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Variability in a dominant block to SIV early reverse transcription in rhesus monkey cells predicts in vivo viral replication and time to death.

Rogers TF, Lim SY, Sundsvold TJ, Chan T, Hsu A, Letvin NL - Virol. J. (2010)

Bottom Line: While it has long been appreciated that there is considerable variability in host containment of HIV/SIV replication, the determinants of that variability are not fully understood.Previous studies demonstrated that the degree of permissivity of a macaque's peripheral blood mononuclear cells (PBMC) for infection with simian immunodeficiency virus (SIV) in vitro predicted that animal's peak plasma virus RNA levels following SIV infection in vivo.In a cohort of 15 unrelated Indian-origin rhesus monkeys, infectability of PBMC of individual animals with SIVsmE660, as defined by tissue culture infectious dose (TCID50), varied by more than 3 logs and was a stable phenotype over time.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Viral Pathogenesis, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02115, USA. thomas_rogers@hms.harvard.edu

ABSTRACT
While it has long been appreciated that there is considerable variability in host containment of HIV/SIV replication, the determinants of that variability are not fully understood. Previous studies demonstrated that the degree of permissivity of a macaque's peripheral blood mononuclear cells (PBMC) for infection with simian immunodeficiency virus (SIV) in vitro predicted that animal's peak plasma virus RNA levels following SIV infection in vivo. The present study was conducted to define the mechanisms underlying the variable intrinsic susceptibility of rhesus monkey PBMC to SIVsmE660 infection. In a cohort of 15 unrelated Indian-origin rhesus monkeys, infectability of PBMC of individual animals with SIVsmE660, as defined by tissue culture infectious dose (TCID50), varied by more than 3 logs and was a stable phenotype over time. Susceptibility of a monkey's PBMC to wild type SIVsmE660 infection correlated with the susceptibility of that monkey's PBMC to infection with VSV-G pseudotyped SIVsm543-GFP. Moreover, the permissivity of an individual monkey's PBMC for infection with this construct correlated with the permissivity of a B-lymphoblastoid cell line (B-LCL) generated from PBMC of the same animal. We found that the degree of intrinsic resistance of monkey B-LCL correlated with the copy number of early reverse transcription (ERT) SIV DNA. The resistance of monkey B-LCL to SIVsmE660 replication could be abrogated by preincubation of cells with the SIV virus-like particles (VLPs) and SIV resistance phenotype could be transferred to a SIV susceptible B-LCL through cell fusion. Finally, we observed a positive correlation between susceptibility of monkey B-LCL to SIV infection with a VSV-G pseudotyped SIV-GFP construct in vitro and both the peak plasma virus RNA levels in vivo and time to death following wild type SIV infection. These findings suggest that a dominant early RT restricting factor that can be saturated by SIV capsid may contribute to the variable resistance to SIV infection in rhesus monkey B-LCL and that this differential intrinsic susceptibility contributes to the clinical outcome of an SIV infection.

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B-LCL susceptibility to VSV-G pseudotyped SIV-GFP correlates with in vivo plasma viremia and time to death following rhesus monkey challenge with SIVmac251 or SHIV-89.6P. Positive correlation between % GFP+ B-LCL following VSV-G pseudotyped SIVsmE543-GFP in vitro infection of rhesus monkey B-LCL and (A) in vivo peak plasma viremia of monkeys (day 14 post-infection) as measured by RT assay, and (B) time to death following in vivo infection with SHIV-89.6P. Positive correlation between % GFP+ B-LCL following VSV-G pseudotyped SIVmac239-GFP in vitro infection of rhesus monkey B-LCL and (C) in vivo peak plasma virus RNA levels of monkeys (day 14 post-infection) and positive trend with (D) time to death following in vivo infection with SIVmac251.
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Figure 8: B-LCL susceptibility to VSV-G pseudotyped SIV-GFP correlates with in vivo plasma viremia and time to death following rhesus monkey challenge with SIVmac251 or SHIV-89.6P. Positive correlation between % GFP+ B-LCL following VSV-G pseudotyped SIVsmE543-GFP in vitro infection of rhesus monkey B-LCL and (A) in vivo peak plasma viremia of monkeys (day 14 post-infection) as measured by RT assay, and (B) time to death following in vivo infection with SHIV-89.6P. Positive correlation between % GFP+ B-LCL following VSV-G pseudotyped SIVmac239-GFP in vitro infection of rhesus monkey B-LCL and (C) in vivo peak plasma virus RNA levels of monkeys (day 14 post-infection) and positive trend with (D) time to death following in vivo infection with SIVmac251.

Mentions: To determine if this variable intracellular blockade of primate immunodeficiency virus replication impacts in vivo viral replication and clinical outcome in SIV-infected rhesus monkeys, we evaluated the permissivity of B-LCL generated from 14 monkeys for VSV-G pseudotyped SIVsm543-GFP infection. Then, following infection of the monkeys with SHIV89.6P, we assessed the correlation between the in vitro permissiveness of these B-LCL for replication of this virus and the in vivo peak plasma virus RNA levels and time to death following infection of rhesus monkeys with wild type virus (Fig. 8A, B). Additionally, we generated B-LCL from prechallenge PBMC of another cohort of rhesus monkeys that were infected with wild type SIVmac251 and assessed these B-LCL for susceptibility to SIVmac239-GFP replication. We observed a significant positive correlation and positive trend between in vitro B-LCL susceptibility to infection by these VSV-G pseudotyped SIV constructs and both peak viremia and time to death in the wild type SIV-infected monkeys, respectively (Fig. 8C, D). Monkeys whose B-LCL demonstrated a relative block to early RT exhibited a lower set point viral load following challenge in vivo with SIVmac251. These data indicate that in vitro permissivity of B-LCL for SIV replication is a reliable predictor of in vivo viral replication and clinical outcome.


Variability in a dominant block to SIV early reverse transcription in rhesus monkey cells predicts in vivo viral replication and time to death.

Rogers TF, Lim SY, Sundsvold TJ, Chan T, Hsu A, Letvin NL - Virol. J. (2010)

B-LCL susceptibility to VSV-G pseudotyped SIV-GFP correlates with in vivo plasma viremia and time to death following rhesus monkey challenge with SIVmac251 or SHIV-89.6P. Positive correlation between % GFP+ B-LCL following VSV-G pseudotyped SIVsmE543-GFP in vitro infection of rhesus monkey B-LCL and (A) in vivo peak plasma viremia of monkeys (day 14 post-infection) as measured by RT assay, and (B) time to death following in vivo infection with SHIV-89.6P. Positive correlation between % GFP+ B-LCL following VSV-G pseudotyped SIVmac239-GFP in vitro infection of rhesus monkey B-LCL and (C) in vivo peak plasma virus RNA levels of monkeys (day 14 post-infection) and positive trend with (D) time to death following in vivo infection with SIVmac251.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2874537&req=5

Figure 8: B-LCL susceptibility to VSV-G pseudotyped SIV-GFP correlates with in vivo plasma viremia and time to death following rhesus monkey challenge with SIVmac251 or SHIV-89.6P. Positive correlation between % GFP+ B-LCL following VSV-G pseudotyped SIVsmE543-GFP in vitro infection of rhesus monkey B-LCL and (A) in vivo peak plasma viremia of monkeys (day 14 post-infection) as measured by RT assay, and (B) time to death following in vivo infection with SHIV-89.6P. Positive correlation between % GFP+ B-LCL following VSV-G pseudotyped SIVmac239-GFP in vitro infection of rhesus monkey B-LCL and (C) in vivo peak plasma virus RNA levels of monkeys (day 14 post-infection) and positive trend with (D) time to death following in vivo infection with SIVmac251.
Mentions: To determine if this variable intracellular blockade of primate immunodeficiency virus replication impacts in vivo viral replication and clinical outcome in SIV-infected rhesus monkeys, we evaluated the permissivity of B-LCL generated from 14 monkeys for VSV-G pseudotyped SIVsm543-GFP infection. Then, following infection of the monkeys with SHIV89.6P, we assessed the correlation between the in vitro permissiveness of these B-LCL for replication of this virus and the in vivo peak plasma virus RNA levels and time to death following infection of rhesus monkeys with wild type virus (Fig. 8A, B). Additionally, we generated B-LCL from prechallenge PBMC of another cohort of rhesus monkeys that were infected with wild type SIVmac251 and assessed these B-LCL for susceptibility to SIVmac239-GFP replication. We observed a significant positive correlation and positive trend between in vitro B-LCL susceptibility to infection by these VSV-G pseudotyped SIV constructs and both peak viremia and time to death in the wild type SIV-infected monkeys, respectively (Fig. 8C, D). Monkeys whose B-LCL demonstrated a relative block to early RT exhibited a lower set point viral load following challenge in vivo with SIVmac251. These data indicate that in vitro permissivity of B-LCL for SIV replication is a reliable predictor of in vivo viral replication and clinical outcome.

Bottom Line: While it has long been appreciated that there is considerable variability in host containment of HIV/SIV replication, the determinants of that variability are not fully understood.Previous studies demonstrated that the degree of permissivity of a macaque's peripheral blood mononuclear cells (PBMC) for infection with simian immunodeficiency virus (SIV) in vitro predicted that animal's peak plasma virus RNA levels following SIV infection in vivo.In a cohort of 15 unrelated Indian-origin rhesus monkeys, infectability of PBMC of individual animals with SIVsmE660, as defined by tissue culture infectious dose (TCID50), varied by more than 3 logs and was a stable phenotype over time.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Viral Pathogenesis, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02115, USA. thomas_rogers@hms.harvard.edu

ABSTRACT
While it has long been appreciated that there is considerable variability in host containment of HIV/SIV replication, the determinants of that variability are not fully understood. Previous studies demonstrated that the degree of permissivity of a macaque's peripheral blood mononuclear cells (PBMC) for infection with simian immunodeficiency virus (SIV) in vitro predicted that animal's peak plasma virus RNA levels following SIV infection in vivo. The present study was conducted to define the mechanisms underlying the variable intrinsic susceptibility of rhesus monkey PBMC to SIVsmE660 infection. In a cohort of 15 unrelated Indian-origin rhesus monkeys, infectability of PBMC of individual animals with SIVsmE660, as defined by tissue culture infectious dose (TCID50), varied by more than 3 logs and was a stable phenotype over time. Susceptibility of a monkey's PBMC to wild type SIVsmE660 infection correlated with the susceptibility of that monkey's PBMC to infection with VSV-G pseudotyped SIVsm543-GFP. Moreover, the permissivity of an individual monkey's PBMC for infection with this construct correlated with the permissivity of a B-lymphoblastoid cell line (B-LCL) generated from PBMC of the same animal. We found that the degree of intrinsic resistance of monkey B-LCL correlated with the copy number of early reverse transcription (ERT) SIV DNA. The resistance of monkey B-LCL to SIVsmE660 replication could be abrogated by preincubation of cells with the SIV virus-like particles (VLPs) and SIV resistance phenotype could be transferred to a SIV susceptible B-LCL through cell fusion. Finally, we observed a positive correlation between susceptibility of monkey B-LCL to SIV infection with a VSV-G pseudotyped SIV-GFP construct in vitro and both the peak plasma virus RNA levels in vivo and time to death following wild type SIV infection. These findings suggest that a dominant early RT restricting factor that can be saturated by SIV capsid may contribute to the variable resistance to SIV infection in rhesus monkey B-LCL and that this differential intrinsic susceptibility contributes to the clinical outcome of an SIV infection.

Show MeSH
Related in: MedlinePlus