Abstract
We have used a neuraminidase-deficient influenza virus, NWS-Mvi, which was selected by supplying bacterial neuraminidase in the medium (C. Liu and G. M. Air, Virology 194:403-407, 1993), to define the role of neuraminidase in influenza virus replication. Electron microscopy showed that virions of the NWS-Mvi mutant assembled normally and formed large aggregates associated with cell surfaces. The NWS-Mvi virus grown in the absence of neuraminidase was able to carry out a second round of replication in MDCK cells without added neuraminidase, indicating that the virus particles contained in these aggregates were infectious. Aggregates of virus were also found in cytoplasmic vacuoles. When virus-infected cells were incubated in the presence of ferritin, such aggregates were found to be labeled with ferritin, indicating that they are derived from uptake at the cell surface. When the neuraminidase-deficient virus was administered intranasally to C57BL/6 mice, low titers of virus were recovered from the lungs and major histocompatibility complex class I-restricted cytotoxic T cells were generated: evidence that cells were infected in vivo. In C57BL/6 nu/nu mice, the low level of virus persisted for at least 28 days but never increased. These results suggest that neuraminidase is not required for influenza virus entry, replication, or assembly in cell culture or in mice.
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- Air G. M., Laver W. G. The neuraminidase of influenza virus. Proteins. 1989;6(4):341–356. doi: 10.1002/prot.340060402. [DOI] [PubMed] [Google Scholar]
- Allan W., Tabi Z., Cleary A., Doherty P. C. Cellular events in the lymph node and lung of mice with influenza. Consequences of depleting CD4+ T cells. J Immunol. 1990 May 15;144(10):3980–3986. [PubMed] [Google Scholar]
- Compans R. W., Dimmock N. J., Meier-Ewert H. Effect of antibody to neuraminidase on the maturation and hemagglutinating activity of an influenza A2 virus. J Virol. 1969 Oct;4(4):528–534. doi: 10.1128/jvi.4.4.528-534.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Doherty P. C., Allan W., Eichelberger M., Carding S. R. Roles of alpha beta and gamma delta T cell subsets in viral immunity. Annu Rev Immunol. 1992;10:123–151. doi: 10.1146/annurev.iy.10.040192.001011. [DOI] [PubMed] [Google Scholar]
- Griffin J. A., Basak S., Compans R. W. Effects of hexose starvation and the role of sialic acid in influenza virus release. Virology. 1983 Mar;125(2):324–334. doi: 10.1016/0042-6822(83)90205-2. [DOI] [PubMed] [Google Scholar]
- Huang R. T., Dietsch E., Rott R. Further studies on the role of neuraminidase and the mechanism of low pH dependence in influenza virus-induced membrane fusion. J Gen Virol. 1985 Feb;66(Pt 2):295–301. doi: 10.1099/0022-1317-66-2-295. [DOI] [PubMed] [Google Scholar]
- Huang R. T., Rott R., Wahn K., Klenk H. D., Kohama T. The function of the neuraminidase in membrane fusion induced by myxoviruses. Virology. 1980 Dec;107(2):313–319. doi: 10.1016/0042-6822(80)90299-8. [DOI] [PubMed] [Google Scholar]
- Kilbourne E. D. Future influenza vaccines and the use of genetic recombinants. Bull World Health Organ. 1969;41(3):643–645. [PMC free article] [PubMed] [Google Scholar]
- Kilbourne E. D., Laver W. G., Schulman J. L., Webster R. G. Antiviral activity of antiserum specific for an influenza virus neuraminidase. J Virol. 1968 Apr;2(4):281–288. doi: 10.1128/jvi.2.4.281-288.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lamb R. A., Choppin P. W. The gene structure and replication of influenza virus. Annu Rev Biochem. 1983;52:467–506. doi: 10.1146/annurev.bi.52.070183.002343. [DOI] [PubMed] [Google Scholar]
- Li S., Schulman J., Itamura S., Palese P. Glycosylation of neuraminidase determines the neurovirulence of influenza A/WSN/33 virus. J Virol. 1993 Nov;67(11):6667–6673. doi: 10.1128/jvi.67.11.6667-6673.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Liu C., Air G. M. Selection and characterization of a neuraminidase-minus mutant of influenza virus and its rescue by cloned neuraminidase genes. Virology. 1993 May;194(1):403–407. doi: 10.1006/viro.1993.1276. [DOI] [PubMed] [Google Scholar]
- Palese P., Compans R. W. Inhibition of influenza virus replication in tissue culture by 2-deoxy-2,3-dehydro-N-trifluoroacetylneuraminic acid (FANA): mechanism of action. J Gen Virol. 1976 Oct;33(1):159–163. doi: 10.1099/0022-1317-33-1-159. [DOI] [PubMed] [Google Scholar]
- Palese P., Tobita K., Ueda M., Compans R. W. Characterization of temperature sensitive influenza virus mutants defective in neuraminidase. Virology. 1974 Oct;61(2):397–410. doi: 10.1016/0042-6822(74)90276-1. [DOI] [PubMed] [Google Scholar]
- Schulman J. L., Palese P. Virulence factors of influenza A viruses: WSN virus neuraminidase required for plaque production in MDBK cells. J Virol. 1977 Oct;24(1):170–176. doi: 10.1128/jvi.24.1.170-176.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shibata S., Yamamoto-Goshima F., Maeno K., Hanaichi T., Fujita Y., Nakajima K., Imai M., Komatsu T., Sugiura S. Characterization of a temperature-sensitive influenza B virus mutant defective in neuraminidase. J Virol. 1993 Jun;67(6):3264–3273. doi: 10.1128/jvi.67.6.3264-3273.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tabak L. A., Levine M. J., Mandel I. D., Ellison S. A. Role of salivary mucins in the protection of the oral cavity. J Oral Pathol. 1982 Feb;11(1):1–17. doi: 10.1111/j.1600-0714.1982.tb00138.x. [DOI] [PubMed] [Google Scholar]
- Townsend A. R., Rothbard J., Gotch F. M., Bahadur G., Wraith D., McMichael A. J. The epitopes of influenza nucleoprotein recognized by cytotoxic T lymphocytes can be defined with short synthetic peptides. Cell. 1986 Mar 28;44(6):959–968. doi: 10.1016/0092-8674(86)90019-x. [DOI] [PubMed] [Google Scholar]
- Webster R. G., Laver W. G. Preparation and properties of antibody directed specifically against the neuraminidase of influenza virus. J Immunol. 1967 Jul;99(1):49–55. [PubMed] [Google Scholar]
- White J., Helenius A., Gething M. J. Haemagglutinin of influenza virus expressed from a cloned gene promotes membrane fusion. Nature. 1982 Dec 16;300(5893):658–659. doi: 10.1038/300658a0. [DOI] [PubMed] [Google Scholar]