Abstract
In this study, we examined the peripheral blood (PB) central memory (TCM) CD4+ T cell subsets designated peripheral T follicular helper cells (pTfh cells) and non-pTfh cells to assess HIV permissiveness and persistence. Purified pTfh and non-pTfh cells from healthy HIV-negative donors were tested for HIV permissiveness using green fluorescent protein (GFP)-expressing HIV- 1NL4-3/Ba-L, followed by viral reactivation using beads coated with anti-CD3/anti-CD28 monoclonal antibodies. The role of pTfh cells in HIV persistence was analyzed in 12 chronically HIV-1 infected patients before and 48 weeks after initiation of raltegravir- containing combination antiretroviral therapy (cART). Total cellular HIV-1 DNA and episomes containing two copies of the viral long terminal repeat (2LTR circles) were analyzed in using droplet digital PCR in the purified pTfh and non-pTfh cells. Activation-inducible HIV p24 expression was determined by flow cytometry. Results indicate that pTfh cells, in particular PD1+ pTfh cells, showed greater permissiveness for HIV infection than non-pTfh cells. At week 48 on cART, HIV DNA levels were unchanged from pre-cART levels, although a significant decrease in 2LTR circles was observed in both cell subsets. Inducible HIV p24 expression was higher in pTfh cells than in non-pTfh cells, with the highest frequencies in the PD1+ CXCR3- pTfh cell subset. Frequencies of HLADR+ CD38+ activated CD4 T cells correlated with 2LTR circles in pTfh and non-pTfh cells at both time points and with p24+ cells at entry. In conclusion, among CD4 TCM cells in PB of aviremic patients on cART, pTfh cells, in particular the PD1+ CXCR3- subset, constitute a major HIV reservoir that is sustained by ongoing residual immune activation. The inducible HIV p24 assay is useful for monitoring HIV reservoirs in defined CD4 T cell subsets.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 2718-2728 |
| Number of pages | 11 |
| Journal | Journal of Virology |
| Volume | 90 |
| Issue number | 6 |
| DOIs | |
| State | Published - 2016 |
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ASJC Scopus subject areas
- Immunology
- Virology
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Peripheral T follicular helper cells are the major HIV reservoir within central memory CD4 T cells in peripheral blood from chronically HIV-infected individuals on combination antiretroviral therapy. / Pallikkuth, Suresh; Sharkey, Mark E; Babic, Dunja Z.; Gupta, Sachin; Stone, Geoffrey; Fischl, Margaret A; Stevenson, Mario; Pahwa, Savita G.
In: Journal of Virology, Vol. 90, No. 6, 2016, p. 2718-2728.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Peripheral T follicular helper cells are the major HIV reservoir within central memory CD4 T cells in peripheral blood from chronically HIV-infected individuals on combination antiretroviral therapy
AU - Pallikkuth, Suresh
AU - Sharkey, Mark E
AU - Babic, Dunja Z.
AU - Gupta, Sachin
AU - Stone, Geoffrey
AU - Fischl, Margaret A
AU - Stevenson, Mario
AU - Pahwa, Savita G
PY - 2016
Y1 - 2016
N2 - In this study, we examined the peripheral blood (PB) central memory (TCM) CD4+ T cell subsets designated peripheral T follicular helper cells (pTfh cells) and non-pTfh cells to assess HIV permissiveness and persistence. Purified pTfh and non-pTfh cells from healthy HIV-negative donors were tested for HIV permissiveness using green fluorescent protein (GFP)-expressing HIV- 1NL4-3/Ba-L, followed by viral reactivation using beads coated with anti-CD3/anti-CD28 monoclonal antibodies. The role of pTfh cells in HIV persistence was analyzed in 12 chronically HIV-1 infected patients before and 48 weeks after initiation of raltegravir- containing combination antiretroviral therapy (cART). Total cellular HIV-1 DNA and episomes containing two copies of the viral long terminal repeat (2LTR circles) were analyzed in using droplet digital PCR in the purified pTfh and non-pTfh cells. Activation-inducible HIV p24 expression was determined by flow cytometry. Results indicate that pTfh cells, in particular PD1+ pTfh cells, showed greater permissiveness for HIV infection than non-pTfh cells. At week 48 on cART, HIV DNA levels were unchanged from pre-cART levels, although a significant decrease in 2LTR circles was observed in both cell subsets. Inducible HIV p24 expression was higher in pTfh cells than in non-pTfh cells, with the highest frequencies in the PD1+ CXCR3- pTfh cell subset. Frequencies of HLADR+ CD38+ activated CD4 T cells correlated with 2LTR circles in pTfh and non-pTfh cells at both time points and with p24+ cells at entry. In conclusion, among CD4 TCM cells in PB of aviremic patients on cART, pTfh cells, in particular the PD1+ CXCR3- subset, constitute a major HIV reservoir that is sustained by ongoing residual immune activation. The inducible HIV p24 assay is useful for monitoring HIV reservoirs in defined CD4 T cell subsets.
AB - In this study, we examined the peripheral blood (PB) central memory (TCM) CD4+ T cell subsets designated peripheral T follicular helper cells (pTfh cells) and non-pTfh cells to assess HIV permissiveness and persistence. Purified pTfh and non-pTfh cells from healthy HIV-negative donors were tested for HIV permissiveness using green fluorescent protein (GFP)-expressing HIV- 1NL4-3/Ba-L, followed by viral reactivation using beads coated with anti-CD3/anti-CD28 monoclonal antibodies. The role of pTfh cells in HIV persistence was analyzed in 12 chronically HIV-1 infected patients before and 48 weeks after initiation of raltegravir- containing combination antiretroviral therapy (cART). Total cellular HIV-1 DNA and episomes containing two copies of the viral long terminal repeat (2LTR circles) were analyzed in using droplet digital PCR in the purified pTfh and non-pTfh cells. Activation-inducible HIV p24 expression was determined by flow cytometry. Results indicate that pTfh cells, in particular PD1+ pTfh cells, showed greater permissiveness for HIV infection than non-pTfh cells. At week 48 on cART, HIV DNA levels were unchanged from pre-cART levels, although a significant decrease in 2LTR circles was observed in both cell subsets. Inducible HIV p24 expression was higher in pTfh cells than in non-pTfh cells, with the highest frequencies in the PD1+ CXCR3- pTfh cell subset. Frequencies of HLADR+ CD38+ activated CD4 T cells correlated with 2LTR circles in pTfh and non-pTfh cells at both time points and with p24+ cells at entry. In conclusion, among CD4 TCM cells in PB of aviremic patients on cART, pTfh cells, in particular the PD1+ CXCR3- subset, constitute a major HIV reservoir that is sustained by ongoing residual immune activation. The inducible HIV p24 assay is useful for monitoring HIV reservoirs in defined CD4 T cell subsets.
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U2 - 10.1128/JVI.02883-15
DO - 10.1128/JVI.02883-15
M3 - Article
C2 - 26676775
AN - SCOPUS:84961161685
VL - 90
SP - 2718
EP - 2728
JO - Journal of Virology
JF - Journal of Virology
SN - 0022-538X
IS - 6
ER -