This study analysed the immune response in the intestinal tract of ducks infected with low-pathogenic avian influenza viruses compared with ducks treated orally with R848, a synthetic Toll-like receptor 7 (TLR7) agonist. Influenza virus infection induced a type I interferon (IFN)-dependent immune response characterized by the expression of Mx transcripts in the ileum at levels that were proportional to viral load. Mx transcripts were detected in differentiated enterocytes from influenza virus-infected ducks. By contrast, in R848-treated ducks, Mx transcripts were detected solely in intraepithelial round cells of haematopoietic origin. An increase was detected in the number of intraepithelial TLR7-positive cells and intraepithelial IFN--producing cells in influenza virus-infected ducks, albeit to a lower level than in R848-treated ducks. IFN- expression was also upregulated in the intestine of influenza virus-infected and R848-treated ducks. Finally, interleukin (IL)-1 and IL-8 transcripts were expressed at high levels in R848-treated ducks but were not increased in influenza virus-infected ducks. These findings suggest that a type I IFN-mediated immune response in enterocytes and the activation of IFN--secreting cells contribute to the control of influenza virus replication in the duck intestine.


Article metrics loading...

Loading full text...

Full text loading...



  1. Adams, S. C., Xing, Z., Li, J. & Cardona, C. J.(2009). Immune-related gene expression in response to H11N9 low pathogenic avian influenza virus infection in chicken and Pekin duck peripheral blood mononuclear cells. Mol Immunol 46, 1744–1749.[CrossRef] [Google Scholar]
  2. Barber, M. R., Aldridge, J. R., Jr, Webster, R. G. & Magor, K. E.(2010). Association of RIG-I with innate immunity of ducks to influenza. Proc Natl Acad Sci U S A 107, 5913–5918.[CrossRef] [Google Scholar]
  3. Bazzigher, L., Schwarz, A. & Staeheli, P.(1993). No enhanced influenza virus resistance of murine and avian cells expressing cloned duck Mx protein. Virology 195, 100–112.[CrossRef] [Google Scholar]
  4. Bernier, B., Bar, I., Pieau, C., Lambert De Rouvroit, C. & Goffinet, A. M.(1999). Reelin mRNA expression during embryonic brain development in the turtle Emys orbicularis. J Comp Neurol 413, 463–479.[CrossRef] [Google Scholar]
  5. Brown, C.(1998).In situ hybridization with riboprobes: an overview for veterinary pathologists. Vet Pathol 35, 159–167.[CrossRef] [Google Scholar]
  6. De Marco, M. A., Foni, E., Campitelli, L., Delogu, M., Raffini, E., Chiapponi, C., Barigazzi, G., Cordioli, P., Di Trani, L. & Donatelli, I.(2005). Influenza virus circulation in wild aquatic birds in Italy during H5N2 and H7N1 poultry epidemic periods (1998–2000). Avian Pathol 34, 480–485.[CrossRef] [Google Scholar]
  7. Hale, B. G., Randall, R. E., Ortín, J. & Jackson, D.(2008). The multifunctional NS1 protein of influenza A viruses. J Gen Virol 89, 2359–2376.[CrossRef] [Google Scholar]
  8. Heil, F., Hemmi, H., Hochrein, H., Ampenberger, F., Kirschning, C., Akira, S., Lipford, G., Wagner, H. & Bauer, S.(2004). Species-specific recognition of single-stranded RNA via Toll-like receptor 7 and 8. Science 303, 1526–1529.[CrossRef] [Google Scholar]
  9. Hemmi, H., Kaisho, T., Takeuchi, O., Sato, S., Sanjo, H., Hoshino, K., Horiuchi, T., Tomizawa, H., Takeda, K. & Akira, S.(2002). Small anti-viral compounds activate immune cells via the TLR7 MyD88-dependent signaling pathway. Nat Immunol 3, 196–200.[CrossRef] [Google Scholar]
  10. Holzinger, D., Jorns, C., Stertz, S., Boisson-Dupuis, S., Thimme, R., Weidmann, M., Casanova, J. L., Haller, O. & Kochs, G.(2007). Induction of MxA gene expression by influenza A virus requires type I or type III interferon signaling. J Virol 81, 7776–7785.[CrossRef] [Google Scholar]
  11. Iqbal, M., Philbin, V. J. & Smith, A. L.(2005). Expression patterns of chicken Toll-like receptor mRNA in tissues, immune cell subsets and cell lines. Vet Immunol Immunopathol 104, 117–127.[CrossRef] [Google Scholar]
  12. Ito, T., Amakawa, R., Kaisho, T., Hemmi, H., Tajima, K., Uehira, K., Ozaki, Y., Tomizawa, H., Akira, S. & Fukuhara, S.(2002). Interferon-α and interleukin-12 are induced differentially by Toll-like receptor 7 ligands in human blood dendritic cell subsets. J Exp Med 195, 1507–1512.[CrossRef] [Google Scholar]
  13. Jurk, M., Heil, F., Vollmer, J., Schetter, C., Krieg, A. M., Wagner, H., Lipford, G. & Bauer, S.(2002). Human TLR7 or TLR8 independently confer responsiveness to the antiviral compound R-848. Nat Immunol 3, 499.[CrossRef] [Google Scholar]
  14. Karlsson, A., Jägervall, K., Utkovic, H., Karlsson, L., Rehnström, E., Fredin, M. F., Gillberg, P. G., Jansson, L., Michaëlsson, E. & Melgar, S.(2008). Intra-colonic administration of the TLR7 agonist R-848 induces an acute local and systemic inflammation in mice. Biochem Biophys Res Commun 367, 242–248.[CrossRef] [Google Scholar]
  15. Karpala, A. J., Morris, K. R., Broadway, M. M., McWaters, P. G., O'Neil, T. E., Goossens, K. E., Lowenthal, J. W. & Bean, A. G.(2008). Molecular cloning, expression, and characterization of chicken IFN-λ. J Interferon Cytokine Res 28, 341–350.[CrossRef] [Google Scholar]
  16. Kida, H., Yanagawa, R. & Matsuoka, Y.(1980). Duck influenza lacking evidence of disease signs and immune response. Infect Immun 30, 547–553. [Google Scholar]
  17. Kim, J. K., Seiler, P., Forrest, H. L., Khalenkov, A. M., Franks, J., Kumar, M., Karesh, W. B., Gilbert, M., Sodnomdarjaa, R. & other authors(2008). Pathogenicity and vaccine efficacy of different clades of Asian H5N1 avian influenza A viruses in domestic ducks. J Virol 82, 11374–11382.[CrossRef] [Google Scholar]
  18. Kochs, G., García-Sastre, A. & Martínez-Sobrido, L.(2007). Multiple anti-interferon actions of the influenza A virus NS1 protein. J Virol 81, 7011–7021.[CrossRef] [Google Scholar]
  19. Kumagai, Y., Takeuchi, O., Kato, H., Kumar, H., Matsui, K., Morii, E., Aozasa, K., Kawai, T. & Akira, S.(2007). Alveolar macrophages are the primary interferon-alpha producer in pulmonary infection with RNA viruses. Immunity 27, 240–252.[CrossRef] [Google Scholar]
  20. Löndt, B. Z., Núñez, A., Banks, J., Alexander, D. J., Russell, C., Richard-Löndt, A. C. & Brown, I. H.(2010). The effect of age on the pathogenesis of a highly pathogenic avian influenza (HPAI) H5N1 virus in Pekin ducks (Anas platyrhynchos) infected experimentally. Influenza Other Respir Viruses 4, 17–25.[CrossRef] [Google Scholar]
  21. MacDonald, M. R., Xia, J., Smith, A. L. & Magor, K. E.(2008). The duck Toll like receptor 7: genomic organization, expression and function. Mol Immunol 45, 2055–2061.[CrossRef] [Google Scholar]
  22. Mordstein, M., Neugebauer, E., Ditt, V., Jessen, B., Rieger, T., Falcone, V., Sorgeloos, F., Ehl, S., Mayer, D. & other authors(2010). Lambda interferon renders epithelial cells of the respiratory and gastrointestinal tracts resistant to viral infections. J Virol 84, 5670–5677.[CrossRef] [Google Scholar]
  23. Munster, V. J., Baas, C., Lexmond, P., Waldenström, J., Wallensten, A., Fransson, T., Rimmelzwaan, G. F., Beyer, W. E., Schutten, M. & other authors(2007). Spatial, temporal, and species variation in prevalence of influenza A viruses in wild migratory birds. PLoS Pathog 3, e61.[CrossRef] [Google Scholar]
  24. Olsen, B., Munster, V. J., Wallensten, A., Waldenström, J., Osterhaus, A. D. & Fouchier, R. A.(2006). Global patterns of influenza A virus in wild birds. Science 312, 384–388.[CrossRef] [Google Scholar]
  25. Pantin-Jackwood, M. J., Suarez, D. L., Spackman, E. & Swayne, D. E.(2007). Age at infection affects the pathogenicity of Asian highly pathogenic avian influenza H5N1 viruses in ducks. Virus Res 130, 151–161.[CrossRef] [Google Scholar]
  26. Peirson, S. N., Butler, J. N. & Foster, R. G.(2003). Experimental validation of novel and conventional approaches to quantitative real-time PCR data analysis. Nucleic Acids Res 31, e73.[CrossRef] [Google Scholar]
  27. Perkins, L. E. & Swayne, D. E.(2003). Comparative susceptibility of selected avian and mammalian species to a Hong Kong-origin H5N1 high-pathogenicity avian influenza virus. Avian Dis 47 (Suppl.), 956–967.[CrossRef] [Google Scholar]
  28. Pockros, P. J., Guyader, D., Patton, H., Tong, M. J., Wright, T., McHutchison, J. G. & Meng, T. C.(2007). Oral resiquimod in chronic HCV infection: safety and efficacy in 2 placebo-controlled, double-blind phase IIa studies. J Hepatol 47, 174–182.[CrossRef] [Google Scholar]
  29. Reemers, S. S., van Leenen, D., Groot Koerkamp, M. J., van Haarlem, D., van de Haar, P., van Eden, W. & Vervelde, L.(2010). Early host responses to avian influenza A virus are prolonged and enhanced at transcriptional level depending on maturation of the immune system. Mol Immunol 47, 1675–1685.[CrossRef] [Google Scholar]
  30. Schultz, U. & Chisari, F. V.(1999). Recombinant duck interferon gamma inhibits duck hepatitis B virus replication in primary hepatocytes. J Virol 73, 3162–3168. [Google Scholar]
  31. Schultz, U., Köck, J., Schlicht, H. J. & Staeheli, P.(1995). Recombinant duck interferon: a new reagent for studying the mode of interferon action against hepatitis B virus. Virology 212, 641–649.[CrossRef] [Google Scholar]
  32. Smit, J. J., Rudd, B. D. & Lukacs, N. W.(2006). Plasmacytoid dendritic cells inhibit pulmonary immunopathology and promote clearance of respiratory syncytial virus. J Exp Med 203, 1153–1159.[CrossRef] [Google Scholar]
  33. Sommereyns, C., Paul, S., Staeheli, P. & Michiels, T.(2008). IFN-lambda (IFN-λ) is expressed in a tissue-dependent fashion and primarily acts on epithelial cells in vivo. PLoS Pathog 4, e1000017.[CrossRef] [Google Scholar]
  34. Soubies, S. M., Volmer, C., Croville, G., Loupias, J., Peralta, B., Costes, P., Lacroux, C., Guérin, J.-L. & Volmer, R.(2010a). Species-specific contribution of the four C-terminal amino acids of influenza A virus NS1 protein to virulence. J Virol 84, 6733–6747.[CrossRef] [Google Scholar]
  35. Soubies, S. M., Volmer, C., Guérin, J. L. & Volmer, R.(2010b). Truncation of the NS1 protein converts a low pathogenic avian influenza virus into a strong interferon inducer in duck cells. Avian Dis 54 (Suppl.), 527–531.[CrossRef] [Google Scholar]
  36. Sturm-Ramirez, K. M., Hulse-Post, D. J., Govorkova, E. A., Humberd, J., Seiler, P., Puthavathana, P., Buranathai, C., Nguyen, T. D., Chaisingh, A. & other authors(2005). Are ducks contributing to the endemicity of highly pathogenic H5N1 influenza virus in Asia? J Virol 79, 11269–11279.[CrossRef] [Google Scholar]
  37. Taubenberger, J. K. & Kash, J. C.(2010). Influenza virus evolution, host adaptation, and pandemic formation. Cell Host Microbe 7, 440–451.[CrossRef] [Google Scholar]
  38. van Pesch, V., Lanaya, H., Renauld, J. C. & Michiels, T.(2004). Characterization of the murine alpha interferon gene family. J Virol 78, 8219–8228.[CrossRef] [Google Scholar]
  39. Webster, R. G., Yakhno, M., Hinshaw, V. S., Bean, W. J. & Murti, K. G.(1978). Intestinal influenza: replication and characterization of influenza viruses in ducks. Virology 84, 268–278.[CrossRef] [Google Scholar]
  40. Wood, G. W., Parsons, G. & Alexander, D. J.(1995). Replication of influenza A viruses of high and low pathogenicity for chickens at different sites in chickens and ducks following intranasal inoculation. Avian Pathol 24, 545–551.[CrossRef] [Google Scholar]
  41. Wu, Y. F., Liu, H. J., Chiou, S. H. & Lee, L. H.(2007). Sequence and phylogenetic analysis of interleukin (IL)-1β-encoding genes of five avian species and structural and functional homology among these IL-1β proteins. Vet Immunol Immunopathol 116, 37–46.[CrossRef] [Google Scholar]
  42. Wu, Y. F., Shien, J. H., Yin, H. H., Chiow, S. H. & Lee, L. H.(2008). Structural and functional homology among chicken, duck, goose, turkey and pigeon interleukin-8 proteins. Vet Immunol Immunopathol 125, 205–215.[CrossRef] [Google Scholar]
  43. Yrlid, U., Milling, S. W., Miller, J. L., Cartland, S., Jenkins, C. D. & MacPherson, G. G.(2006). Regulation of intestinal dendritic cell migration and activation by plasmacytoid dendritic cells, TNF-α and type 1 IFNs after feeding a TLR7/8 ligand. J Immunol 176, 5205–5212.[CrossRef] [Google Scholar]

Data & Media loading...


vol. , part 3, pp. 534 - 543

Primer sequences used in this study [PDF](80 KB)


Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error