Research ArticleHIV

Delayed differentiation of potent effector CD8+ T cells reducing viremia and reservoir seeding in acute HIV infection

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Science Translational Medicine  15 Feb 2017:
Vol. 9, Issue 377, eaag1809
DOI: 10.1126/scitranslmed.aag1809
  • Fig. 1. HIV-specific CD8+ T cell expansion in acute HIV-1 infection.

    (A) Representative dot plot of Ki-67+Bcl-2lo CD8+ T cells from AHI week 0. (B) Percentages of Ki-67+Bcl-2lo cells in CD45RA CD8+ T cells from 14 HIV and 16 AHI 4G stage 1, 20 stage 2, and 27 stage 3 individuals. (C) Plasma viral load during the different stages of AHI week 0. (D) Correlation between plasma viral load and percentages of Ki-67+Bcl-2lo CD8+ T cells. (E) HLA-A*1101 NEF or EBV tetramer+ CD8+ T cells in two acutely infected individuals gated on total live peripheral blood lymphocytes (PBLs) (top). Overlay of Ki-67+Bcl-2lo expression on tetramer+ (red) and total CD45RA CD8+ T cells (blue) from the same participants (bottom). (F) Percentage of Ki-67+Bcl-2lo in HIV or EBV tetramer+ cells from two AHI stage 1, three stage 2, and six stage 3 participants. (G) HLA-A*1101 NEF or EBV tetramer+ CD8+ T cells gated on Ki-67+Bcl-2lo CD8+ T cells. Differences between groups were analyzed by Mann-Whitney tests. Associations between two variables (P and r) were analyzed by Spearman correlations. *P < 0.05; **P < 0.01; ****P < 0.0001.

  • Fig. 2. Fully differentiated effector HIV-specific CD8+ T cells in AHI contribute to viral load decrease after ART initiation.

    (A) Expression of perforin and T-bet in naïve CD8+ T cells and Ki-67+Bcl-2lo CD45RA CD8+ T cells from 14 HIV and 9 AHI stage 1, 9 stage 2, and 14 stage 3 individuals. (B) mRNA expression of granzyme B in CD38+HLA-DR+ CD8+ T cells from AHI stages 1/2 and 3 individuals. (C) Plasma viral load at week 2 in individuals who initiated ART in AHI stages 1 to 3. (D) Plasma viral load decrease between week 0 and week 2 after ART initiation in AHI stages 1 to 3 individuals. Differences between week 0 and week 2 were analyzed by Wilcoxon test. Slope differences are based on linear regression among stages and shown as P values. (E) Correlation between percentage of Ki-67+Bcl-2lo cells in CD45RA CD8+ T cells at week 0 and plasma viral load fold change (week 0/week 2). (F) Correlation between expression of effector molecules perforin and T-bet in Ki-67+Bcl-2lo CD8+ T cell at week 0 and plasma viral load fold change (week 0/week 2). Differences between groups were analyzed by Mann-Whitney tests. Associations between two variables (P and r) were analyzed by Spearman correlations. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

  • Fig. 3. Loss of cytokine secretion and survival potential of HIV-specific CD8+ T cells during AHI.

    (A) Expression of CD127 on CD38+HLA-DR+CD45RA CD8+ T cells from 12 AHI stage 1, 13 stage 2, and 14 stage 3 individuals. (B) mRNA expression of IL-7R and STAT5A in CD38+HLA-DR+ cells. (C) Interferon-γ (IFN-γ), TNF-α, and IL-2 production by CD38+HLA-DR+ cells from 9 AHI stage 1, 10 stage 2, and 11 stage 3 individuals after anti-CD3 and CD28-antibody stimulation. (D) Correlation between plasma viral load and percentages of IL-2+–secreting cells within CD38+HLA-DR+ cells. (E) Percentage of live nonapoptotic cell recovery of CD38+HLA-DR+ effector CD8+ T cells from AHI stages 1 to 3 individuals based on their absolute cell numbers after 24 hours in vitro. (F) Percentages of apoptotic annexin Vhi cells in ex vivo CD38+HLA-DR+ cells. (G) Correlation between plasma viral load and percentages of annexin Vhigh cells in CD38+HLA-DR+ CD8+ T cells. (H) Correlation between ex vivo apoptotic CD38+HLA-DR+ cell percentage and staining intensity of MitoTracker Orange CM-H2TMRos (mitochondria membrane potential activity, left) or CellROX Deep Red (total Ros level, right) from eight AHI stage 1, eight stage 2, and eight stage 3 individuals. Differences between groups were analyzed by Mann-Whitney tests. Associations between two variables (P and r) were analyzed by Spearman correlations. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

  • Fig. 4. Differential fate of HIV-specific CD8+ T cell 2 weeks after ART initiation in AHI.

    (A) Tetramer+ HLA-A*1101 Nef-specific CD8+ T cells in total live PBLs at week 0 and 2 weeks after ART initiation. (B) Tetramer+ HIV-specific CD8+ T cell absolute number per microliter based on total CD8+ T cell count for AHI stages 1 to 3 individuals. ND, not detected. (C) Percentage of Ki-67+Bcl-2lo cells in HIV-specific or EBV-specific tetramer+ CD8+ T cells from 7 AHI stage 1, 5 stage 2, and 11 stage 3 individuals at week 0 and week 2 on ART. (D) Percentage of Ki-67+ cells in CD8+ T cells from 14 stage 1, 20 stage 2, and 34 stage 3 individuals at week 0 and week 2. Differences between week 0 and week 2 were analyzed by Wilcoxon test. Slope differences are based on linear regression among stages and shown as P values. (E) Perforin expression in Ki-67+ and naïve CD8+ T cells at week 0 and week 2. Differences between groups were analyzed by Mann-Whitney tests. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

  • Fig. 5. HIV-specific CD8+ T cells persisting 2 weeks after ART initiation contribute to limited seeding and persistence of the HIV reservoir.

    (A) Correlation between percentage of Ki-67+ CD8+ T cells from 34 stage 3 individuals and plasma viral load at week 2. Open circle represents the detection limit. NS, not significant. (B) Total HIV DNA copies per 106 CD4+ T cells among AHI stages 1 to 3 individuals at week 2. For samples in which no positive cells were detected, the limit of detection based on cell input is plotted as an open symbol. (C) Correlation between percentage of Ki-67+ cells in total CD8+ T cells from 20 stage 2 individuals at week 2 and total HIV DNA copies per 106 CD4+ T cells at week 24. (D) Correlation between percentage of Ki-67+ CD8+ T cells at week 2 and total HIV DNA copies per 106 CD4+ T cells at weeks 24, 36, 48, 72, and 96 in AHI stage 3 individuals. (E) Correlation between percentage of Ki-67+CD4+ T cells at week 2 and total HIV DNA copies per 106 CD4+ T cells at week 24. Differences between groups were analyzed by Mann-Whitney tests. Associations between two variables (P and r) were analyzed by Spearman correlations. **P < 0.01; ****P < 0.0001.

  • Table 1. Clinical characteristic of study participants.
    HIV (n = 14)AHI 4G stage 1
    (n = 22)
    AHI 4G stage 2
    (n = 37)
    AHI 4G stage 3
    (n = 47)
    AHI 4G stagingNAT+
    4G/3G
    NAT+
    4G+/3G
    NAT+
    4G+/3G+
    WB or IND
    Days since HIV exposure, median [interquartile range (IQR)]NA14 (9–19)16 (13–20)19 (13–23)
    Age (years), mean (SD)32.7 (6.9)28.8 (8.3)28.4 (7.3)27.8 (6.7)
    Male/female ratio8/620/236/145/2
    CD4 count cells/μl in plasma, median (IQR)NA526.5 (334.3–575.5)304 (237.5–457)386 (295–506)
    CD8 count cells/μl in plasma, median (IQR)NA399.5 (262–482.3)240 (196–391)638 (425–1058)

Supplementary Materials

  • www.sciencetranslationalmedicine.org/cgi/content/full/9/377/eaag1809/DC1

    Materials and Methods

    Fig. S1. HIV-specific CD8+ T cell expansion in AHI.

    Fig. S2. CD4+ T cell activation in acute HIV-1 infection.

    Fig. S3. Characteristics of effector CD8+ T cells and plasma viral load during different stages of AHI.

    Fig. S4. Loss of memory potential and polyfunctionality of HIV-specific CD8+ T cell in AHI.

    Fig. S5. Mitochondrial function of HIV-specific CD8+ T cell during different stages of AHI.

    Fig. S6. HIV-specific CD8+ T cells 2 weeks after ART initiation during AHI.

    Fig. S7. Association between HIV-specific CD8+ T cells and HIV reservoir after ART initiation during AHI.

    Fig. S8. Association between activated CD4+ T cells and HIV reservoir after ART initiation during AHI.

    Table S1. Primary data.

  • Supplementary Material for:

    Delayed differentiation of potent effector CD8+ T cells reducing viremia and reservoir seeding in acute HIV infection

    Hiroshi Takata, Supranee Buranapraditkun, Cari Kessing, James L. K. Fletcher, Roshell Muir, Virginie Tardif, Pearline Cartwright, Claire Vandergeeten, Wendy Bakeman, Carmen N. Nichols, Suteeraporn Pinyakorn, Pokrath Hansasuta, Eugene Kroon, Thep Chalermchai, Robert O'Connell, Jerome Kim, Nittaya Phanuphak, Merlin L. Robb, Nelson L. Michael, Nicolas Chomont, Elias K. Haddad, Jintanat Ananworanich, Lydie Trautmann,* on behalf of the RV254/SEARCH010 and the RV304/SEARCH013 Study Groups

    *Corresponding author. Email: ltrautmann{at}hivresearch.org

    Published 15 February 2017, Sci. Transl. Med. 9, eaag1809 (2017)
    DOI: 10.1126/scitranslmed.aag1809

    This PDF file includes:

    • Materials and Methods
    • Fig. S1. HIV-specific CD8+ T cell expansion in AHI.
    • Fig. S2. CD4+ T cell activation in acute HIV-1 infection.
    • Fig. S3. Characteristics of effector CD8+ T cells and plasma viral load during different stages of AHI.
    • Fig. S4. Loss of memory potential and polyfunctionality of HIV-specific CD8+ T cell in AHI.
    • Fig. S5. Mitochondrial function of HIV-specific CD8+ T cell during different stages of AHI.
    • Fig. S6. HIV-specific CD8+ T cells 2 weeks after ART initiation during AHI.
    • Fig. S7. Association between HIV-specific CD8+ T cells and HIV reservoir after ART initiation during AHI.
    • Fig. S8. Association between activated CD4+ T cells and HIV reservoir after ART initiation during AHI.

    [Download PDF]

    Other Supplementary Material for this manuscript includes the following:

    • Table S1 (Microsoft Excel format). Primary data.

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