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Persistence of viral genes in a variant of MDBK cell after productive replication of a mutant of influenza virus A/WSN

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Summary

The MDBK-R cell line is a variant of the MDBK cell line, which was derived by three consecutive high multiplicity superinfections of MDBK cells with AWBY-140 virus, a mutant of influenza virus A/WSN (H 1N 1). MDBK-R cells are permissive for productive replication of AWBY-140, but resist lysis by the virus and grew normally without producing infectious virus after replication of the mutant occurred there. By polymerase chain reaction (PCR), we demonstrated nucleotide sequences specific to all the 8 genes of AWBY-140 in MDBK-R cells which had been infected with the mutant at a high multiplicity and subsequently received 25 passages. This suggests that the genes of influenza virus mutant persisted in the dividing host cells for a long time after productive infection, when none of the cells was producing virus. We were also able to amplify the M gene related sequence of the mutant from both poly(A)+ and poly(A) fractions of the RNA extracted from the cells at 27th passage level by PCR, which suggests that the persisting genes were replicated and transcribed, but we failed to demonstrate any viral protein in the cells by Western blotting.

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References

  1. Buonagurio DA, Nakada S, Parvin JD, Krystal M, Palese P (1986) Evolution of human influenza A viruses over 50 years: rapid, uniform rate of changes in NS gene. Science 232: 980–982

    Google Scholar 

  2. Cane C, McLain L, Dimmock NJ (1987) Intracellular stability of the interfering activity of a defective interfering influenza virus in the absence of virus maturation. Virology 159: 259–264

    Google Scholar 

  3. Chien A, Edgar DB, Trela JM (1976) Deoxyribonucleic acid polymerase from the extreme thermophileThermus aquaticus. J Bacteriol 127: 1550–1557

    Google Scholar 

  4. De EK, Mayak DP (1980) Defective interfering influenza viruses and host cells: Establishment and maintenance of persistent influenza virus infection in MDBK and HeLa cells. J Virol 36: 847–859

    Google Scholar 

  5. Enami M, Luytjes M, Krystal M, Palese P (1990) Introduction of site specific mutations into the genome of influenza virus. Proc Natl Acad Sci USA 87: 3802–3805

    Google Scholar 

  6. Frielle DW, Huang DD, Youngner JR (1984) Persistent infection with influenza A viruses: evolution of virus mutants. Virology 138: 103–117

    Google Scholar 

  7. Gavrilov VI, Asher DM, Vyalushkina SD, Patushkina LS, Zmiev RG, Tumyan RG (1972) Persistent infection of continuous line of pig kidney cells with a variant of the WSN strain of influenza Ao virus. Proc Soc Exp Biol Med 140: 109–117

    Google Scholar 

  8. Hiti AL, Nayak DP (1982) Complete nucleotide sequence of the neuraminidase gene of human influenza virus A/WSN/33. J Virol 41: 730–734

    Google Scholar 

  9. Hiti AL, Davis AR, Nayak DP (1981) Complete sequence analysis shows that the hemagglutinin of the H O and H 2 subtypes of human influenza virus are closely related. Virology 111: 113–124

    Google Scholar 

  10. Kaptein JS, Nayak DP (1982) Complete nucleotide sequence of the polymerase 3 gene of human influenza virus A/WSN/33. J Virol 42: 55–63

    Google Scholar 

  11. Kilbourne ED (1987) Cytopathogenesis and cytopathology of influenza virus infection of cells in culture. In: Kilbourne ED (ed) Influenza. Plenum, New York, pp 89–110

    Google Scholar 

  12. Markushin S, Chiasi H, Sokolov A, Sinitsin B, Brown D, Klimov A, Nayak D (1988) Nucleotide sequence of RNA segment 7 and the predicted amino acid sequence of M 1 and M 2 proteins of FPV/Weybridge (H 7N 7) and WSN (H 1N 1) influenza viruses. Virus Res 10: 263–272

    Google Scholar 

  13. McBride LJ, Koepf SM, Gibbs RA, Salser W, Mayrand PE, Hunkapiller MW, Kronick MN (1989) Automated DNA sequencing methods involving polymerase chain reaction. Clin Chem 35: 2196–2201

    Google Scholar 

  14. Odagiri T, Tobita K (1990) Nucleotide sequence of the PA gene of influenza A/WSN/33 (H 1N 1). Nucleic Acid Res 18: 654

    Google Scholar 

  15. Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain termination inhibitors. Proc Natl Acad Sci USA 74: 5463–5467

    Google Scholar 

  16. Sivasubramanian N, Nayak DP (1982) Sequence analysis of the polymerase 1 gene and the secondary structure prediction of polymerase 1 protein of human influenza virus A/WSN/33. J Virol 44: 674–679

    Google Scholar 

  17. Smith LM, Sanders JZ, Kaiser RJ, Hughes P, Dodd C, Connell CR, Heiner C, Kent SBH, Hodd LE (1986) Fluorescence detection in automated DNA sequence analysis. Nature 321: 674–679

    Google Scholar 

  18. Tanaka T, Urabe M, Goto H, Tobita K (1984) Isolation and preliminary characterization of a highly cytoloytic influenza B virus mutant with an aberrant NS gene. Virology 135: 515–523

    Google Scholar 

  19. Tanaka T, Odagiri T, Tobita K (1988) Biological functions of the NS 1 protein of influenza B virus mutant which has a long carboxyl terminal deletion. Arch Virol 102: 173–185

    Google Scholar 

  20. Tobita K, Sugiura A, Enomoto C, Furuyama M (1975) Plaque assay and primary isolation of influenza A viruses in an established line of canine kidney cells (MDCK) in the presence of trypsin. Med Microbiol Immunol 162: 9–14

    Google Scholar 

  21. Tobita K, Tanaka T, Goto H, Feng S-Y (1983) Temperature-sensitive influenza A virus clones originated by a cross between A/Aichi/2/68 (H 3N 2) and B/Yamagata/1/73. Arch Virol 75: 17–27

    Google Scholar 

  22. Van Rompuy L, Min JW, Huylebroeck D, Devos R, Fiers W (1982) Complete nucleotide sequence of the nucleoprotein gene from the human influenza strain A/PR/8/34 (H 0N 1). Correction. Eur J Biochem 116: 645

    Google Scholar 

  23. Wang M, Webster RG (1990) Lack of persistence of influenza virus genetic information in ducks. Arch Virol 111: 263–267

    Google Scholar 

  24. Yanish-Perron C, Vieira J, Messing J (1985) Improved M 13 phage cloning vector and host strains: nucleotide sequence of the M 13mp 18 and pUc 19 vectors. Gene 33: 103–19

    Google Scholar 

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Urabe, M., Tanaka, T., Odagiri, T. et al. Persistence of viral genes in a variant of MDBK cell after productive replication of a mutant of influenza virus A/WSN. Archives of Virology 128, 97–110 (1993). https://doi.org/10.1007/BF01309791

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  • DOI: https://doi.org/10.1007/BF01309791

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