1887

Abstract

A novel genetic approach for the control of virus replication was used for the design of a conditionally replicating human immunodeficiency virus (HIV) variant, HIV-rtTA. HIV-rtTA gene expression and virus replication are strictly dependent on the presence of a non-toxic effector molecule, doxycycline (dox), and thus can be turned on and off at will in a graded and reversible manner. The replication capacity, pathogenicity and genetic stability of this HIV-rtTA variant were evaluated in a humanized mouse model of haematopoiesis that harbours lymphoid and myeloid components of the human immune system (HIS). Infection of dox-fed BALB Rag/γc HIS (BRG-HIS) mice with HIV-rtTA led to the establishment of a productive infection without CD4 T-cell depletion. The virus did not show any sign of escape from dox control for up to 10 weeks after the onset of infection. No reversion towards a functional Tat–transactivating responsive (TAR) RNA element axis was observed, confirming the genetic stability of the HIV-rtTA variant . These results demonstrate the proof of concept that HIV-rtTA replicates efficiently . HIV-rtTA is a promising tool for fundamental research to study virus–host interactions in a controlled fashion.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.042796-0
2012-09-01
2024-12-14
Loading full text...

Full text loading...

/deliver/fulltext/jgv/93/9/2017.html?itemId=/content/journal/jgv/10.1099/vir.0.042796-0&mimeType=html&fmt=ahah

References

  1. An D. S., Poon B., Fang R. H. T., Weijer K., Blom B., Spits H., Chen I. S. Y., Uittenbogaart C. H. 2007; Use of a novel chimeric mouse model with a functionally active human immune system to study human immunodeficiency virus type 1 infection. Clin Vaccine Immunol 14:391–396 [View Article][PubMed]
    [Google Scholar]
  2. Baenziger S., Tussiwand R., Schlaepfer E., Mazzucchelli L., Heikenwalder M., Kurrer M. O., Behnke S., Frey J., Oxenius A.other authors 2006; Disseminated and sustained HIV infection in CD34+ cord blood cell-transplanted Rag2−/−γ c−/− mice. Proc Natl Acad Sci U S A 103:15951–15956 [View Article][PubMed]
    [Google Scholar]
  3. Becker P. D., Legrand N., van Geelen C. M., Noerder M., Huntington N. D., Lim A., Yasuda E., Diehl S. A., Scheeren F. A.other authors 2010; Generation of human antigen-specific monoclonal IgM antibodies using vaccinated “human immune system” mice. PLoS ONE 5:e13137 [View Article][PubMed]
    [Google Scholar]
  4. Berges B. K., Rowan M. R. 2011; The utility of the new generation of humanized mice to study HIV-1 infection: transmission, prevention, pathogenesis, and treatment. Retrovirology 8:65 [View Article][PubMed]
    [Google Scholar]
  5. Berges B. K., Wheat W. H., Palmer B. E., Connick E., Akkina R. 2006; HIV-1 infection and CD4 T cell depletion in the humanized Rag2−/−γc−/− (RAG-hu) mouse model. Retrovirology 3:76 [View Article][PubMed]
    [Google Scholar]
  6. Berkhout B., Marzio G., Verhoef K. 2001; Control over HIV-1 replication by an antibiotic; a novel vaccination strategy with a drug-dependent virus. Virus Res 82:103–108 [View Article][PubMed]
    [Google Scholar]
  7. Boom R., Sol C. J., Salimans M. M., Jansen C. L., Wertheim-van Dillen P. M., van der Noordaa J. 1990; Rapid and simple method for purification of nucleic acids. J Clin Microbiol 28:495–503[PubMed]
    [Google Scholar]
  8. Centlivre M., Klaver B., Berkhout B., Das A. T. 2008; Functional analysis of the complex trans-activating response element RNA structure in simian immunodeficiency virus. J Virol 82:9171–9178 [View Article][PubMed]
    [Google Scholar]
  9. Centlivre M., Zhou X., Pouw S. M., Weijer K., Kleibeuker W., Das A. T., Blom B., Seppen J., Berkhout B., Legrand N. 2010; Autoregulatory lentiviral vectors allow multiple cycles of doxycycline-inducible gene expression in human hematopoietic cells in vivo. Gene Ther 17:14–25 [View Article][PubMed]
    [Google Scholar]
  10. Chen Q., Khoury M., Chen J. 2009; Expression of human cytokines dramatically improves reconstitution of specific human-blood lineage cells in humanized mice. Proc Natl Acad Sci U S A 106:21783–21788 [View Article][PubMed]
    [Google Scholar]
  11. Das A. T., Verhoef K., Berkhout B. 2004a; A conditionally replicating virus as a novel approach toward an HIV vaccine. Methods Enzymol 388:359–379 [View Article][PubMed]
    [Google Scholar]
  12. Das A. T., Zhou X., Vink M., Klaver B., Verhoef K., Marzio G., Berkhout B. 2004b; Viral evolution as a tool to improve the tetracycline-regulated gene expression system. J Biol Chem 279:18776–18782 [View Article][PubMed]
    [Google Scholar]
  13. Das A. T., Harwig A., Vrolijk M. M., Berkhout B. 2007; The TAR hairpin of human immunodeficiency virus type 1 can be deleted when not required for Tat-mediated activation of transcription. J Virol 81:7742–7748 [View Article][PubMed]
    [Google Scholar]
  14. Denton P. W., García J. V. 2011; Humanized mouse models of HIV infection. AIDS Rev 13:135–148[PubMed]
    [Google Scholar]
  15. Emiliani S., Van Lint C., Fischle W., Paras P. Jr, Ott M., Brady J., Verdin E. 1996; A point mutation in the HIV-1 Tat responsive element is associated with postintegration latency. Proc Natl Acad Sci U S A 93:6377–6381 [View Article][PubMed]
    [Google Scholar]
  16. Gimeno R., Weijer K., Voordouw A., Uittenbogaart C. H., Legrand N., Alves N. L., Wijnands E., Blom B., Spits H. 2004; Monitoring the effect of gene silencing by RNA interference in human CD34+ cells injected into newborn RAG2−/− γc−/− mice: functional inactivation of p53 in developing T cells. Blood 104:3886–3893 [View Article][PubMed]
    [Google Scholar]
  17. Gorantla S., Sneller H., Walters L., Sharp J. G., Pirruccello S. J., West J. T., Wood C., Dewhurst S., Gendelman H. E., Poluektova L. 2007; Human immunodeficiency virus type 1 pathobiology studied in humanized BALB/c-Rag2−/−γc−/− mice. J Virol 81:2700–2712 [View Article][PubMed]
    [Google Scholar]
  18. Gorantla S., Makarov E., Finke-Dwyer J., Gebhart C. L., Domm W., Dewhurst S., Gendelman H. E., Poluektova L. Y. 2010; CD8+ cell depletion accelerates HIV-1 immunopathology in humanized mice. J Immunol 184:7082–7091 [View Article][PubMed]
    [Google Scholar]
  19. Huntington N. D., Alves N. L., Legrand N., Lim A., Strick-Marchand H., Mention J. J., Plet A., Weijer K., Jacques Y.other authors 2011; IL-15 transpresentation promotes both human T-cell reconstitution and T-cell-dependent antibody responses in vivo. Proc Natl Acad Sci U S A 108:6217–6222 [View Article][PubMed]
    [Google Scholar]
  20. Ince W. L., Zhang L., Jiang Q., Arrildt K., Su L., Swanstrom R. 2010; Evolution of the HIV-1 env gene in the Rag2−/− γC−/− humanized mouse model. J Virol 84:2740–2752 [View Article][PubMed]
    [Google Scholar]
  21. Jeeninga R. E., Westerhout E. M., van Gerven M. L., Berkhout B. 2008; HIV-1 latency in actively dividing human T cell lines. Retrovirology 5:37 [View Article][PubMed]
    [Google Scholar]
  22. Jiang Q., Zhang L., Wang R., Jeffrey J., Washburn M. L., Brouwer D., Barbour S., Kovalev G. I., Unutmaz D., Su L. 2008; FoxP3+CD4+ regulatory T cells play an important role in acute HIV-1 infection in humanized Rag2−/−γC−/− mice in vivo. Blood 112:2858–2868 [View Article][PubMed]
    [Google Scholar]
  23. Kiselyeva Y., Ito Y., Lima R. G., Grivel J.-C., Das A. T., Berkhout B., Margolis L. B. 2004; Depletion of CD4 T lymphocytes in human lymphoid tissue infected ex vivo with doxycycline-dependent HIV-1. Virology 328:1–6 [View Article][PubMed]
    [Google Scholar]
  24. Klausner R. D., Fauci A. S., Corey L., Nabel G. J., Gayle H., Berkley S., Haynes B. F., Baltimore D., Collins C.other authors 2003; The need for a global HIV vaccine enterprise. Science 300:2036–2039 [View Article][PubMed]
    [Google Scholar]
  25. Kleibeuker W., Zhou X., Centlivre M., Legrand N., Page M., Almond N., Berkhout B., Das A. T. 2009; A sensitive cell-based assay to measure the doxycycline concentration in biological samples. Hum Gene Ther 20:524–530 [View Article][PubMed]
    [Google Scholar]
  26. Kubota S., Pomerantz R. J. 1998; A cis-acting peptide signal in human immunodeficiency virus type I Rev which inhibits nuclear entry of small proteins. Oncogene 16:1851–1861 [View Article][PubMed]
    [Google Scholar]
  27. Legrand N., Huntington N. D., Nagasawa M., Bakker A. Q., Schotte R., Strick-Marchand H., de Geus S. J., Pouw S. M., Böhne M.other authors 2011; Functional CD47/signal regulatory protein alpha (SIRPα) interaction is required for optimal human T- and natural killer- (NK) cell homeostasis in vivo. Proc Natl Acad Sci U S A 108:13224–13229 [View Article][PubMed]
    [Google Scholar]
  28. Lepus C. M., Gibson T. F., Gerber S. A., Kawikova I., Szczepanik M., Hossain J., Ablamunits V., Kirkiles-Smith N., Herold K. C.other authors 2009; Comparison of human fetal liver, umbilical cord blood, and adult blood hematopoietic stem cell engraftment in NOD-scid/γc−/−, Balb/c-Rag1−/−γc−/−, and C.B-17-scid/bg immunodeficient mice. Hum Immunol 70:790–802 [View Article][PubMed]
    [Google Scholar]
  29. Marzio G., Verhoef K., Vink M., Berkhout B. 2001; In vitro evolution of a highly replicating, doxycycline-dependent HIV for applications in vaccine studies. Proc Natl Acad Sci U S A 98:6342–6347 [View Article][PubMed]
    [Google Scholar]
  30. Marzio G., Vink M., Verhoef K., de Ronde A., Berkhout B. 2002; Efficient human immunodeficiency virus replication requires a fine-tuned level of transcription. J Virol 76:3084–3088 [View Article][PubMed]
    [Google Scholar]
  31. O’Connell R. M., Balazs A. B., Rao D. S., Kivork C., Yang L., Baltimore D. 2010; Lentiviral vector delivery of human interleukin-7 (hIL-7) to human immune system (HIS) mice expands T lymphocyte populations. PLoS ONE 5:e12009 [View Article][PubMed]
    [Google Scholar]
  32. Pasternak A. O., Adema K. W., Bakker M., Jurriaans S., Berkhout B., Cornelissen M., Lukashov V. V. 2008; Highly sensitive methods based on seminested real-time reverse transcription-PCR for quantitation of human immunodeficiency virus type 1 unspliced and multiply spliced RNA and proviral DNA. J Clin Microbiol 46:2206–2211 [View Article][PubMed]
    [Google Scholar]
  33. Rongvaux A., Willinger T., Takizawa H., Rathinam C., Auerbach W., Murphy A. J., Valenzuela D. M., Yancopoulos G. D., Eynon E. E.other authors 2011; Human thrombopoietin knockin mice efficiently support human hematopoiesis in vivo. Proc Natl Acad Sci U S A 108:2378–2383 [View Article][PubMed]
    [Google Scholar]
  34. Sango K., Joseph A., Patel M., Osiecki K., Dutta M., Goldstein H. 2010; Highly active antiretroviral therapy potently suppresses HIV infection in humanized Rag2−/−γc−/− mice. AIDS Res Hum Retroviruses 26:735–746 [View Article][PubMed]
    [Google Scholar]
  35. Sato K., Izumi T., Misawa N., Kobayashi T., Yamashita Y., Ohmichi M., Ito M., Takaori-Kondo A., Koyanagi Y. 2010; Remarkable lethal G-to-A mutations in vif-proficient HIV-1 provirus by individual APOBEC3 proteins in humanized mice. J Virol 84:9546–9556 [View Article][PubMed]
    [Google Scholar]
  36. Strowig T., Rongvaux A., Rathinam C., Takizawa H., Borsotti C., Philbrick W., Eynon E. E., Manz M. G., Flavell R. A. 2011; Transgenic expression of human signal regulatory protein alpha in Rag2−/−γc−/− mice improves engraftment of human hematopoietic cells in humanized mice. Proc Natl Acad Sci U S A 108:13218–13223 [View Article][PubMed]
    [Google Scholar]
  37. ter Brake O., Legrand N., von Eije K. J., Centlivre M., Spits H., Weijer K., Blom B., Berkhout B. 2009; Evaluation of safety and efficacy of RNAi against HIV-1 in the human immune system (Rag-2−/−γc−/−) mouse model. Gene Ther 16:148–153 [View Article][PubMed]
    [Google Scholar]
  38. Traggiai E., Chicha L., Mazzucchelli L., Bronz L., Piffaretti J. C., Lanzavecchia A., Manz M. G. 2004; Development of a human adaptive immune system in cord blood cell-transplanted mice. Science 304:104–107 [View Article][PubMed]
    [Google Scholar]
  39. van der Kuyl A. C., Kozaczynska K., Ariën K. K., Gali Y., Balázs V. R., Dekker S. J., Zorgdrager F., Vanham G., Berkhout B., Cornelissen M. 2010; Analysis of infectious virus clones from two HIV-1 superinfection cases suggests that the primary strains have lower fitness. Retrovirology 7:60 [View Article][PubMed]
    [Google Scholar]
  40. van Lent A. U., Dontje W., Nagasawa M., Siamari R., Bakker A. Q., Pouw S. M., Maijoor K. A., Weijer K., Cornelissen J. J.other authors 2009; IL-7 enhances thymic human T cell development in “human immune system” Rag2−/−IL-2Rγc−/− mice without affecting peripheral T cell homeostasis. J Immunol 183:7645–7655 [View Article][PubMed]
    [Google Scholar]
  41. van Lent A. U., Centlivre M., Nagasawa M., Karrich J. J., Pouw S. M., Weijer K., Spits H., Blom B., Legrand N. 2010; In vivo modulation of gene expression by lentiviral transduction in “human immune system” Rag2−/−γc−/− mice. Methods Mol Biol 595:87–115 [View Article][PubMed]
    [Google Scholar]
  42. Verhoef K., Marzio G., Hillen W., Bujard H., Berkhout B. 2001; Strict control of human immunodeficiency virus type 1 replication by a genetic switch: Tet for Tat. J Virol 75:979–987 [View Article][PubMed]
    [Google Scholar]
  43. Vuyyuru R., Patton J., Manser T. 2011; Human immune system mice: current potential and limitations for translational research on human antibody responses. Immunol Res 51:257–266 [View Article][PubMed]
    [Google Scholar]
  44. Weijer K., Uittenbogaart C. H., Voordouw A., Couwenberg F., Seppen J., Blom B., Vyth-Dreese F. A., Spits H. 2002; Intrathymic and extrathymic development of human plasmacytoid dendritic cell precursors in vivo. Blood 99:2752–2759 [View Article][PubMed]
    [Google Scholar]
  45. Willinger T., Rongvaux A., Strowig T., Manz M. G., Flavell R. A. 2011; Improving human hemato-lymphoid-system mice by cytokine knock-in gene replacement. Trends Immunol 32:321–327 [View Article][PubMed]
    [Google Scholar]
  46. Yamada T., Iwamoto A. 2000; Comparison of proviral accessory genes between long-term nonprogressors and progressors of human immunodeficiency virus type 1 infection. Arch Virol 145:1021–1027 [View Article][PubMed]
    [Google Scholar]
  47. Zhang L., Kovalev G. I., Su L. 2007; HIV-1 infection and pathogenesis in a novel humanized mouse model. Blood 109:2978–2981[PubMed]
    [Google Scholar]
  48. Zhang L., Jiang Q., Li G., Jeffrey J., Kovalev G. I., Su L. 2011; Efficient infection, activation, and impairment of pDCs in the BM and peripheral lymphoid organs during early HIV-1 infection in humanized rag2−/−γ C−/− mice in vivo. Blood 117:6184–6192 [View Article][PubMed]
    [Google Scholar]
  49. Zhou X., Vink M., Berkhout B., Das A. T. 2006a; Modification of the Tet-On regulatory system prevents the conditional-live HIV-1 variant from losing doxycycline-control. Retrovirology 3:82 [View Article][PubMed]
    [Google Scholar]
  50. Zhou X., Vink M., Klaver B., Berkhout B., Das A. T. 2006b; Optimization of the Tet-On system for regulated gene expression through viral evolution. Gene Ther 13:1382–1390 [View Article][PubMed]
    [Google Scholar]
  51. Zhou X., Vink M., Klaver B., Verhoef K., Marzio G., Das A. T., Berkhout B. 2006c; The genetic stability of a conditional live HIV-1 variant can be improved by mutations in the Tet-On regulatory system that restrain evolution. J Biol Chem 281:17084–17091 [View Article][PubMed]
    [Google Scholar]
/content/journal/jgv/10.1099/vir.0.042796-0
Loading
/content/journal/jgv/10.1099/vir.0.042796-0
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF
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