This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Castrucci, M R Articles by Kawaoka, Y Search for Related Content PubMed PubMed Citation Articles by Castrucci, M R Articles by Kawaoka, Y

 Previous Article  |  Next Article 

J Virol. 1992 August; 66(8): 4647-4653

Attenuation of influenza A virus by insertion of a foreign epitope into the neuraminidase.

Department of Virology and Molecular Biology, St. Jude Children's Research Hospital, Memphis, Tennessee 38101-0318.

ABSTRACT

As the initial step in generating a live attenuated influenza A vaccine, we attempted to substitute an unrelated amino acid sequence (FLAG) for a portion of the neuraminidase (NA) molecule in influenza virus A/WSN/33 (H1N1), using a recently developed technique (reverse genetics [W. Luytjes, M. Krystal, M. Enami, J. D. Parvin, and P. Palese, Cell 59:1107-1113, 1989]). This technique allowed us to rescue the NA molecules containing the FLAG sequence (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys) at the bottom portion of the boxlike head of the molecule immediately above the stalk region (amino acid residues 63 to 70 [WSN NA numbering]). An anti-FLAG monoclonal antibody immunoprecipitated the NA molecules with the FLAG sequence, demonstrating that the foreign epitope was exposed on the virion surface. The dose of FLAG-containing transfectant virus required to kill 50% of mice was 100-fold higher than the required dose of parent virus. The FLAG sequence was stably maintained in the NA molecule during passage of the virus in tissue culture and in mice. These findings demonstrate that live influenza A vaccine strains with stable attenuating mutations in the coding region of the viral genes can be generated by reverse genetics.

This article has been cited by other articles:

• Lin, H., Goodenough, U. W. (2007). Gametogenesis in the Chlamydomonas reinhardtii minus Mating Type Is Controlled by Two Genes, MID and MTD1. Genetics 176: 913-925 [Abstract] [Full Text]  
• Taylor, A. P., Goldenberg, D. M. (2007). Role of placenta growth factor in malignancy and evidence that an antagonistic PlGF/Flt-1 peptide inhibits the growth and metastasis of human breast cancer xenografts. Molecular Cancer Therapeutics 6: 524-531 [Abstract] [Full Text]  
• Jenkins, M. R., Webby, R., Doherty, P. C., Turner, S. J. (2006). Addition of a Prominent Epitope Affects Influenza A Virus-Specific CD8+ T Cell Immunodominance Hierarchies When Antigen Is Limiting.. J. Immunol. 177: 2917-2925 [Abstract] [Full Text]  
• Chen, X.-J., Seth, S., Yue, G., Kamat, P., Compans, R. W., Guidot, D., Brown, L. A., Eaton, D. C., Jain, L. (2004). Influenza virus inhibits ENaC and lung fluid clearance. Am. J. Physiol. Lung Cell. Mol. Physiol. 287: L366-L373 [Abstract] [Full Text]  
• Garulli, B., Kawaoka, Y., Castrucci, M. R. (2004). Mucosal and Systemic Immune Responses to a Human Immunodeficiency Virus Type 1 Epitope Induced upon Vaginal Infection with a Recombinant Influenza A Virus. J. Virol. 78: 1020-1025 [Abstract] [Full Text]  
• Swigut, T., Greenberg, M., Skowronski, J. (2003). Cooperative Interactions of Simian Immunodeficiency Virus Nef, AP-2, and CD3-{zeta} Mediate the Selective Induction of T-Cell Receptor-CD3 Endocytosis. J. Virol. 77: 8116-8126 [Abstract] [Full Text]  
• Pathak, M. K., Dhawan, D., Lindner, D. J., Borden, E. C., Farver, C., Yi, T. (2002). Pentamidine Is an Inhibitor of PRL Phosphatases with Anticancer Activity. Molecular Cancer Therapeutics 1: 1255-1264 [Abstract] [Full Text]  
• Neumann, G., Whitt, M. A., Kawaoka, Y. (2002). A decade after the generation of a negative-sense RNA virus from cloned cDNA - what have we learned?. J. Gen. Virol. 83: 2635-2662 [Abstract] [Full Text]  
• Kunzelmann, K., Beesley, A. H., King, N. J., Karupiah, G., Young, J. A., Cook, D. I. (2000). Influenza virus inhibits amiloride-sensitive Na+ channels in respiratory epithelia. Proc. Natl. Acad. Sci. USA 10.1073/pnas.160041997v1 [Abstract] [Full Text]  
• Mitnaul, L. J., Matrosovich, M. N., Castrucci, M. R., Tuzikov, A. B., Bovin, N. V., Kobasa, D., Kawaoka, Y. (2000). Balanced Hemagglutinin and Neuraminidase Activities Are Critical for Efficient Replication of Influenza A Virus. J. Virol. 74: 6015-6020 [Abstract] [Full Text]  
• Solorzano, A., Zheng, H., Fodor, E., Brownlee, G. G., Palese, P., García-Sastre, A. (2000). Reduced levels of neuraminidase of influenza A viruses correlate with attenuated phenotypes in mice. J. Gen. Virol. 81: 737-742 [Abstract] [Full Text]  
• Basler, C. F., Garcia-Sastre, A., Palese, P. (1999). Mutation of Neuraminidase Cysteine Residues Yields Temperature-Sensitive Influenza Viruses. J. Virol. 73: 8095-8103 [Abstract] [Full Text]  
• Neumann, G., Watanabe, T., Ito, H., Watanabe, S., Goto, H., Gao, P., Hughes, M., Perez, D. R., Donis, R., Hoffmann, E., Hobom, G., Kawaoka, Y. (1999). Generation of influenza A viruses entirely from cloned cDNAs. Proc. Natl. Acad. Sci. USA 96: 9345-9350 [Abstract] [Full Text]  
• Roberts, A., Kretzschmar, E., Perkins, A. S., Forman, J., Price, R., Buonocore, L., Kawaoka, Y., Rose, J. K. (1998). Vaccination with a Recombinant Vesicular Stomatitis Virus Expressing an Influenza Virus Hemagglutinin Provides Complete Protection from Influenza Virus Challenge. J. Virol. 72: 4704-4711 [Abstract] [Full Text]  
• Kunzelmann, K., Beesley, A. H., King, N. J., Karupiah, G., Young, J. A., Cook, D. I. (2000). From the Cover: Influenza virus inhibits amiloride-sensitive Na+ channels in respiratory epithelia. Proc. Natl. Acad. Sci. USA 97: 10282-10287 [Abstract] [Full Text]