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Rapid seeding of the viral reservoir prior to SIV viraemia in rhesus monkeys.

Whitney JB, Hill AL, Sanisetty S, Penaloza-MacMaster P, Liu J, Shetty M, Parenteau L, Cabral C, Shields J, Blackmore S, Smith JY, Brinkman AL, Peter LE, Mathew SI, Smith KM, Borducchi EN, Rosenbloom DI, Lewis MG, Hattersley J, Li B, Hesselgesser J, Geleziunas R, Robb ML, Kim JH, Michael NL, Barouch DH - Nature (2014)

Bottom Line: Treatment with ART on day 3 blocked the emergence of viral RNA and proviral DNA in peripheral blood and also substantially reduced levels of proviral DNA in lymph nodes and gastrointestinal mucosa as compared with treatment at later time points.The time to viral rebound correlated with total viraemia during acute infection and with proviral DNA at the time of ART discontinuation.This strikingly early seeding of the refractory viral reservoir raises important new challenges for HIV-1 eradication strategies.

View Article: PubMed Central - PubMed

Affiliation: 1] Center for Virology and Vaccine Research, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215, USA [2] Ragon Institute of MGH, MIT and Harvard, Cambridge, Massachusetts 02139, USA.

ABSTRACT
The viral reservoir represents a critical challenge for human immunodeficiency virus type 1 (HIV-1) eradication strategies. However, it remains unclear when and where the viral reservoir is seeded during acute infection and the extent to which it is susceptible to early antiretroviral therapy (ART). Here we show that the viral reservoir is seeded rapidly after mucosal simian immunodeficiency virus (SIV) infection of rhesus monkeys and before systemic viraemia. We initiated suppressive ART in groups of monkeys on days 3, 7, 10 and 14 after intrarectal SIVMAC251 infection. Treatment with ART on day 3 blocked the emergence of viral RNA and proviral DNA in peripheral blood and also substantially reduced levels of proviral DNA in lymph nodes and gastrointestinal mucosa as compared with treatment at later time points. In addition, treatment on day 3 abrogated the induction of SIV-specific humoral and cellular immune responses. Nevertheless, after discontinuation of ART following 24 weeks of fully suppressive therapy, virus rebounded in all animals, although the monkeys that were treated on day 3 exhibited a delayed viral rebound as compared with those treated on days 7, 10 and 14. The time to viral rebound correlated with total viraemia during acute infection and with proviral DNA at the time of ART discontinuation. These data demonstrate that the viral reservoir is seeded rapidly after intrarectal SIV infection of rhesus monkeys, during the 'eclipse' phase, and before detectable viraemia. This strikingly early seeding of the refractory viral reservoir raises important new challenges for HIV-1 eradication strategies.

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SIV-specific humoral and cellular immune responses during ARTEnv-specific ELISA antibody titers at weeks 0, 4, 10, and 24 (a) and Env-, Pol-, and Gag-specific IFN-γ ELISPOT responses at weeks 0, 4, 10, and 20 in SIV-infected monkeys that initiated ART on days 3, 7, 10, and 14 of infection or with no ART (b). Mean responses are shown (N=4 animals per group). Error bars reflect standard errors.
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Figure 2: SIV-specific humoral and cellular immune responses during ARTEnv-specific ELISA antibody titers at weeks 0, 4, 10, and 24 (a) and Env-, Pol-, and Gag-specific IFN-γ ELISPOT responses at weeks 0, 4, 10, and 20 in SIV-infected monkeys that initiated ART on days 3, 7, 10, and 14 of infection or with no ART (b). Mean responses are shown (N=4 animals per group). Error bars reflect standard errors.

Mentions: We next assessed the development of SIV-specific humoral and cellular immune responses in these animals. Animals treated on day 3 following infection developed no detectable SIV Env-specific antibody responses by ELISA (Fig. 2a) and no detectable SIV Env-, Pol-, or Gag-specific T lymphocyte responses by IFN-γ ELISPOT assays (Fig. 2b) at weeks 4, 10, and 20 or 24 of infection. In contrast, animals treated on days 7, 10, and 14 developed detectable but lower SIV-specific humoral and cellular immune responses as compared with untreated controls, presumably as a result of reduced antigenic stimulus following ART initiation. Multiparameter intracellular cytokine staining (ICS) assays confirmed that SIV Gag-specific CD8+ and CD4+ T lymphocyte responses were undetectable in animals treated on day 3 and were lower in animals treated on days 7, 10, and 14 as compared with untreated controls (Extended Data Figs. 3–4). Gag-specific CD8+ and CD4+ T lymphocytes in animals treated on days 7, 10, and 14 also exhibited reduced immune activation and proliferation as measured by Ki67 expression as compared with untreated controls (Extended Data Fig. 4). These data demonstrate that initiation of ART on day 3 blocked the emergence of plasma viremia and abrogated the induction of SIV-specific humoral and cellular immune responses.


Rapid seeding of the viral reservoir prior to SIV viraemia in rhesus monkeys.

Whitney JB, Hill AL, Sanisetty S, Penaloza-MacMaster P, Liu J, Shetty M, Parenteau L, Cabral C, Shields J, Blackmore S, Smith JY, Brinkman AL, Peter LE, Mathew SI, Smith KM, Borducchi EN, Rosenbloom DI, Lewis MG, Hattersley J, Li B, Hesselgesser J, Geleziunas R, Robb ML, Kim JH, Michael NL, Barouch DH - Nature (2014)

SIV-specific humoral and cellular immune responses during ARTEnv-specific ELISA antibody titers at weeks 0, 4, 10, and 24 (a) and Env-, Pol-, and Gag-specific IFN-γ ELISPOT responses at weeks 0, 4, 10, and 20 in SIV-infected monkeys that initiated ART on days 3, 7, 10, and 14 of infection or with no ART (b). Mean responses are shown (N=4 animals per group). Error bars reflect standard errors.
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Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4126858&req=5

Figure 2: SIV-specific humoral and cellular immune responses during ARTEnv-specific ELISA antibody titers at weeks 0, 4, 10, and 24 (a) and Env-, Pol-, and Gag-specific IFN-γ ELISPOT responses at weeks 0, 4, 10, and 20 in SIV-infected monkeys that initiated ART on days 3, 7, 10, and 14 of infection or with no ART (b). Mean responses are shown (N=4 animals per group). Error bars reflect standard errors.
Mentions: We next assessed the development of SIV-specific humoral and cellular immune responses in these animals. Animals treated on day 3 following infection developed no detectable SIV Env-specific antibody responses by ELISA (Fig. 2a) and no detectable SIV Env-, Pol-, or Gag-specific T lymphocyte responses by IFN-γ ELISPOT assays (Fig. 2b) at weeks 4, 10, and 20 or 24 of infection. In contrast, animals treated on days 7, 10, and 14 developed detectable but lower SIV-specific humoral and cellular immune responses as compared with untreated controls, presumably as a result of reduced antigenic stimulus following ART initiation. Multiparameter intracellular cytokine staining (ICS) assays confirmed that SIV Gag-specific CD8+ and CD4+ T lymphocyte responses were undetectable in animals treated on day 3 and were lower in animals treated on days 7, 10, and 14 as compared with untreated controls (Extended Data Figs. 3–4). Gag-specific CD8+ and CD4+ T lymphocytes in animals treated on days 7, 10, and 14 also exhibited reduced immune activation and proliferation as measured by Ki67 expression as compared with untreated controls (Extended Data Fig. 4). These data demonstrate that initiation of ART on day 3 blocked the emergence of plasma viremia and abrogated the induction of SIV-specific humoral and cellular immune responses.

Bottom Line: Treatment with ART on day 3 blocked the emergence of viral RNA and proviral DNA in peripheral blood and also substantially reduced levels of proviral DNA in lymph nodes and gastrointestinal mucosa as compared with treatment at later time points.The time to viral rebound correlated with total viraemia during acute infection and with proviral DNA at the time of ART discontinuation.This strikingly early seeding of the refractory viral reservoir raises important new challenges for HIV-1 eradication strategies.

View Article: PubMed Central - PubMed

Affiliation: 1] Center for Virology and Vaccine Research, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215, USA [2] Ragon Institute of MGH, MIT and Harvard, Cambridge, Massachusetts 02139, USA.

ABSTRACT
The viral reservoir represents a critical challenge for human immunodeficiency virus type 1 (HIV-1) eradication strategies. However, it remains unclear when and where the viral reservoir is seeded during acute infection and the extent to which it is susceptible to early antiretroviral therapy (ART). Here we show that the viral reservoir is seeded rapidly after mucosal simian immunodeficiency virus (SIV) infection of rhesus monkeys and before systemic viraemia. We initiated suppressive ART in groups of monkeys on days 3, 7, 10 and 14 after intrarectal SIVMAC251 infection. Treatment with ART on day 3 blocked the emergence of viral RNA and proviral DNA in peripheral blood and also substantially reduced levels of proviral DNA in lymph nodes and gastrointestinal mucosa as compared with treatment at later time points. In addition, treatment on day 3 abrogated the induction of SIV-specific humoral and cellular immune responses. Nevertheless, after discontinuation of ART following 24 weeks of fully suppressive therapy, virus rebounded in all animals, although the monkeys that were treated on day 3 exhibited a delayed viral rebound as compared with those treated on days 7, 10 and 14. The time to viral rebound correlated with total viraemia during acute infection and with proviral DNA at the time of ART discontinuation. These data demonstrate that the viral reservoir is seeded rapidly after intrarectal SIV infection of rhesus monkeys, during the 'eclipse' phase, and before detectable viraemia. This strikingly early seeding of the refractory viral reservoir raises important new challenges for HIV-1 eradication strategies.

Show MeSH
Related in: MedlinePlus