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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

Vδ2 T cell sorting strategy and purity.A) An example of the gating strategy to sort pre-enriched Vδ2 cells (panel 1) using a 2-step doublet discrimination (panels 2 and 3), showing that CD3+ Vδ2 cells (panel 4) lack expression of HLA-DR (panel 5). B) Dot plot examples showing lack of αβ-TCR (panel 6) and CD4 (panel 7) expression in the presort sample. C) Purity of the sorted cell population. After the sort an aliquot of sorted cells was acquired and analyzed. Plots show high purity of CD3+ Vδ2 cells (panel 8) and lack of HLA-DR expression (panel 9). Post-sort analysis shows lack of αβ-TCR and CD4 (panels 10 and 11).
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ppat.1005201.g001: Vδ2 T cell sorting strategy and purity.A) An example of the gating strategy to sort pre-enriched Vδ2 cells (panel 1) using a 2-step doublet discrimination (panels 2 and 3), showing that CD3+ Vδ2 cells (panel 4) lack expression of HLA-DR (panel 5). B) Dot plot examples showing lack of αβ-TCR (panel 6) and CD4 (panel 7) expression in the presort sample. C) Purity of the sorted cell population. After the sort an aliquot of sorted cells was acquired and analyzed. Plots show high purity of CD3+ Vδ2 cells (panel 8) and lack of HLA-DR expression (panel 9). Post-sort analysis shows lack of αβ-TCR and CD4 (panels 10 and 11).

Mentions: To ensure that other contaminating cells did not contribute to the recovery of HIV from isolated Vδ2 cells, we incubated freshly isolated patients’ PBMC with raltegravir and abacavir for 24 hours to avoid the possibility that de novo integration events could occur ex vivo after cell donation. γδ T cells were then enriched from PBMC using magnetic immunoaffinity beads, and non-activated (HLA-DR-) Vδ2 cells were further purified by FACS-sorting (Fig 1A and 1B). This process excluded αβTCR+ cells (classical CD4+ T cells) from pre-sort samples (Fig 1C), as detailed in Materials and Methods. To further confirm that Vδ2 cells were not already activated, aliquots of isolated Vδ2 cells were cultured in 5U/mL IL-2 prior to the addition of target cells in the viral outgrowth assay. HIV p24 measurements from these cultures were uniformly negative.


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)

Vδ2 T cell sorting strategy and purity.A) An example of the gating strategy to sort pre-enriched Vδ2 cells (panel 1) using a 2-step doublet discrimination (panels 2 and 3), showing that CD3+ Vδ2 cells (panel 4) lack expression of HLA-DR (panel 5). B) Dot plot examples showing lack of αβ-TCR (panel 6) and CD4 (panel 7) expression in the presort sample. C) Purity of the sorted cell population. After the sort an aliquot of sorted cells was acquired and analyzed. Plots show high purity of CD3+ Vδ2 cells (panel 8) and lack of HLA-DR expression (panel 9). Post-sort analysis shows lack of αβ-TCR and CD4 (panels 10 and 11).
© Copyright Policy
Related In: Results  -  Collection

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

ppat.1005201.g001: Vδ2 T cell sorting strategy and purity.A) An example of the gating strategy to sort pre-enriched Vδ2 cells (panel 1) using a 2-step doublet discrimination (panels 2 and 3), showing that CD3+ Vδ2 cells (panel 4) lack expression of HLA-DR (panel 5). B) Dot plot examples showing lack of αβ-TCR (panel 6) and CD4 (panel 7) expression in the presort sample. C) Purity of the sorted cell population. After the sort an aliquot of sorted cells was acquired and analyzed. Plots show high purity of CD3+ Vδ2 cells (panel 8) and lack of HLA-DR expression (panel 9). Post-sort analysis shows lack of αβ-TCR and CD4 (panels 10 and 11).
Mentions: To ensure that other contaminating cells did not contribute to the recovery of HIV from isolated Vδ2 cells, we incubated freshly isolated patients’ PBMC with raltegravir and abacavir for 24 hours to avoid the possibility that de novo integration events could occur ex vivo after cell donation. γδ T cells were then enriched from PBMC using magnetic immunoaffinity beads, and non-activated (HLA-DR-) Vδ2 cells were further purified by FACS-sorting (Fig 1A and 1B). This process excluded αβTCR+ cells (classical CD4+ T cells) from pre-sort samples (Fig 1C), as detailed in Materials and Methods. To further confirm that Vδ2 cells were not already activated, aliquots of isolated Vδ2 cells were cultured in 5U/mL IL-2 prior to the addition of target cells in the viral outgrowth assay. HIV p24 measurements from these cultures were uniformly negative.

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