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Peripheral Vγ9Vδ2 T Cells Are a Novel Reservoir of Latent HIV Infection.

Soriano-Sarabia N, Archin NM, Bateson R, Dahl NP, Crooks AM, Kuruc JD, Garrido C, Margolis DM - PLoS Pathog. (2015)

Bottom Line: Eradication of HIV infection will require the identification of all cellular reservoirs that harbor latent infection.We found that upregulation of the CD4 receptor may render primary Vδ2 cells target for HIV infection in vitro and we propose that HIV-induced immune activation may allow infection of γδ T cells in vivo.In 14 patients we recovered latent but replication-competent HIV from highly purified Vδ2 cells demonstrating that peripheral Vδ2 T cells are a previously unrecognized reservoir in which latent HIV infection is unexpectedly frequent.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, United States of America.

ABSTRACT
Eradication of HIV infection will require the identification of all cellular reservoirs that harbor latent infection. Despite low or lack of CD4 receptor expression on Vδ2 T cells, infection of these cells has previously been reported. We found that upregulation of the CD4 receptor may render primary Vδ2 cells target for HIV infection in vitro and we propose that HIV-induced immune activation may allow infection of γδ T cells in vivo. We assessed the presence of latent HIV infection by measurements of DNA and outgrowth assays within Vδ2 cells in 18 aviremic patients on long-standing antiretroviral therapy. In 14 patients we recovered latent but replication-competent HIV from highly purified Vδ2 cells demonstrating that peripheral Vδ2 T cells are a previously unrecognized reservoir in which latent HIV infection is unexpectedly frequent.

No MeSH data available.


Related in: MedlinePlus

Quantification of total HIV DNA levels.A) Total pol HIV copies were quantified by ddPCR within Vδ2 cells (n = 12), total resting CD4+ T cells (r-CD4) (n = 8) and unfractionated PBMC (n = 12) from HIV-1 suppressed patients treated in the acute HIV infection (AHI) or in the chronic HIV infection (CHI). Limit of quantitation (LOQ) was 50.6 copies/ 106 Vδ2 cells and 5.1 copies/ 106 r-CD4 cells and PBMC, and is depicted with a dotted line. Each color represents one patient. B) Pie charts reflecting the contribution of Vδ2 cells (purple) and r-CD4 cells (red) to the total HIV DNA+ PBMC in CHI patients (left pie) and AHI patients (right pie).
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ppat.1005201.g002: Quantification of total HIV DNA levels.A) Total pol HIV copies were quantified by ddPCR within Vδ2 cells (n = 12), total resting CD4+ T cells (r-CD4) (n = 8) and unfractionated PBMC (n = 12) from HIV-1 suppressed patients treated in the acute HIV infection (AHI) or in the chronic HIV infection (CHI). Limit of quantitation (LOQ) was 50.6 copies/ 106 Vδ2 cells and 5.1 copies/ 106 r-CD4 cells and PBMC, and is depicted with a dotted line. Each color represents one patient. B) Pie charts reflecting the contribution of Vδ2 cells (purple) and r-CD4 cells (red) to the total HIV DNA+ PBMC in CHI patients (left pie) and AHI patients (right pie).

Mentions: Total HIV DNA levels were then quantified in isolated Vδ2 cells, unfractionated PBMC and total resting CD4+ T (r-CD4) cells, when available (Fig 2A) in patients treated in AHI and CHI. As previously published in studies of other cell populations [22], DNA levels varied widely but interestingly, Vδ2 cells showed the highest level of pol HIV DNA copies per 106 Vδ2 cells (mean of 873.6 HIV copies/106 cells). Due to the low number of Vδ2 cells available for analysis the limit of quantitation of Vδ2 cells was 50.6 copies/106 cells, and 5.1 copies/106 cells for the other cell populations, where more cells could be analyzed. HIV DNA levels within Vδ2 cells were not statistically different between AHI and CHI-treated patients (p = 0.37). Within PBMC and r-CD4 cells HIV DNA levels were higher in CHI patients than in AHI patients, although this difference did not achieve statistical significance (p = 0.06 for PBMC and p = 0.65 for resting CD4+ T cells). We recovered an average of 638.6 HIV DNA copies/million γδ T cells from CHI patients, and an average of 1108.7 copies/ million from AHI patient. Conversely, we recovered an average of 30.3 HIV DNA copies/ million rCD4 cells from CHI patients and an average of 21.4 copies/ 106 rCD4 cells from AHI patients. We calculated the contribution of HIV DNA in Vδ2 cells and r-CD4 cells to the total HIV-DNA+ PBMC (Fig 2B) as follows: First, we calculated the total HIV DNA copies in each cell population by multiplying the average HIV copy number per million cells to the percentage of Vδ2 or r-CD4 cells present in total PBMC. Then, this total HIV copy number was divided by the total HIV copy number in PBMC to obtain the proportion of HIV corresponding to Vδ2or r-CD4 populations. Vδ2 cells contributed 1.6% and 8.1% to the total HIV DNA copy numbers in PBMC of CHI and AHI patients, respectively. Resting CD4 T cells contributed 4.9% to the total HIV DNA copy number in PBMC of CHI patients and 1.9% in AHI patients. None of the comparisons between cell types or type of patients were statistically different.


Peripheral Vγ9Vδ2 T Cells Are a Novel Reservoir of Latent HIV Infection.

Soriano-Sarabia N, Archin NM, Bateson R, Dahl NP, Crooks AM, Kuruc JD, Garrido C, Margolis DM - PLoS Pathog. (2015)

Quantification of total HIV DNA levels.A) Total pol HIV copies were quantified by ddPCR within Vδ2 cells (n = 12), total resting CD4+ T cells (r-CD4) (n = 8) and unfractionated PBMC (n = 12) from HIV-1 suppressed patients treated in the acute HIV infection (AHI) or in the chronic HIV infection (CHI). Limit of quantitation (LOQ) was 50.6 copies/ 106 Vδ2 cells and 5.1 copies/ 106 r-CD4 cells and PBMC, and is depicted with a dotted line. Each color represents one patient. B) Pie charts reflecting the contribution of Vδ2 cells (purple) and r-CD4 cells (red) to the total HIV DNA+ PBMC in CHI patients (left pie) and AHI patients (right pie).
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Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4608739&req=5

ppat.1005201.g002: Quantification of total HIV DNA levels.A) Total pol HIV copies were quantified by ddPCR within Vδ2 cells (n = 12), total resting CD4+ T cells (r-CD4) (n = 8) and unfractionated PBMC (n = 12) from HIV-1 suppressed patients treated in the acute HIV infection (AHI) or in the chronic HIV infection (CHI). Limit of quantitation (LOQ) was 50.6 copies/ 106 Vδ2 cells and 5.1 copies/ 106 r-CD4 cells and PBMC, and is depicted with a dotted line. Each color represents one patient. B) Pie charts reflecting the contribution of Vδ2 cells (purple) and r-CD4 cells (red) to the total HIV DNA+ PBMC in CHI patients (left pie) and AHI patients (right pie).
Mentions: Total HIV DNA levels were then quantified in isolated Vδ2 cells, unfractionated PBMC and total resting CD4+ T (r-CD4) cells, when available (Fig 2A) in patients treated in AHI and CHI. As previously published in studies of other cell populations [22], DNA levels varied widely but interestingly, Vδ2 cells showed the highest level of pol HIV DNA copies per 106 Vδ2 cells (mean of 873.6 HIV copies/106 cells). Due to the low number of Vδ2 cells available for analysis the limit of quantitation of Vδ2 cells was 50.6 copies/106 cells, and 5.1 copies/106 cells for the other cell populations, where more cells could be analyzed. HIV DNA levels within Vδ2 cells were not statistically different between AHI and CHI-treated patients (p = 0.37). Within PBMC and r-CD4 cells HIV DNA levels were higher in CHI patients than in AHI patients, although this difference did not achieve statistical significance (p = 0.06 for PBMC and p = 0.65 for resting CD4+ T cells). We recovered an average of 638.6 HIV DNA copies/million γδ T cells from CHI patients, and an average of 1108.7 copies/ million from AHI patient. Conversely, we recovered an average of 30.3 HIV DNA copies/ million rCD4 cells from CHI patients and an average of 21.4 copies/ 106 rCD4 cells from AHI patients. We calculated the contribution of HIV DNA in Vδ2 cells and r-CD4 cells to the total HIV-DNA+ PBMC (Fig 2B) as follows: First, we calculated the total HIV DNA copies in each cell population by multiplying the average HIV copy number per million cells to the percentage of Vδ2 or r-CD4 cells present in total PBMC. Then, this total HIV copy number was divided by the total HIV copy number in PBMC to obtain the proportion of HIV corresponding to Vδ2or r-CD4 populations. Vδ2 cells contributed 1.6% and 8.1% to the total HIV DNA copy numbers in PBMC of CHI and AHI patients, respectively. Resting CD4 T cells contributed 4.9% to the total HIV DNA copy number in PBMC of CHI patients and 1.9% in AHI patients. None of the comparisons between cell types or type of patients were statistically different.

Bottom Line: Eradication of HIV infection will require the identification of all cellular reservoirs that harbor latent infection.We found that upregulation of the CD4 receptor may render primary Vδ2 cells target for HIV infection in vitro and we propose that HIV-induced immune activation may allow infection of γδ T cells in vivo.In 14 patients we recovered latent but replication-competent HIV from highly purified Vδ2 cells demonstrating that peripheral Vδ2 T cells are a previously unrecognized reservoir in which latent HIV infection is unexpectedly frequent.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, United States of America.

ABSTRACT
Eradication of HIV infection will require the identification of all cellular reservoirs that harbor latent infection. Despite low or lack of CD4 receptor expression on Vδ2 T cells, infection of these cells has previously been reported. We found that upregulation of the CD4 receptor may render primary Vδ2 cells target for HIV infection in vitro and we propose that HIV-induced immune activation may allow infection of γδ T cells in vivo. We assessed the presence of latent HIV infection by measurements of DNA and outgrowth assays within Vδ2 cells in 18 aviremic patients on long-standing antiretroviral therapy. In 14 patients we recovered latent but replication-competent HIV from highly purified Vδ2 cells demonstrating that peripheral Vδ2 T cells are a previously unrecognized reservoir in which latent HIV infection is unexpectedly frequent.

No MeSH data available.


Related in: MedlinePlus