1887

Abstract

Nevirapine-resistant variants were generated by serial passages in MT-2 cells in the presence of increasing drug concentrations. In passage 5, mutations V106A, Y181C and G190A were detected in the global population, associated with a 100-fold susceptibility decrease. Sequence analysis of biological clones obtained from passage 5 and subsequent passages showed that single mutants, detected in first passages, were progressively replaced in passage 15 by double mutants, correlating with a 500-fold increase in phenotypic resistance. Fitness determination of single mutants confirmed that, in the presence of nevirapine, every variant was more fit than wild-type with a fitness order Y181C>V106A>G190A>wild-type. Unexpectedly, in the absence of the drug, the Y181C resistant mutant was more fit than wild-type, with a fitness gradient Y181C>wild-type >G106A⩾V190A. Using a molecular clone in which the Y181C mutation was introduced by mutagenesis, the greater fitness of the Y181C mutant was confirmed in new competition cultures. These data exemplify the role of resistance mutations on virus phenotype but also on virus evolution leading, occasionally, to resistant variants fitter than the wild-type in the absence of the drug.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-83-1-93
2002-01-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/jgv/83/1/0830093a.html?itemId=/content/journal/jgv/10.1099/0022-1317-83-1-93&mimeType=html&fmt=ahah

References

  1. Ayala F. J. 1971; Competition between species: frequency dependence. Science 171:820–824
    [Google Scholar]
  2. Balzarini J., Karlsson A., Perez-Perez M. J., Vrang L., Walbers J., Zhang H., Öberg B., Vandamme A. M., Camarasa M. J., De Clercq E. 1993; HIV-1-specific reverse transcriptase inhibitors show differential activity against HIV-1 mutant strains containing different amino acid substitutions in the reverse transcriptase. Virology 192:246–253
    [Google Scholar]
  3. Boom R., Sol C. J. A., Salimans M. M. M., Jansen C. L., Wertheim-van Dillen P. M. E., Van der Noordaa J. 1990; Rapid and simple method for purification of nucleic acids. Journal of Clinical Microbiology 28:495–503
    [Google Scholar]
  4. Borman A. M., Paulous S., Clavel F. 1996; Resistance of human immunodeficiency virus type 1 to protease inhibitors: selection of resistance mutations in the presence and absence of the drug. Journal of General Virology 77:419–426
    [Google Scholar]
  5. Boucher C. A. B., O’Sullivan E., Mulder J. W., Ramautarsing C., Kellam P., Darby G., Lange J. M., Goudsmit J., Larder B. A. 1992; Ordered appearance of zidovudine resistance mutations during treatment of 18 human immunodeficiency virus-positive subjects. Journal of Infectious Diseases 165:105–110
    [Google Scholar]
  6. Chao L. 1990; Fitness of RNA virus decreased by Muller’s ratchet. Nature 348:454–455
    [Google Scholar]
  7. Chesebro B., Wehrly K. 1988; Development of a sensitive quantitative focal assay for human immunodeficiency virus infectivity. Journal of Virology 62:3779–3788
    [Google Scholar]
  8. Chou T.-C., Talalay P. 1984; Quantitative analysis of dose–effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Advances in Enzyme Regulation 22:27–55
    [Google Scholar]
  9. Coffin J. M. 1995; HIV population dynamics in vivo: implications for genetic variation, pathogenesis, and therapy. Science 267:483–489
    [Google Scholar]
  10. Coffin J. M. 1996a; Plasma viral load, CD4+ cell counts, and HIV-1 production by cells. Science 271:670–671
    [Google Scholar]
  11. Coffin J. M. 1996b; Populations dynamics of HIV drug resistance. In Antiviral Drug Resistance pp 279–303 Edited by Richman D. D. Chichester, UK: John Wiley;
    [Google Scholar]
  12. De Clercq E. 1992; HIV inhibitors targeted at the reverse transcriptase. AIDS Research and Human Retroviruses 8:119–134
    [Google Scholar]
  13. de la Torre J. C., Holland J. J. 1990; RNA virus quasispecies populations can suppress vastly superior mutant progeny. Journal of Virology 64:6278–6281
    [Google Scholar]
  14. Delwart E. L., Sheppard H. W., Walker B. D., Goudsmit J., Mullins J. I. 1994; Human immunodeficiency virus type 1 evolution in vivo tracked by DNA heteroduplex mobility assays. Journal of Virology 68:6672–6683
    [Google Scholar]
  15. de Ronde A., van Dooren M., van Der Hoek L., Bouwhuis D., de Rooij E., van Gemen B., de Boer R., Goudsmit J. 2001; Establishment of new transmissible and drug-sensitive human immunodeficiency virus type 1 wild types due to transmission of nucleoside analogue-resistant virus. Journal of Virology 75:595–602
    [Google Scholar]
  16. Descamps D., Collin G., Letourneur F., Apetrei C., Damond F., Loussert-Ajaka I., Simon F., Saragosti S., Brun-Vézinet F. 1997; Susceptibility of human immunodeficiency virus type 1 group O isolates to antiretroviral agents: in vitro phenotypic and genotypic analyses. Journal of Virology 71:8893–8898
    [Google Scholar]
  17. Eigen M., Biebricher C. K. 1988; Sequence space and quasispecies distribution. In RNA Genetics pp 211–245 Edited by Domingo E., Holland J. J., Ahlquist P. Boca Raton, FL, USA: CRC Press;
    [Google Scholar]
  18. Escarmís C., Dávila M., Charpentier N., Bracho A., Moya A., Domingo E. 1996; Genetic lesions associated with Muller’s ratchet in an RNA virus. Journal of Molecular Biology 264:255–267
    [Google Scholar]
  19. Fischl M. A., Richman D. D., Griego M. H., Gottlieb M. S., Volberding P. A., Laskin O. L., Leedom J. M., Groopman J. E., Mildvan D., Schooley R. T., Jackson G. G., Durack D. T., King D., Group A. C. W. 1987; The efficacy of azidothymidine (AZT) in the treatment of patients with AIDS and AIDS-related complex. New England Journal of Medicine 317:185–191
    [Google Scholar]
  20. Gao Q., Gu Z., Parniak M. A., Li X., Wainberg M. A. 1992; In vitro selection of variants of human immunodeficiency virus type 1 resistant to 3′-azido-3′-deoxythymidine and 2′,3′-dideoxyinosine. Journal of Virology 66:12–19
    [Google Scholar]
  21. Gao Q., Gu Z., Salomon H., Nagai K., Parniak M. A., Wainberg M. A. 1994; Generation of multiple drug resistance by sequential in vitro passage of the human immunodeficiency virus type 1. Archives of Virology 136:111–122
    [Google Scholar]
  22. Goudsmit J., de Ronde A., Ho D. D., Perelson A. 1996; Human immunodeficiency virus fitness in vivo: calculations based on single zidovudine resistance mutation at codon 215 of reverse transcriptase. Journal of Virology 70:5662–5664
    [Google Scholar]
  23. Goudsmit J., de Ronde A., de Rooij E., de Boer R. 1997; Broad spectrum of in vivo fitness of human immunodeficiency virus type 1 subpopulations differing at reverse transcriptase codons 41 and 215. Journal of Virology 71:4479–4484
    [Google Scholar]
  24. Gu Z., Gao Q., Faust E. A., Wainberg M. A. 1995; Possible involvement of cell fusion and viral recombination in generation of human immunodeficiency virus variants that display dual resistance to AZT and 3TC. Journal of General Virology 76:2601–2605
    [Google Scholar]
  25. Harada S., Kobayashi N., Koyanagi Y., Yamamoto N. 1987; Clonal selection of human immunodeficiency virus (HIV): serological differences in the envelope antigens of the cloned viruses and HIV prototypes (HTLV-III B, LAV, and ARV). Virology 158:447–451
    [Google Scholar]
  26. Harrigan P. R., Kinghorn I., Bloor S., Kemp S. D., Najera I., Kohli A., Larder B. A. 1996; Significance of amino acid variation at human immunodeficiency virus type 1 reverse transcriptase residue 210 for zidovudine susceptibility. Journal of Virology 70:5930–5934
    [Google Scholar]
  27. Havlir D., Richman D. 1996; Non-nucleoside reverse transcriptase inhibitors – clinical aspects. In Antiviral Drug Resistance pp 241–260 Edited by Richman D. Chichester, UK: John Wiley;
    [Google Scholar]
  28. Havlir D. V., Eastman S., Gamst A., Richman D. D. 1996; Nevirapine-resistant human immunodeficiency virus: kinetics of replication and estimated prevalence in untreated patients. Journal of Virology 70:7894–7899
    [Google Scholar]
  29. Holland J. J., de la Torre J. C., Clarke D. K., Duarte E. 1991; Quantitation of relative fitness and great adaptability of clonal populations of RNA viruses. Journal of Virology 65:2960–2967
    [Google Scholar]
  30. Kellam P., Larder B. A. 1995; Retroviral recombination can lead to linkage of reverse transcriptase mutations that confer increased zidovudine resistance. Journal of Virology 69:669–674
    [Google Scholar]
  31. Kohlstaedt L. A., Wang J., Friedman J. M., Rice P. A., Steitz T. A. 1992; Crystal structure at 3·5 Å resolution of HIV-1 reverse transcriptase complexed with an inhibitor. Science 256:1783–1790
    [Google Scholar]
  32. Korber B., Kuiken C., Foley B., Hahn B., McCutchan F., Mellors J. W., Sodroski J. and others 1998 Human retroviruses and AIDS 1998: a compilation and analysis of nucleic acid and amino acid sequences Los Alamos, New Mexico: Theoretical Biology and Biophysics Group, Los Alamos National Laboratory;
    [Google Scholar]
  33. Kosalaraksa P., Kavlick M. F., Maroun V., Le R., Mitsuya H. 1999; Comparative fitness of multi-dideoxynucleoside-resistant human immunodeficiency virus type 1 (HIV-1) in an in vitro competitive HIV-1 replication assay. Journal of Virology 73:5356–5363
    [Google Scholar]
  34. Larder B. A., Purifoy D. J. M., Powell K. L., Darby G. 1987; Site-specific mutagenesis of AIDS virus reverse transcriptase. Nature 327:716–717
    [Google Scholar]
  35. Larder B. A., Darby G., Richman D. D. 1989; HIV with reduced sensitivity to zidovudine (AZT) isolated during prolonged therapy. Science 243:1731–1734
    [Google Scholar]
  36. Larder B. A., Chesebro B., Richman D. D. 1990; Susceptibilities of zidovudine-susceptible and -resistant human immunodeficiency virus isolates to antiviral agents determined by using a quantitative plaque reduction assay. Antimicrobial Agents and Chemotherapy 34:436–441
    [Google Scholar]
  37. Larder B. A., Coates K. E., Kemp S. D. 1991a; Zidovudine-resistant human immunodeficiency virus selected by passage in cell culture. Journal of Virology 65:5232–5236
    [Google Scholar]
  38. Larder B. A., Kellam P., Kemp S. D. 1991b; Zidovudine resistance predicted by direct detection of mutations in DNA from HIV-infected lymphocytes. AIDS 5:137–144
    [Google Scholar]
  39. López-Galíndez C., Guerra Romero L. 1997 Resistencia a los fármacos antirretrovirales Madrid: IDEPSA;
    [Google Scholar]
  40. Mascola J. R. 1999; Neutralization of HIV-1 infection of human peripheral blood mononuclear cells. In HIV Protocols (Methods in Molecular Medicine, 17) pp 317–322 Edited by Michael N. L., Kim J. H. Totowa: Human Press;
    [Google Scholar]
  41. Meyerhans A., Cheynier R., Albert J., Seth M., Kwok S., Sninsky J., Morfeldt-Manson L., Asjö B., Wain-Hobson S. 1989; Temporal fluctuations in HIV quasispecies in vivo are not reflected by sequential HIV isolations. Cell 58:901–910
    [Google Scholar]
  42. Miralles R., Gerrish P. J., Moya A., Elena S. F. 1999; Clonal interference and the evolution of RNA viruses. Science 285:1745–1747
    [Google Scholar]
  43. Mitsuya H., Weinhold K. J., Furman P. A., St Clair M. H., Lehrman S. N., Gallo R. C., Bolognesi D., Barry D. W., Broder S. 1985; 3′-Azido-3′-deoxythymidine (BW A509U): an antiviral agent that inhibits the infectivity and cytopathic effect of human T-lymphotropic virus type III/lymphadenopathy-associated virus in vitro. Proceedings of the National Academy of Sciences, USA 82:7096–7100
    [Google Scholar]
  44. Miyoshi I., Kubonishi I., Yoshimoto S., Akagi T., Ohtsuki Y., Shiraishi Y., Nagata K., Hinuma Y. 1981; Type C virus particles in a cord T-cell line derived by co-cultivating normal human cord leukocytes and human leukaemic T cells. Nature 294:770–771
    [Google Scholar]
  45. Molla A., Korneyeva M., Gao Q., Vasavanonda S., Schipper P. J., Mo H. M., Markowitz M., Chernyavsky T., Niu P., Lyons N., Hsu A., Granneman G. R., Ho D. D., Boucher C. A. B., Leonard J. M., Norbeck D. W., Kempf D. J. 1996; Ordered accumulation of mutations in HIV protease confers resistance to ritonavir. Nature Medicine 2:760–766
    [Google Scholar]
  46. Moutouh L., Corbeil J., Richman D. D. 1996; Recombination leads to the rapid emergence of HIV-1 dually resistant mutants under selective drug pressure. Proceedings of the National Academy of Sciences, USA 93:6106–6111
    [Google Scholar]
  47. Muller H. J. 1964; The relation of recombination to mutational advance. Mutation Research 1:2–29
    [Google Scholar]
  48. Nájera I., Richman D. D., Olivares I., Rojas J. M., Peinado M. A., Perucho M., Nájera R., López-Galindez C. 1994; Natural occurrence of drug resistance mutations in the reverse transcriptase of human immunodeficiency virus type 1 isolates. AIDS Research and Human Retroviruses 10:1479–1488
    [Google Scholar]
  49. Nájera I., Holguin A., Quiñones-Mateu M. E., Muñoz-Fernández M. A., Nájera R., López-Galíndez C., Domingo E. 1995; pol gene quasispecies of human immunodeficiency virus: mutations associated with drug resistance in virus from patients undergoing no drug therapy. Journal of Virology 69:23–31
    [Google Scholar]
  50. Nunberg J. H., Schleif W. A., Boots E. J., O’Brien J. A., Quintero J. C., Hoffman J. M., Emini E. A., Goldman M. E. 1991; Viral resistance to human immunodeficiency virus type 1-specific pyridinone reverse transcriptase inhibitors. Journal of Virology 65:4887–4892
    [Google Scholar]
  51. Olivares I., Shaw G., López-Galíndez C. 1997; Phenotypic switch in a Spanish HIV type 1 isolate on serial passage on MT-4 cells. AIDS Research and Human Retroviruses 13:979–984
    [Google Scholar]
  52. Olivares I., Casado Herrero C., Iglesias Ussel M. D., Dietrich U., López-Galíndez C. 1998; Complete sequence of an infectious molecular clone derived from a Spanish HIV type 1 isolate. AIDS Research and Human Retroviruses 14:1649–1651
    [Google Scholar]
  53. Perucho M., Goldfarb M., Shimizu K., Lama C., Fogh J., Wigler M. 1981; Human-tumor-derived cell lines contain common and different transforming genes. Cell 27:467–476
    [Google Scholar]
  54. Quiñones-Mateu M. E., Soriano V., Domingo E., Menéndez-Arias L. 1997; Characterization of the reverse transcriptase of a human immunodeficiency virus type 1 group O isolate. Virology 236:364–373
    [Google Scholar]
  55. Richman D. D., Shih C. K., Lowy I., Rose J., Prodanovich P., Goff S., Griffin J. 1991; Human immunodeficiency virus type 1 mutants resistant to nonnucleoside inhibitors of reverse transcriptase arise in tissue culture. Proceedings of the National Academy of Sciences, USA 88:11241–11245
    [Google Scholar]
  56. Richman D. D., Havlir D., Corbeil J., Looney D., Ignacio C., Spector S. A., Sullivan J., Cheeseman S., Barringer K., Pauletti D., Shih C. K., Myers M., Griffin J. 1994; Nevirapine resistance mutations of human immunodeficiency virus type 1 selected during therapy. Journal of Virology 68:1660–1666
    [Google Scholar]
  57. Sabino E., Pan L. Z., Cheng-Mayer C., Mayer A. 1994; Comparison of in vivo plasma and peripheral blood mononuclear cell HIV-1 quasispecies to short-term tissue culture isolates: an analysis of tat and C2–V3 env regions. AIDS 8:901–909
    [Google Scholar]
  58. Sánchez-Palomino S., Rojas J. M., Martínez M. A., Fenyö E. M., Nájera R., Domingo E., López-Galíndez C. 1993; Dilute passage promotes expression of genetic and phenotypic variants of human immunodeficiency virus type 1 in cell culture. Journal of Virology 67:2938–2943
    [Google Scholar]
  59. Shih C.-K., Rose J. M., Hansen G. L., Wu J. C., Bacolla A., Griffin J. A. 1991; Chimeric human immunodeficiency virus type 1/type 2 reverse transcriptases display reversed sensitivity to nonnucleoside analog inhibitors. Proceedings of the National Academy of Sciences, USA 88:9878–9882
    [Google Scholar]
  60. Smerdon S. J., Jäger J., Wang J., Kohlstaedt L. A., Chirino A. J., Friedman J. M., Rice P. A., Steitz T. A. 1994; Structure of the binding site for nonnucleoside inhibitors of the reverse transcriptase of human immunodeficiency virus type 1. Proceedings of the National Academy of Sciences, USA 91:3911–3915
    [Google Scholar]
  61. Spence R. A., Anderson K. S., Johnson K. A. 1996; HIV-1 reverse transcriptase resistance to nonnucleoside inhibitors. Biochemistry 35:1054–1063
    [Google Scholar]
  62. Tantillo C., Ding J., Jacobo-Molina A., Nanni R. G., Boyer P. L., Hughes S. H., Pauwles R., Andries K., Janssen P. A. J., Arnold E. 1994; Locations of anti-AIDS drug binding sites and resistance mutations in the three-dimensional structure of HIV-1 reverse transcriptase. Journal of Molecular Biology 243:369–387
    [Google Scholar]
  63. Wain-Hobson S. 1993; The fastest genome evolution ever described: HIV variation in situ. Current Opinion in Genetics & Development 3:878–883
    [Google Scholar]
  64. Yuste E., Sánchez-Palomino S., Casado C., Domingo E., López-Galíndez C. 1999; Drastic fitness loss in human immunodeficiency virus type 1 upon serial bottleneck events. Journal of Virology 73:2745–2751
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-83-1-93
Loading
/content/journal/jgv/10.1099/0022-1317-83-1-93
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error