Vγ9Vδ2 T cells fail to induce CD86 and HLA-DR up-regulation on HIV-infected MoDC.

<p>MoDC were infected with HIV_BAL and cultured with purified γδT cells for 5 days. Then, MoDC phenotype were evaluated by flow cytometry. (A) Representative histogram plots of one out of seven independent experiments showing CD86 expression on MoDC in the indicated conditions. (B) Induction of CD86 expression on MoDC by activated Vγ9Vδ2 T cells (fold of increase: IPP stimulated/not stimulated). (C) Representative histogram plots of one out of four independent experiments showing HLA-DR expression on MoDC in the indicated conditions. (D) HLA-DR expression on MoDC (mfi) in the indicated conditions. Results are shown as Box and Whiskers: the box encompasses the interquartile range of individual measurements, the horizontal bar-dividing line indicates the median value, and the whiskers represents maximum and minimum values.</p

Vγ9Vδ2 T cells do not inhibit HIV replication in MoDC.

<p>MoDC were infected with HIV_BAL and cultured with purified γδT cells stimulated or not stimulated with IPP. Before culture with γδT cells (DC+HIV T0), and after 5 days HIV p24 protein was tested in the supernatants by ELISA. Results from seven independent experiments are shown as Box and Whiskers: the box encompasses the interquartile range of individual measurements, the horizontal bar-dividing line indicates the median value, and the whiskers represents maximum and minimum values.</p

HIV-infected MoDC fail to down-regulate CCR5.

<p>MoDC were infected with HIV_BAL and cultured with purified γδT cells for 5 days. MoDC phenotype were evaluated by flow cytometry. (A) Representative histogram plots of one out of seven independent experiments showing CCR5 expression on MoDC in the indicated conditions. (B) Percentage of CCR5+ MoDC cultured with Vγ9Vδ2 T cells in the indicated conditions. Results are shown as Box and Whiskers: the box encompasses the interquartile range of individual measurements, the horizontal bar-dividing line indicates the median value, and the whiskers represents maximum and minimum values.</p

Effects of HIV-infected MoDC on Vγ9Vδ2 T cells proliferation.

<p>MoDC were infected with HIV_BAL and cultured with CFDA-SE labeled Vγ9Vδ2 T cells. After 5 days, Vγ9Vδ2 T cells proliferation and activation was evaluated by flow cytometry. (A) Representative histogram plots of one out of seven independent experiments showing Vγ9Vδ2 T cells proliferation. (B) CD69 expression on Vγ9Vδ2 T cells in the indicated conditions. Vγ9Vδ2 T cells labeled with CFDA-SE were stimulated with IPP in the presence of MoDC infected or not with HIV_BAL. After 5 days, Vγ9Vδ2 T cells proliferation was evaluated by flow cytometry. (C) Representative histogram plots of one out of seven independent experiments showing Vγ9Vδ2 T cells proliferation. (D) Percentage of proliferating Vγ9Vδ2 T cells upon IPP stimulation in the indicated conditions. (E) CD69 expression (mean fluorescence intensity, mfi) on IPP stimulated Vγ9Vδ2 T cells cultured with HIV infected or uninfected MoDC. Results are shown as Box and Whiskers: the box encompasses the interquartile range of individual measurements, the horizontal bar-dividing line indicates the median value, and the whiskers represents maximum and minimum values.</p

HIV-infected MoDC inhibit Vγ9Vδ2 T cells cytokines production.

<p>Vγ9Vδ2 T cells were stimulated with IPP in the presence of MoDC infected with HIV_BAL. After 5 days, cytokines released in the supernatants were evaluated by a multiplex immunoassay. (A) IFN-γ, (B) TNF-α, and (C) MIP1-β production, in seven independent experiments, are shown as Box and Whiskers: the box encompasses the interquartile range of individual measurements, the horizontal bar-dividing line indicates the median value, and the whiskers represents maximum and minimum values.</p

Frequency of HD and HIV+ presenting a T-cell response to different influenza A virus strains.

<p><b>* chi-square test:</b></p><p>**p = 0.0023.</p><p>Frequency of HD and HIV+ presenting a T-cell response to different influenza A virus strains.</p

HIV infection modulates Vδ1 and Vδ2 T-cells functionality.

<p>CD107a expression was analyzed in peripheral and in mucosal compartments in 3 P-HIV, 3 C-HIV patients and 3 HD by flow cytometry (Panel A). Basal IFNγ production by Vδ2 T-cells was analyzed in peripheral and in mucosal compartments in 4 P-HIV, 4 C-HIV patients and 4 HD by flow cytometry (Panel B). Data were considered significant with a P<0.05.</p

Vδ1 and Vδ2 T-cells in peripheral and mucosal compartments in HIV-patients and HD.

<p>Vδ1 and Vδ2 T-cells frequency was analyzed by flow cytometry in peripheral (Panels A-B) and in mucosal (Panels C-D) compartments of 15 P-HIV, 14 C-HIV patients and 35 HD. Data were considered significant with a p<0.05. Center line represents median; box represents interquartile range (IQR); whiskers represent range.</p

Correlation between humoral immune response to H3N2v strains.

<p>(A): The correlation between HAI titers to A/H3N2/Min/11/10 and to A/H3N2/Ind/08/11 is shown in HD. Pearson's r² and significance, as well as the calculated regression line. (B): The correlation between HAI titers to A/H3N2/Min/11/10 and to A/H3N2/Ind/08/11 is shown in HIV+. Pearson's r² and significance, as well as the calculated regression line.</p

Multi-parameter flow cytometric analysis of the functional profile of Vγ9Vδ2 T-cells.

<p>Representative dot plots of functional parameters (MIP-1β, CD107A, IFNγ panels B and D) and the respective Isotype control of antibodies (panels A and C) were shown in healthy donors (HD) and HIV-infected patients (HIV+) after stimulation with Picostim. For the gating strategy, after the lymphocyte population was gated sequentially, CD3+ and Vγ9Vδ2 T-cell events were defined, and gates for each respective function were made (panels B and D, light blue gates) using combinations that provided optimal separation. Single function gates were set based on the negative control (unstimulated) samples and were placed consistently across samples. We used the Boolean gate platform to create the full array of possible combinations, equating to 8 (2³) response patterns when testing three functions. Data are reported after background correction. Numbers on the light blue gates indicate the percentage of functional Vγ9Vδ2 T-cells among the total Vγ9Vδ2 T-cells (black boxes).</p