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eji2751-sup-0001-FigureS1.pdf310K

Supplementary Figure 1 Progressive gating strategy used to identify CD4+ T cells, CD8+ T cells and CD19+ B cells within the lymphocyte population and CD3- CD14+ cells within the monocyte population.

Supplementary Figure 2 Frequencies of CD4+ T cells, CD8+ T cells, CD19+ B cells and CD14+ monocytes that constitutively express IL-10 among ART-naive patients (n=25), ART-treated patients (n=20) and uninfected controls (n=5).

Supplementary Figure 3 Effect of depletion of HIV-1 gag-specific IL-10+ CD8+ T cells on HIV-1 gag-induced expression of HLA-DR and CD38 on CD4+ T cells.

Supplementary Figure 4 (A) CD38 expression on CD14+ monocytes from infected (p24 Ag+) and mock-infected PBMC (n = 4) after 3 and 5 days’ culture. CD8+ T cell-depleted PBMC from four HIV-negative subjects were activated for 3 days with phytohaemagglutinin and then infected with HIV-1BaL in the presence of IL-2, using a MOI to achieve infection of 5-10% CD4+ T cells, as indicated by expression of p24 antigen (p24 Ag). (B) Representative plots showing p24 Ag expression in monocytes from mock-infected PBMC cultures (left) and HIV-1BaL-infected PBMC cultures (right) after 3 days. (C) CD38 expression on CD14+ monocytes in mock-infected PBMC cultures (unexposed uninfected, UU), uninfected (p24 Agneg) monocytes in HIV-1BaL-infected PBMC (exposed uninfected, EU), or infected (p24 Ag+) monocytes in HIV-1BaL-infected PBMC (exposed infected, EI) (n = 6). Median values are indicated by horizontal bars.

Supplementary Figure 5 CD38 expression by monocytes in cultures where all CD8+ T cells were present (Undepleted), IL-10+ CD8+ T cells were depleted prior to co-culture of CD8+ and CD8neg fractions (“Depleted”) and where the CD8neg fraction was incubated with an IL-10R-blocking antibody prior to co-culture with undepleted CD8+ T cells (“Undepleted + αIL-10R”). Mean fluorescence intensity is expressed as arbitrary units. Three donors were tested; median values are indicated by horizontal bars.

eji2751-sup-0002-S1.pdf276KPeer review correspondence

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