Abstract
Immunohistochemical methods are commonly used for studying the pathogenesis of influenza A virus by allowing the identification of sites of replication of the virus in infected tissues and the correlation with the histopathological changes observed. In this chapter, the materials and procedures for performing immunohistochemical detection of influenza virus antigens in tissues using a biotin–streptavidin detection method are provided. The technique involves the following steps: heat-induced antigen retrieval, binding of a primary antibody to the virus antigen, antibody–antigen complex binding by a biotinylated secondary antibody, and binding of an enzyme–streptavidin conjugate. The enzyme is then visualized by application of the substrate chromogen solution to produce a colorimetric reaction with a product that can be visualized. Visualization of influenza virus antigen in tissues is based on chromogen deposition in the nucleus and cytoplasm of infected cells.
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Ellis TM, Bousfield RB, Bissett LA, Dyrting KC, Luk GS, Tsim ST, Sturm-Ramirez K, Webster RG, Guan Y, Malik Peiris JS (2004) Investigation of outbreaks of highly pathogenic H5N1 avian influenza in waterfowl and wild birds in Hong Kong in late 2002. Avian Pathol 33:492–505
Perkins LE, Swayne DE (2001) Pathobiology of A/chicken/Hong Kong/220/97 (H5N1) avian influenza virus in seven gallinaceous species. Vet Pathol 38:149–164
Perkins LE, Swayne DE (2003) Comparative susceptibility of selected avian and mammalian species to a Hong Kong-origin H5N1 high-pathogenicity avian influenza virus. Avian Dis 47:956–967
Swayne DE, Slemons RD (1995) Comparative pathology of intravenously inoculated wild duck- and turkey-origin type A influenza viruses in chickens. Avian Dis 39:74–84
Nakatani H, Nakamura K, Yamamoto Y, Yamada M, Yamamoto Y (2005) Epidemiology, pathology, and immunohistochemistry of layer hens naturally affected with H5N1 highly pathogenic avian influenza in Japan. Avian Dis 49:436–441
Hooper PT, Russell GW, Selleck PW, Stanislawek WL (1995) Observations on the relationship in chickens between the virulence of some avian influenza viruses and their pathogenicity for various organs. Avian Dis 39:458–464
Kishida N, Sakoda Y, Isoda N, Matsuda K, Eto M, Sunaga Y, Umemura T, Kida H (2005) Pathogenicity of H5 influenza viruses for ducks. Arch Virol 150:1383–1392
Swayne DE (1997) Pathobiology of H5N2 Mexican avian influenza virus infections of chickens. Vet Pathol 34:557–567
Pantin-Jackwood MJ, Swayne DE (2007) Pathobiology of Asian highly pathogenic H5N1 avian influenza virus infection in ducks. Avian Dis 51:250–259.2007
Mo IP, Brugh M, Fletcher OJ, Rowland GN, Swayne DE (1997) Comparative pathology of chickens experimentally inoculated with avian influenza viruses of low and high pathogenicity. Avian Dis 41(1):125–136
Swayne DE, Halvorson DA (2003) Influenza. In: Saif YM, Barnes HJ, Glisson JR, Fadly AM, McDougald LR, Swayne DE (eds) Diseases of poultry, 11th edn. Blackwell, Ames, IA, pp 135–160
Swayne DE, Suarez DL (2000) Highly pathogenic avian influenza. Rev Sci Tech 19(2):463–482
Antarasena C, Sirimujalin R, Prommuang P, Blacksell SD, Promkuntod N, Prommuang P (2006) Tissue tropism of a Thailand strain of high-pathogenicity avian influenza virus (H5N1) in tissues of naturally infected native chickens (Gallus gallus), Japanese quail (Coturnix coturnix japonica) and ducks (Anas spp.). Avian Pathol 35:250–253
Shinya K, Hamm S, Hatta M, Ito H, Ito T, Kawaoka Y (2004) PB2 amino acid at position 627 affects replicative efficiency, but not cell tropism, of Hong Kong H5N1 influenza A viruses in mice. Virology 320:258–266
Ramos-Vara JA (2005) Technical aspects of Immunohistochemistry. Review article. Vet Pathol 42:405–426
Krenacs LTK, Raffeld M (1999) Antigen retrieval for immunohistochemical reactions in routinely processed paraffin sections. In: Lavois LC (ed) Methods in molecular biology: immunocytochemical methods and protocols. Humana Press INC, Totowa, NJ, pp 85–93
Bratthauer GL (1999) The avidin-biotin complex (ABC) method and other avidin-biotin binding methods. In: Javois LC (ed) Methods in molecular biology: immunocytochemical methods and protocols. Humana Press INC, Totowa, NJ, pp 203–214
Acknowledgments
I gratefully acknowledge the assistance of Diane Smith and the personnel from the Food Safety and Inspection Service histopathology laboratory (USDA, Athens, GA) in the development and adaptation of this method.
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Pantin-Jackwood, M.J. (2020). Immunohistochemical Staining of Influenza Virus in Tissues. In: Spackman, E. (eds) Animal Influenza Virus. Methods in Molecular Biology, vol 2123. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0346-8_3
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DOI: https://doi.org/10.1007/978-1-0716-0346-8_3
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