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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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Proviral DNA in CD4+ T cell subpopulations during ART.Log proviral DNA (copies/106 CD4 T cells) in sorted naïve (N), transitional effector memory (TM), and central memory (CM) CD4 T cell subpopulations from PBMC and from genital, inguinal, iliac, para-aortic, axillary, and/or mesenteric lymph nodes obtained from two animals necropsied on day 3 and two animals necropsied on day 7 following mucosal SIVmac251 infection.
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Figure 10: Proviral DNA in CD4+ T cell subpopulations during ART.Log proviral DNA (copies/106 CD4 T cells) in sorted naïve (N), transitional effector memory (TM), and central memory (CM) CD4 T cell subpopulations from PBMC and from genital, inguinal, iliac, para-aortic, axillary, and/or mesenteric lymph nodes obtained from two animals necropsied on day 3 and two animals necropsied on day 7 following mucosal SIVmac251 infection.

Mentions: We next determined the impact of early ART on levels of proviral DNA14 in peripheral blood mononuclear cells (PBMC), lymph node mononuclear cells (LNMC), and gastrointestinal mucosa mononuclear cells (GMMC) over the course of 24 weeks of treatment with suppressive ART. In animals that initiated ART on day 3, there was a striking anatomic discordance with no proviral DNA detected in PBMC at any timepoint (<3 DNA copies/106 cells) (Fig. 3a). In contrast, clear but low levels of proviral DNA were detected in inguinal LNMC and in colorectal GMMC in these animals, although proviral DNA declined to undetectable or nearly undetectable levels in 3 of 4 of these animals by week 24. In monkeys treated with ART on days 7, 10, and 14, proviral DNA was readily detected in PBMC as well as in LNMC and GMMC (Fig. 3b–d). Moreover, in animals treated with ART on days 10 and 14, proviral DNA in LNMC appeared to stabilize by week 12 with minimal subsequent decline, consistent with a stable viral reservoir (Fig. 3c–d). In untreated animals, proviral DNA was markedly higher than in ART treated animals with minimal decline (Fig. 3e). Analysis of sorted cell subpopulations demonstrated that proviral DNA was found primarily in central memory and transitional memory CD4+ T lymphocytes in lymph nodes on day 3 and in both PBMC and lymph nodes on day 7 following SIV infection (Extended Data Fig. 5).


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)

Proviral DNA in CD4+ T cell subpopulations during ART.Log proviral DNA (copies/106 CD4 T cells) in sorted naïve (N), transitional effector memory (TM), and central memory (CM) CD4 T cell subpopulations from PBMC and from genital, inguinal, iliac, para-aortic, axillary, and/or mesenteric lymph nodes obtained from two animals necropsied on day 3 and two animals necropsied on day 7 following mucosal SIVmac251 infection.
© Copyright Policy
Related In: Results  -  Collection

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

Figure 10: Proviral DNA in CD4+ T cell subpopulations during ART.Log proviral DNA (copies/106 CD4 T cells) in sorted naïve (N), transitional effector memory (TM), and central memory (CM) CD4 T cell subpopulations from PBMC and from genital, inguinal, iliac, para-aortic, axillary, and/or mesenteric lymph nodes obtained from two animals necropsied on day 3 and two animals necropsied on day 7 following mucosal SIVmac251 infection.
Mentions: We next determined the impact of early ART on levels of proviral DNA14 in peripheral blood mononuclear cells (PBMC), lymph node mononuclear cells (LNMC), and gastrointestinal mucosa mononuclear cells (GMMC) over the course of 24 weeks of treatment with suppressive ART. In animals that initiated ART on day 3, there was a striking anatomic discordance with no proviral DNA detected in PBMC at any timepoint (<3 DNA copies/106 cells) (Fig. 3a). In contrast, clear but low levels of proviral DNA were detected in inguinal LNMC and in colorectal GMMC in these animals, although proviral DNA declined to undetectable or nearly undetectable levels in 3 of 4 of these animals by week 24. In monkeys treated with ART on days 7, 10, and 14, proviral DNA was readily detected in PBMC as well as in LNMC and GMMC (Fig. 3b–d). Moreover, in animals treated with ART on days 10 and 14, proviral DNA in LNMC appeared to stabilize by week 12 with minimal subsequent decline, consistent with a stable viral reservoir (Fig. 3c–d). In untreated animals, proviral DNA was markedly higher than in ART treated animals with minimal decline (Fig. 3e). Analysis of sorted cell subpopulations demonstrated that proviral DNA was found primarily in central memory and transitional memory CD4+ T lymphocytes in lymph nodes on day 3 and in both PBMC and lymph nodes on day 7 following SIV infection (Extended Data Fig. 5).

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