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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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Positive correlation between rhesus monkey PBMC susceptibility to infection with wild type SIVsmE660 and a single cycle VSV-G pseudotyped SIVsmE660-GFP construct. (A) PBMC susceptibility to SIVsmE660 replication was defined by TCID50, and SIVsmE543-GFP infection was defined as % GFP+ PBMC following infection with VSV-G pseudotyped SIVsmE660-GFP determined using flow cytometric analysis. (B) Positive correlation between rhesus monkey PBMC susceptibility to SIVsmE660 infection, as defined by TCID50, and area under the curve (AUC) of % GFP+ B-LCL following infection with serial dilutions of VSV-G pseudotyped SIVsmE543-GFP. (C) Positive correlation between rhesus monkey PBMC susceptibility to SIVmac239 infection, as defined by TCID50, and area under the curve (AUC) of % GFP+ B-LCL following infection with serial dilutions of VSV-G pseudotyped SIVmac239-GFP.
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Figure 3: Positive correlation between rhesus monkey PBMC susceptibility to infection with wild type SIVsmE660 and a single cycle VSV-G pseudotyped SIVsmE660-GFP construct. (A) PBMC susceptibility to SIVsmE660 replication was defined by TCID50, and SIVsmE543-GFP infection was defined as % GFP+ PBMC following infection with VSV-G pseudotyped SIVsmE660-GFP determined using flow cytometric analysis. (B) Positive correlation between rhesus monkey PBMC susceptibility to SIVsmE660 infection, as defined by TCID50, and area under the curve (AUC) of % GFP+ B-LCL following infection with serial dilutions of VSV-G pseudotyped SIVsmE543-GFP. (C) Positive correlation between rhesus monkey PBMC susceptibility to SIVmac239 infection, as defined by TCID50, and area under the curve (AUC) of % GFP+ B-LCL following infection with serial dilutions of VSV-G pseudotyped SIVmac239-GFP.

Mentions: To determine if rhesus PBMC susceptibility to wild type SIVE660 is associated with susceptibility of these cells to a single-cycle pseudotyped SIV construct, rhesus monkey PBMC were infected with the VSV-G pseudotyped SIVSM543-GFP and analyzed by flow cytometry for % GFP-positive cells (Figure 3A). Rhesus monkey PBMC susceptibility to wild type SIVsmE660 virus was defined by TCID50, while susceptibility to the single cycle SIVsm543-GFP construct was defined as the % GFP+ PBMC 48 hrs post-infection. Rhesus monkey PBMC exhibited varied susceptibility to the single cycle VSV-G pseudotyped SIVSsm543-GFP, and a significant correlation was observed for each cell population between % SIVsm543-GFP+ cells and the TCID50 of these cells for wild type SIVsmE660 (Figure 3A). These data suggest that the differential permissivity of rhesus monkey PBMC to SIV replication is entry independent. Additionally, because the permissivity phenotype was manifested by the expression of GFP following a single cycle of replication, the relative blockage of viral replication must occur in the virus life cycle between early reverse transcription and post-integration viral expression. Moreover, the expression of this phenotype does not require multiple rounds of viral replication. Only one monkey's PBMC demonstrate a disparity between their permissivity to wild type virus and VSV-G pseudotyped SIVsm543-GFP.


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)

Positive correlation between rhesus monkey PBMC susceptibility to infection with wild type SIVsmE660 and a single cycle VSV-G pseudotyped SIVsmE660-GFP construct. (A) PBMC susceptibility to SIVsmE660 replication was defined by TCID50, and SIVsmE543-GFP infection was defined as % GFP+ PBMC following infection with VSV-G pseudotyped SIVsmE660-GFP determined using flow cytometric analysis. (B) Positive correlation between rhesus monkey PBMC susceptibility to SIVsmE660 infection, as defined by TCID50, and area under the curve (AUC) of % GFP+ B-LCL following infection with serial dilutions of VSV-G pseudotyped SIVsmE543-GFP. (C) Positive correlation between rhesus monkey PBMC susceptibility to SIVmac239 infection, as defined by TCID50, and area under the curve (AUC) of % GFP+ B-LCL following infection with serial dilutions of VSV-G pseudotyped SIVmac239-GFP.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 3: Positive correlation between rhesus monkey PBMC susceptibility to infection with wild type SIVsmE660 and a single cycle VSV-G pseudotyped SIVsmE660-GFP construct. (A) PBMC susceptibility to SIVsmE660 replication was defined by TCID50, and SIVsmE543-GFP infection was defined as % GFP+ PBMC following infection with VSV-G pseudotyped SIVsmE660-GFP determined using flow cytometric analysis. (B) Positive correlation between rhesus monkey PBMC susceptibility to SIVsmE660 infection, as defined by TCID50, and area under the curve (AUC) of % GFP+ B-LCL following infection with serial dilutions of VSV-G pseudotyped SIVsmE543-GFP. (C) Positive correlation between rhesus monkey PBMC susceptibility to SIVmac239 infection, as defined by TCID50, and area under the curve (AUC) of % GFP+ B-LCL following infection with serial dilutions of VSV-G pseudotyped SIVmac239-GFP.
Mentions: To determine if rhesus PBMC susceptibility to wild type SIVE660 is associated with susceptibility of these cells to a single-cycle pseudotyped SIV construct, rhesus monkey PBMC were infected with the VSV-G pseudotyped SIVSM543-GFP and analyzed by flow cytometry for % GFP-positive cells (Figure 3A). Rhesus monkey PBMC susceptibility to wild type SIVsmE660 virus was defined by TCID50, while susceptibility to the single cycle SIVsm543-GFP construct was defined as the % GFP+ PBMC 48 hrs post-infection. Rhesus monkey PBMC exhibited varied susceptibility to the single cycle VSV-G pseudotyped SIVSsm543-GFP, and a significant correlation was observed for each cell population between % SIVsm543-GFP+ cells and the TCID50 of these cells for wild type SIVsmE660 (Figure 3A). These data suggest that the differential permissivity of rhesus monkey PBMC to SIV replication is entry independent. Additionally, because the permissivity phenotype was manifested by the expression of GFP following a single cycle of replication, the relative blockage of viral replication must occur in the virus life cycle between early reverse transcription and post-integration viral expression. Moreover, the expression of this phenotype does not require multiple rounds of viral replication. Only one monkey's PBMC demonstrate a disparity between their permissivity to wild type virus and VSV-G pseudotyped SIVsm543-GFP.

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