., Krawczynski, K., Negro, F., Gonzalez Peralta. R. P. (1996) In situ detection of hepatitis C virus—a critical appraisal. J. Hepatol. 24(Suppl. 2), 43–51.
Biagini, P., Benkoel, L., Dodero, F., de Lamballerie, X., Chamlian, V., Nouhou, H., et al. (2001) Hepatitis C virus RNA detection by in situ RT-PCR in formalin-fixed paraffin-embedded liver tissue. Comparison with serum and tissue results. Cell. Mol. Biol. (Noisy-le-Grand), 47 Online Pub OL167-71.
Comar, M., Dal Molin, G., D’Agaro, P., Croce, S. L., Tiribelli, C., and Campello, C. (2006) HBV, HCV, and TTV detection by in situ polymerase chain reaction could reveal occult infection in hepatocellular carcinoma: comparison with blood markers. J. Clin. Pathol. 59, 526–529.
Komminoth, P., Adams, V., Long, A. A., Roth, J., Saremaslani, P., Flury, R., et al. (1994) Evaluation of methods for hepatitis C virus detection in archival liver biopsies. Comparison of histology, immunohistochemistry, in situ hybridization, reverse transcriptase polymerase chain reaction (RT-PCR) and in situ RT-PCR. Pathol. Res. Pract. 190, 1017–1025.
Walker, F. M., Dazza, M. C., Dauge, M. C., Boucher, O., Bedel, C., Henin, D., et al. (1998) Detection and localization by in situ molecular biology techniques and immunohistochemistry of hepatitis C virus in livers of chronically infected patients. J. Histochem. Cytochem. 46, 653–660.
Brody, R. I., Eng, S., Melamed, J., Mizrachi, H., Schneider, R. J., Tobias, H., et al. (1998) Immunohistochemical detection of hepatitis C antigen by monoclonal antibody TORDJI-22 compared with PCR viral detection. Am. J. Clin. Pathol. 110, 32–37.
Blight, K., Lesniewski, R., Labrooy, J., Trowbridge, R., and Gowans, E. (1993) Localisation of hepatitis C virus proteins in infected liver tissue by immunofluorescence. Gastroenterol. Jpn. 28(Suppl 5), 55–58.
Gonzalez-Peralta, R. P., Fang, J. W. S., Davis, G. L., Gish, R., Tsukiyama-Kohara, K., Kohara, M, et al. (1994) Optimization for the detection of hepatitis C virus antigens in the liver. J. Hepatol. 20, 143–147.
Verslype, C., Nevens, F., Sinelli, N., Clarysse, C., Pirenne, J., Depla, E., et al. (2003) Hepatic immunohistochemical staining with a monoclonal antibody against HCV-E2 to evaluate antiviral therapy and reinfection of liver grafts in hepatitis C viral infection. J. Hepatol. 38, 208–214.
Galy, O., Petit, M. A., Benjelloun, S., Chevallier, P., Chevallier, M., Srivatanakul, P., et al. (2007) Efficient hepatitis C antigen immunohistological staining in sections of normal, cirrhotic and tumoral liver using a new monoclonal antibody directed against serum-derived HCV E2 glycoproteins. Cancer Lett. 248, 81–88.
Penin, F., Combet, C., Germanidis, G., Frainais, P. O., Deleage, G., and Pawlotsky, J. M. (2001) Conservation of the conformation and positive charges of hepatitis C virus E2 envelope glycoprotein hypervariable region 1 points to a role in cell attachment. J. Virol. 75, 5703–5710.
Acknowledgements
We thank M. A. Petit for providing the anti-E2 Abs and C. Trépo for his constant interest and support.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Galy, O., Vincent, I., Chevallier, P., Lyandrat, N., Chemin, I. (2009). Immunohistochemical Detection of HCV Proteins in Liver Tissue. In: Tang, H. (eds) Hepatitis C. Methods in Molecular Biology™, vol 510. Humana Press. https://doi.org/10.1007/978-1-59745-394-3_3
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DOI: https://doi.org/10.1007/978-1-59745-394-3_3
Publisher Name: Humana Press
Print ISBN: 978-1-58829-970-3
Online ISBN: 978-1-59745-394-3
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