1887

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Volume 75, Issue 3

Molecular and biological characterization of simian immunodeficiency virus macaque strain 32H proviral clones containing size variants Free

Abstract

The proviral genome of the 32H reisolate of simian immunodeficiency of macaques (SIVmac32H) has been cloned and sequenced. Including both long terminal repeats, it is 10277 base pairs in length and contains open reading frames for all known SIV genes ( and ). This is the first report of an infectious SIVmac molecular clone which contains no premature termination codons. Three molecular clones of SIVmac32H have been constructed differing in sequence only within their last 1·2 kb. Two of the molecular clones, SIVmac32H(pJ5) and SIVmac32H (pC8), differ in the coding region by an in-frame deletion of four amino acids in pC8 and two conservative amino acid changes; other nucleotide changes in the 3′ LTR were not associated with known functionally critical motifs. The third clone, SIVmac32H(pB1), contains the last 1·2 kb of the SIVmac251 clone pBK28. The biological properties of virus produced after electroporation of these clones into C8166 cells has been assessed by infection of rhesus and cynomolgus macaques, time to seroconversion and by induction of cytopathic effects upon co-cultivation of infected rhesus peripheral blood lymphocytes with C8166 cells. The viruses obtained from these clones have identical growth kinetics but differ in their ability to persist in macaques. Macaques infected with pJ5 derived virus remain viraemic longer than macaques infected with pC8-derived virus. PCR analysis of circulating provirus indicates that the evolved over time in pJ5 virus-infected macaques, whereas late in infection in pC8 virus-infected macaques the gene remained invariant in sequence. These results support the observation that a deletion mutant of SIVmac239 lost its pathogenic potential and resulted in low-level viraemia when rhesus macaques were infected. Virus challenge pools for vaccine studies have been prepared for pJ5 using both human and monkey cell substrates and these stocks have been titrated both and . Virus has also been prepared from pC8 and titrated . This virus pool is being assessed as an attenuated live-virus vaccine in macaques. Since only virus originating from the SIVmac239 molecular clone is known to cause AIDSlike symptoms in rhesus macaques consistently, the SIVmac32H molecular clones should tell us more about which viral sequence features are important for the pathogenesis of AIDS.

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© The Journal of General Virology 1994
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1994-03-01
2024-04-20
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References

  1. Almond N., Jenkins A., Slade A., Heath A., Cranage M., Kitchin P. 1992; Population sequence analysis of a simian immunodeficiency virus (32H reisolate of SIVmac251): a virus stock used for international vaccine studies. AIDS Research and Human Retroviruses 8:77–88
     [Google Scholar]
  2. Binninger D., Ennen J., Bonn D., Norley S. G., Kurth R. 1991; Mutational analysis of the simian immunodeficiency virus SIVmac nef gene. Journal of Virology 65:5237–5343
     [Google Scholar]
  3. Bourgault I., Venet A., Levy J. P. 1992; Three epitopic peptides of the simian immunodeficiency virus Nef protein recognized by macaque cytotoxic T lymphocytes. Journal of Virology 66:750–756
     [Google Scholar]
  4. Burns D. P. W., Desrosiers R. C. 1991; Selection of genetic variants of simian immunodeficiency virus in persistently infected rhesus monkeys. Journal of Virology 65:1843–1854
     [Google Scholar]
  5. Carlson J. R., McGraw T. P., Keddie E., Yee J. L., Rosenthal A., Langlois A. J., Dickover R., Donovan R., Luciw P. A., Jennings M. B., Gardner M. B. 1990; Vaccine protection of rhesus macaques against simian immunodeficiency virus infection. AIDS Research and Human Retroviruses 6:1239–1246
     [Google Scholar]
  6. Chakrabarti L., Guyader M., Alizon M., Daniel M. D., Desrosiers R. C., Tiollais P., Sonigo P. 1987; Sequence of simian immunodeficiency virus from macaque and its relationship to other human and simian retroviruses. Nature; London: 328543–547
     [Google Scholar]
  7. Colombini S., Arya S. K., Reitz M. S., Jagodzinski L., Beaver B., Wong-Staal F. 1989; Structure of simian immunodeficiency virus regulatory genes. Proceedings of the National Academy of Sciences, U.S.A 86:4813–4817
     [Google Scholar]
  8. Cranage M. P., Cook N., Johnstone P., Greenaway P. J., Kitchin P. A., Stott E. J., Almond N., Baskerville A. 1990; SIV infection of rhesus macaques: in vivo titration of infectivity and development of an experimental vaccine. In Animal Models in AIDS pp. 103–113 Schellekens H., Horzinek M. C. Edited by Amsterdam: Elsevier;
     [Google Scholar]
  9. Cranage M. P., Stott E. J., Mills K. H. G., Ashworth A., Taffs F., Farrar G., Chan L., Dennis M., Putkonen P., Biberfeld G., Murphey-Corb M., Page M., Baskerville A., Kitchin P., Greenaway P. 1992; Vaccine studies with the 32H reisolate of SIVmac251: an overview. AIDS Research and Human Retroviruses 8:1479–1481
     [Google Scholar]
  10. Cullen B. R. 1991; Regulation of human immunodeficiency virus replication. Annual Review of Microbiology 45:219–250
     [Google Scholar]
  11. Daniel M. D., Letvin N. L., King N. W., Kannagi M., Sehgal P. K., Hunt R. D., Kanki P. J., Essex M., Desrosiers R. C. 1985; Isolation of a T-cell tropic HTLV-III-like retrovirus from macaques. Science 228:1201–1203
     [Google Scholar]
  12. Daniel M. D., Kirchhoff F., Czajak S. C., Sehgal P. K., Desrosiers R. C. 1992; Protective effects of a live attenuated SIV vaccine with a deletion in the nef gene. Science 258:1938–1941
     [Google Scholar]
  13. De Ronde A., Klaver B., Keulen W., Smit L., Goudsmit J. 1992; Natural HIV-1 NEF accelerates virus replication in primary lymphocytes. Virology 188:391–395
     [Google Scholar]
  14. Desrosiers R. C. 1992; HIV with multiple gene deletions as a live attenuated vaccine for AIDS. AIDS Research and Human Retroviruses 8:411–421
     [Google Scholar]
  15. Desrosiers R. C., Wyand M. S., Kodama T., Ringler D. J., Arthur L. O., Sehgal P. K., Letvin N. L., King N. W., Daniel M. D. 1989; Vaccine protection against simian immunodeficiency virus infection. Proceedings of the National Academy of Sciences, U.S.A 86:6353–6357
     [Google Scholar]
  16. Feichtinger H., Kaaya E., Putkonen P., Li S. -L., Ekman M., Gendelman R., Biberfeld G., Biberfeld P. 1992; Malignant lymphoma associated with human AIDS and with SIV-induced immunodeficiency in macaques. AIDS Research and Human Retroviruses 3:339–348
     [Google Scholar]
  17. Franchini G., Gurgo G., Guo H. -G., Gallo R. C., Collalti E., Fargnoli K. A., Hall L. F., Wong-Staal F., Reitz M. S. 1987; Sequence of simian immunodeficiency virus and its relationship to the human immunodeficiency virus. Nature; London: 328539–543
     [Google Scholar]
  18. Garcia J. V., Miller A. D. 1991; Serine phosphorylation-independent downregulation of cell-surface CD4 by Nef. Nature; London: 350508–511
     [Google Scholar]
  19. Green W. C. 1990; Regulation of HIV-1 gene expression. Annual Review of Immunology 8:453–475
     [Google Scholar]
  20. Guyader M., Emerman M., Sonigo P., Clavel F., Montagnier L., Alizon M. 1987; Genome organization and transactivation of human immunodeficiency virus type 2. Nature; London: 326662–669
     [Google Scholar]
  21. Hammes S. R., Dixon E. P., Malim M. H., Cullen B. R., Greene W. C. 1989; NEF protein of human immunodeficiency virus type 1: evidence against its role as a transcriptional inhibitor. Proceedings of the National Academy of Sciences, U.S.A 86:9549–9553
     [Google Scholar]
  22. Hirsch V., Riedel N., Mullins J. I. 1987; The genome organization of STLV-3 is similar to that of the AIDS virus except for a truncated transmembrane protein. Cell 49:307–319
     [Google Scholar]
  23. Hu S., Benveniste R. E. 1992; Protection of macaques against SIV infection by subunit vaccines of SIV envelope glycoprotein gpl60. Science 255:456–459
     [Google Scholar]
  24. Johnson P. R., Montefiori D. C., Goldstein S., Hamm T. E., Zhou J., Kitov S., Haigwood N. L., Misher L., London W. T., Gerin J. L., Allison A., Purcell R. H., Chanock R. M., Hirsch V. M. 1992; Inactivated whole-virus vaccine derived from a proviral DNA clone of simian immunodeficiency virus induces high levels of neutralizing antibodies and confers protection against heterologous challenge. Proceedings of the National Academy of Sciences, U.S.A 89:2175–2179
     [Google Scholar]
  25. Kent K. A., Gritz L., Stallard G., Cranage M. P., Collignon C., Thiriart C., Corcoran T., Silvera P., Stott E. J. 1991; Production of monoclonal antibodies to simian immunodeficiency virus envelope glycoproteins. AIDS 5:829–836
     [Google Scholar]
  26. Kestler H. W. III Ringler D. J., Mori K., Panicalli D. L., Sehgal P. K., Daniel M. D., Desrosiers R. C. 1991; Importance of the NEF gene for maintenance of high virus loads and for development of AIDS. Cell 65:651–662
     [Google Scholar]
  27. Kim S., Ikeuchi K., Byrn R., Groopman J., Baltimore D. 1989; Lack of a negative influence on viral growth by the nef gene of human immunodeficiency virus type 1. Proceedings of the National Academy of Sciences, U.S.A 86:9544–9548
     [Google Scholar]
  28. Kitchin P. A., Cranage M. P., Baskerville A., Greenaway P., Taffs F., Mills K. H. G., Page M., Almond N., Grief C., Corcoran T., Jenkins A., Ling C., Mahon B., McAlpine L., Silvera P., Szotyori Z., Stott E. J. 1990; Titration of SIVmac251(32H isolate) in cynomolgus macaques for use as a challenge in vaccination studies. In Animal Models in AIDS pp. 115–129 Schellekens H., Horzinek M. C. Edited by Amsterdam: Elsevier;
     [Google Scholar]
  29. Langlois A. J., Weinhold K. J., Matthews T. J., Greenberg M. L., Bolognesi D. P. 1992; Detection of anti-human cell antibodies in sera from macaques immunized with whole inactivated virus. AIDS Research and Human Retroviruses 8:1641–1652
     [Google Scholar]
  30. Little P. F. R. 1987; Choice and use of cosmid vectors. In DNA Cloning: A Practical Approach 3 pp. 19–42 Glover D. M. Edited by Oxford: IRL Press;
     [Google Scholar]
  31. Luria S., Chambers I., Berg P. 1991; Expression of the type 1 human immunodeficiency virus Nef protein in T cells prevents antigen receptor-mediated induction of interleukin 2 mRNA. Proceedings of the National Academy of Sciences, U.S.A 88:5326–5330
     [Google Scholar]
  32. Marthas M. L., Sutjipto S., Higgins J., Lohman B., Torten J., Luciw P. A., Marx P. A., Pedersen N. C. 1990; Immunization with a live, attenuated simian immunodeficiency virus (SIV) prevents early disease but not infection in rhesus macaques challenged with pathogenic SIV. Journal of Virology 64:3694–3700
     [Google Scholar]
  33. Mori K., Ringler D. J., Kodama T., Desrosiers R. C. 1992; Complex determinants of macrophage tropism in env of simian immunodeficiency virus. Journal of Virology 66:2067–2075
     [Google Scholar]
  34. Murphy-Corb M., Martin L. N., Davison-Fairburn B., Monte-Laro R. C., Miller M., West M., Ohkawa S., Baskin G. B., Zhang J. -Y., Putney S. D., Allison A. C., Eppstein D. A. 1989; A formalin-inactivated whole SIV vaccine confers protection in macaques. Science 246:1293–1297
     [Google Scholar]
  35. Naidu Y. M., Kestler H. W. III Li Y., Butler C. V., Silva D. P., Schmidt D. K., Troup C. D., Sehgal P. K., Sonigo P., Daniel M. D., Desrosiers R. C. 1988; Characterization of infectious molecular clones of simian immunodeficiency virus (SIVmac) and human immunodeficiency virus type 2: persistent infection of rhesus monkeys with molecular cloned SIVmac. Journal of Virology 62:4691–4696
     [Google Scholar]
  36. Rud E. W., Yon J. R., Larder B. A., Clarke B. E., Cook N., Cranage M. P. 1992; Infectious molecular clones of SIVmac32H: Nef deletion controls ability to reisolate virus from rhesus macaques. In Vaccines 92: Modern Approaches to New Vaccines Including Prevention of AIDS pp. 229–235 Brown F., Chanock R. M., Ginsberg H. S., Lerner R. A. Edited by New York: Cold Spring Harbor Laboratory;
     [Google Scholar]
  37. Rud E. W., Almond N., Ogilvie L., Kent K., Chan L., Page M., Kitchin P., Stott J., Cook N., Sharpe S., Ashworth T., Dennis M., Hall G., Cranage M. P., Clarke B. E. 1994; A naturally attenuated SIVmac32H vaccine or viral interference. In Vaccines 94: Modern Approaches to New Vaccines Including Prevention of AIDS Brown F., Chanock R. M., Ginsberg H. S., Norrby E. Edited by New York: Cold Spring Harbor Laboratory; in press
    [Google Scholar]
  38. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences, U.S.A 74:5463–5467
     [Google Scholar]
  39. Stott E. J., Chan W. L., Mills K. H. G., Page M., Taffs F., Cranage M., Greenaway P., Kitchin P. 1990; Preliminary report: protection of cynomolgus macaques against simian immunodeficiency virus by fixed infected cell vaccine. Lancet 336:1538–1541
     [Google Scholar]
  40. Stott E. J., Kitchin P. A., Page M., Flanagan B., Taffs L. F., Chan W. L., Mills K. H. G., Silvera P., Rodgers A. 1991; Anti-cell antibody in macaques. Nature; London: 353393
     [Google Scholar]
  41. Unger R. E., Marthas M. L., Pratt-Lowe E., Padrid P. A., Luciw P. A. 1992; The nef gene of simian immunodeficiency virus SIVmaclAl1. Journal of Virology 66:5432–5442
     [Google Scholar]
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Molecular and biological characterization of simian immunodeficiency virus macaque strain 32H proviral clones containing nef size variants
J. Gen. Virol. 75, 529 (1994); https://doi.org/10.1099/0022-1317-75-3-529
/content/journal/jgv/10.1099/0022-1317-75-3-529
/content/journal/jgv/10.1099/0022-1317-75-3-529
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