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Mouse adenovirus (MAV-1) expression in primary human endothelial cells and generation of a full-length infectious plasmid

Gene Therapy volume 6, pages 1291–1297 (1999)Cite this article

Abstract

Using RT-PCR, we show that mouse adenovirus type I (MAV-1) is capable of infecting and expressing in various cell types, specifically human endothelial cells. The capability of MAV-1 to infect and express in human endothelial cells makes it a potentially useful alternative to the use of human adenoviruses type 2/5 (Ad2/5) in virus-based gene therapy, although presently MAV-1 can only be produced at lower titers than Ad2/5. In this report, we present methods for the purification of MAV-1 DNA and use of this DNA along with a modified bacteria-based homologous recombination protocol to generate a full-length plasmid clone of MAV-1 DNA. Using various transfection procedures, we show that this plasmid MAV-1 DNA can generate plaques of MAV-1 virus, albeit at low efficiencies (about 0.2 p.f.u./μg DNA). Furthermore, the construction of an MAV-1 plasmid along with its capability to express in human cells justifies the full development of MAV-1 into a system of gene therapy.

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References

  1. Graham FL, Prevec L . Methods for construction of adenovirus vectors Mol Biotechnol 1995 3: 207–220

    Article  CAS  PubMed  Google Scholar 

  2. Kochanek S et al. A new adenoviral vector: replacement of all viral coding sequences with 28 kb of DNA independently expressing both full-length dystrophin and β-galactosidase Proc Natl Acad Sci USA 1996 93: 5731–5736

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Haecker SE et al. In vivo expression of full-length human dystrophin from adenoviral vectors deleted of all viral genes Hum Gene Ther 1996 7: 1907–1914

    Article  CAS  PubMed  Google Scholar 

  4. McGrory WJ, Bautista DS, Graham FL . A simple technique for the rescue of early region I mutations into infectious human adenovirus type 5 Virology 1988 163: 614–617

    Article  CAS  PubMed  Google Scholar 

  5. Chartier C et al. Efficient generation of recombinant adenovirus vectors by homologous recombination in Escherichia coli J Virol 1996 70: 4805–4810

    CAS  PubMed  PubMed Central  Google Scholar 

  6. Hartley JW, Rowe WP . A new mouse virus apparently related to the adenovirus group Virology 1960 11: 645–647

    Article  CAS  PubMed  Google Scholar 

  7. Gall J, Kass-Eisler A, Leinwand L, Falck-Pedersen E . Adenovirus type 5 and 7 capsid chimera: fiber replacement alters receptor tropism without affecting primary immune neutralization epitopes J Virol 1996 70: 2116–2123

    CAS  PubMed  PubMed Central  Google Scholar 

  8. Wickham TJ, Carrion ME, Kovesdi I . Targeting of adenovirus penton base to new receptors through replacement of its RGD motif with other receptor-specific peptide motifs Neuromusc Dis 1997 7: 284–298

    Article  Google Scholar 

  9. Douglas JT, Curiel DT . Strategies to accomplish targeted gene delivery to muscle cells employing tropism-modified adenoviral vectors Neuromusc Dis 1997 7: 284–298

    Article  CAS  PubMed  Google Scholar 

  10. Krasnykh VN, Mikheeva GV, Douglas JT, Curiel DT . Generation of recombinant adenovirus vectors with modified fibers for altering viral tropism J Virol 1996 70: 6839–6846

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Blailock ZR, Rabin ER, Melnick JL . Adenovirus endocarditis in mice Science 1967 157: 69–70

    Article  CAS  PubMed  Google Scholar 

  12. Heck FCJ, Sheldon WG, Gleiser CA . Pathogenesis of experimentally produced mouse adenovirus infection in mice Am J Vet Res 1972 33: 841–846

    PubMed  Google Scholar 

  13. Guida JD et al. Mouse adenovirus type 1 causes a fatal hemorrhagic encephalomyelitis in adult C57BL/6 but not BALB/c mice J Virol 1995 69: 7674–7681

    CAS  PubMed  PubMed Central  Google Scholar 

  14. Kring SC, King CS, Spindler KR . Susceptibility and signs associated with mouse adenovirus type 1 infection of adult outbred Swiss mice J Virol 1995 69: 8084–8088

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Engel JP . Viral upper respiratory infections Semin Resp Infect 1995 10: 3–13

    CAS  Google Scholar 

  16. Ketner G et al. Efficient manipulation of the human adenovirus genome as an infectious yeast artificial chromosome clone Proc Natl Acad Sci USA 1994 91: 6186–6190

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Hirt B . Selective extraction of polyoma DNA from infected mouse cell cultures J Mol Biol 1967 26: 365–369

    Article  CAS  PubMed  Google Scholar 

  18. Graham FL, Prevec L . Manipulation of adenovirus vectors. In: Murray EJ (ed) Gene Transfer and Expression Protocols Humana Press: Clifton, NJ 1991 pp 109–128

    Chapter  Google Scholar 

  19. Graham FL, van der Eb AJ . A new technique for the assay of infectivity of human adenovirus 5 DNA Virology 1973 52: 456–467

    Article  CAS  PubMed  Google Scholar 

  20. Larsen SH . Evolutionary variants of mouse adenovirus containing cellular DNA sequences Virology 1982 116: 573–580

    Article  CAS  PubMed  Google Scholar 

  21. Stoflet ES, Koeberl DD, Sarkar G, Sommer SS . Genomic amplification with transcript sequencing Science 1988 239: 491–494

    Article  CAS  PubMed  Google Scholar 

  22. Thornton SC, Mueller SN, Levine EM . Human endothelial cells: use of heparin in cloning and long-term serial cultivation Science 1983 222: 623–625

    Article  CAS  PubMed  Google Scholar 

  23. Gimbrone MAJ . Culture of vascular endothelium Prog Hemost Thromb 1976 3: 1–28

    PubMed  Google Scholar 

  24. Shinagawa M et al. A rapid and simple method for preparation of adenovirus DNA from infected cells Microbiol Immunol 1983 27: 817–822

    Article  CAS  PubMed  Google Scholar 

  25. Cox GA et al. Overexpression of dystrophin in transgenic mdx mice eliminates dystrophic symptoms without toxicity (see comments) Nature 1993 364: 725–729

    Article  CAS  PubMed  Google Scholar 

  26. Wang RF, Kushner SR . Construction of versatile low copy number vectors for cloning, sequencing and gene expression in Escherichia coli Gene 1991 100: 195–199

    Article  CAS  PubMed  Google Scholar 

  27. Wiegand RC, Beattie KL, Holloman WK, Radding CM . Uptake of homologous single-stranded fragments by superhelical DNA. III. The product and its enzymic conversion to a recombinant molecule J Mol Biol 1977 116: 805–824

    Article  CAS  PubMed  Google Scholar 

  28. Schaack J, Ho WY, Freimuth P, Shenk T . Adenovirus terminal protein mediates both nuclear matrix association and efficient transcription of adenovirus DNA Genes Dev 1990 4: 1197–1208

    Article  CAS  PubMed  Google Scholar 

  29. Miyake S et al. Efficient generation of recombinant adenoviruses using adenovirus DNA–terminal protein complex and a cosmid bearing the full-length virus genome Proc Natl Acad Sci USA 1996 93: 1320–1324

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Kring SC, Spindler KR . Lack of effect of mouse adenovirus type 1 infection on cell surface expression of major histocompatibility complex class I antigens J Virol 1996 70: 5495–5502

    CAS  PubMed  PubMed Central  Google Scholar 

  31. Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ . Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease Biochemistry 1979 18: 5294–5299

    Article  CAS  PubMed  Google Scholar 

  32. Meissner JD et al. Completion of the DNA sequence of mouse adenovirus type 1: sequence of E2B, L1, and L2 (18–51 map units) Virus Res 1997 51: 53–64

    Article  CAS  PubMed  Google Scholar 

  33. Ball AO, Williams ME, Spindler KR . Identification of mouse adenovirus type 1 early region 1: DNA sequence and a conserved transactivating function J Virol 1988 62: 3947–3957

    CAS  PubMed  PubMed Central  Google Scholar 

  34. Kusano K, Nakayama K, Nakayama H . Plasmid-mediated lethality and plasmid multimer formation in an Escherichia coli recBC sbcBC mutant. Involvement of RecF recombination pathway genes J Mol Biol 1989 209: 623–634

    Article  CAS  PubMed  Google Scholar 

  35. Hanahan D . Studies on transformation of Escherichia coli with plasmids J Mol Biol 1983 166: 557–580

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We thank Dr Christopher CW Hughes for providing Phec 97–54 primary umbilical endothelial cells. We thank Siddiqua Hirst and Mervin R Gutierrez for excellent technical assistance. This work was supported by a grant from the Muscular Dystrophy Association to LPV Additional facility support was from the Irvine Research Unit on Animal Viruses and the Viral Vector facility.

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Author notes

  1. J P Nery, G E Carney & K R Spindler

    Present address: Department of Molecular Microbiology and Immunology, Johns Hopkins University, Baltimore, MD, USA

  2. S Joseph

    Present address: School of Medicine, Tufts University, Boston, MA, USA

Authors and Affiliations

  1. Department of Molecular Biology and Biochemistry, University of California, Irvine, CA

    T T Nguyen, J P Nery, S Joseph, C E Rocha & L P Villarreal

  2. Department of Genetics, University of Georgia, Athens, GA, USA

    G E Carney & K R Spindler

  3. Department of Zoology, Oregon State University, Corvallis, OR, USA

    G E Carney

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  1. T T Nguyen
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Nguyen, T., Nery, J., Joseph, S. et al. Mouse adenovirus (MAV-1) expression in primary human endothelial cells and generation of a full-length infectious plasmid. Gene Ther 6, 1291–1297 (1999). https://doi.org/10.1038/sj.gt.3300949

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