Relationship between regulatory T cells and immune activation in human immunodeficiency virus-infected patients interrupting antiretroviral therapy

Laurence Weiss, Christophe Piketty, Lambert Assoumou, Céline Didier, Laure Caccavelli, Vladimira Donkova-Petrini, Yves Levy, Pierre-Marie Girard, Marianne Burgard, Jean-Paul Viard, Christine Rouzioux, Dominique Costagliola, ANRS 116 SALTO study group, M Bentata, P-M Girard, J-L Meynard, C Katlama, R Tubiana, A Simon, J-F Delfraissy, C Gougard, A Sobel, Y Levy, G Pialoux, J-P Viard, B Hoen, P Yeni, R Landmann, J A Gastaut, I Poizot-Martin, J M Ragnaut, P Morlat, T May, C Bazin, J M Lang, P Massip, J P Cassuto, P Choutet, Laurence Weiss, Christophe Piketty, Lambert Assoumou, Céline Didier, Laure Caccavelli, Vladimira Donkova-Petrini, Yves Levy, Pierre-Marie Girard, Marianne Burgard, Jean-Paul Viard, Christine Rouzioux, Dominique Costagliola, ANRS 116 SALTO study group, M Bentata, P-M Girard, J-L Meynard, C Katlama, R Tubiana, A Simon, J-F Delfraissy, C Gougard, A Sobel, Y Levy, G Pialoux, J-P Viard, B Hoen, P Yeni, R Landmann, J A Gastaut, I Poizot-Martin, J M Ragnaut, P Morlat, T May, C Bazin, J M Lang, P Massip, J P Cassuto, P Choutet

Abstract

Persistent immune activation plays a central role in driving Human Immunodeficiency Virus (HIV) disease progression. Whether CD4+CD25+ regulatory T cells (Tregs) are harmful by suppressing HIV-specific immune responses and/or beneficial through a decrease in immune activation remains debatable. We analysed the relationship between proportion and number of regulatory T cells (Tregs) and immune activation in HIV-infected patients interrupting an effective antiretroviral therapy (ART). Twenty-five patients were included in a substudy of a prospective multicenter trial of treatment interruption (TI) (ANRS 116). Proportions and numbers of Tregs and the proportion of activated CD4 and CD8 T cells were assessed at baseline and month 12 (M12) of TI. Specific anti-HIV CD4 and CD8 responses were investigated at baseline and M12. Non parametric univariate analyses and multivariate linear regression models were conducted. At baseline, the proportion of Tregs negatively correlated with the proportion of HLA-DR+CD8+T cells (r=-0.519). Following TI, the proportion of Tregs increased from 6.3% to 7.2% (p=0.029); absolute numbers of Tregs decreased. The increase in the proportion of HLA-DR+CD38+CD8+T cells was significantly related to the increase in proportion of Tregs (p=0.031). At M12, the proportion of Tregs did not negatively correlate with CD8 T-cell activation. Nevertheless, Tregs retain a suppressive function since depletion of Treg-containing CD4+CD25+ cells led to an increase in lymphoproliferative responses in most patients studied. Our data suggest that Tregs are efficient in controlling residual immune activation in patients with ART-mediated viral suppression. However, the insufficient increase in the proportion and/or the decrease in the absolute number of Tregs result in a failure to control immune activation following TI.

Trial registration: ClinicalTrials.gov NCT00118677.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1. The proportion of Tregs negatively…
Figure 1. The proportion of Tregs negatively correlated with CD8 T-cell activation at baseline but not at 12 months of treatment interruption (TI).
Panel A: Relationship between the percentages of CD8+ T cells expressing HLA-DR (x-axis) and the proportion of CD25+ CD127−/low Tregs among CD4+ T cells at baseline prior to TI (y-axis). Spearman's correlation coefficient R = −0.519, p = 0.008 Panel B: Relationship between the percentages of CD8+ T cells expressing HLA-DR and the proportion of CD25+ CD127−/low Tregs among CD4+ T cells after 12 months of TI (y-axis). Spearman's correlation coefficient R = 0.129; p = 0.539.
Figure 2. Proportion and absolute numbers of…
Figure 2. Proportion and absolute numbers of CD4+CD25+CD127−/low Tregs at baseline prior to treatment interruption (TI) and after 12 months off-treatment.
The expression of CD25 and CD127 was determined on thawed cryopreserved PBMC by 5-color flow cytometry after successively combining the FSC/SSC, CD3+ and CD4+ gates. Regulatory T cells were defined as CD25+ and CD127−/low.Results are expressed as percentages (panel A) and absolute numbers (panel B) of CD25+ CD127−/low Tregs among CD4+ T cells at baseline and after 12 months of TI (n = 25). The band near the middle of the box indicates the median, the bottom and the top of the boxes are the 25th and 75th percentile. Asterisks indicate a significant difference between M12 and baseline values. Representative stainings of PBMC from one patient at baseline (panel C) and at 12 months of TI (panel D) are shown.
Figure 3. The increase in the proportion…
Figure 3. The increase in the proportion of Tregs between baseline and M12 is independently related to the extent of generalized T-cell activation following treatment interruption.
Relationship between the change in the percentages of CD8+ T cells co-expressing HLA-DR and CD38 (x-axis) and the changes in the proportion of CD25+ CD127−/low Tregs among CD4+ T cells (y-axis) between baseline (prior to TI) and M12 of TI. Spearman's correlation coefficient R = 0.433; p = 0.031.
Figure 4. Predictors of the immunovirologic outcome…
Figure 4. Predictors of the immunovirologic outcome at month 12 of treatment interruption.
Baseline HIV-DNA levels predict the CD4 T-cell decline between baseline and M12 of TI. Relationship between the levels of HIV-DNA in PBMCs at baseline prior to TI (x-axis) and the change in CD4 cell counts between baseline and M12 of TI (y-axis). Spearman's correlation coefficient R = −0.578; p = 0.002.
Figure 5. The proportion of CD8+ T…
Figure 5. The proportion of CD8+ T cells co-expressing HLA-DR and CD38 at baseline predicts the increase in HIV-DNA levels between baseline and M12 of TI.
Relationship between the percentages of CD8+ T cells co-expressing HLA-DR and CD38 at baseline prior to TI (x-axis) and the change in HIV-DNA levels in PBMCs between baseline and M12 of TI (y-axis). The line corresponds to the polynomial regression line (order 2), the overall non parametric Spearman's correlation coefficient R is estimated as 0.552; p = 0.004, while it is 0.440 (p = 0.0357) after removing the two most extreme values.

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