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

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

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

Modulation of unique Vγ9Vδ2 T-cell subsets during HIV infection.

<p>Seven unique Vγ9Vδ2-cell subsets derived from all possible combinations of the three parameters were analyzed from the whole blood of healthy donors and compared to the HIV+ patients by flow cytometry after Picostim stimulation. The graph shows the absolute number of different unique Vγ9Vδ2T-cell subsets according to the particular combination of functions expressed. The results are shown as median and inter quartile range (box plot), and vertical lines show the minimum and maximum values. The Mann–Whitney U test was used to compare group medians. A p value less than 0.05 was considered statistically significant, ***p<0.001; **p<0,01; *p<0.05; HD = healthy donors, black bar; HIV+ = HIV-infected individuals, white bar.</p

CD107a-, IFNγ- and MIP 1β-producing Vγ9Vδ2 T-cell subsets decrease in HIV+ patients.

<p>According to the type of function, Vγ9Vδ2 T-cell response was analyzed in the healthy donors and compared to the HIV+ patients by flow cytometry after Picostim stimulation. (A) The absolute number of different Vγ9Vδ2 T-cell subsets according to the type of functions expressed (CD107a, IFNγ and MIP-1β). The results are shown as median and inter-quartile range (box plot), and vertical lines show the minimum and maximum values. (B) Summary pie chart; each sector of the pie chart is matched to the colored squares shown below the bar graphand shows the median percentages of responding Vγ9Vδ2 T-cells grouped according to the type of function analyzed. The Mann–Whitney U test was used to compare group medians and a permutation test based on χ² test to compare pie charts. A p value less than 0.05 was considered statistically significant, ***p<0.001; **p<0,01; *p<0.05; HD = healthy donors, black bar; HIV+ = HIV-infected individuals, white bar.</p

Clinical Characteristic of Patients at the Time of Blood Sampling.

<p>^apatients with >200 CD4 cells/mm³;</p><p>^bND: Not Detected;</p><p>^cpatients with <200 CD4 cells/mm³.</p><p>Clinical Characteristic of Patients at the Time of Blood Sampling.</p

Total and functional Vγ9Vδ2 T-cells correlate with CD4 cell count.

<p>Total (A) and functional (B) Vγ9Vδ2 T-cells from the HIV+ patients were correlated with CD4 cell count. Correlation statistics were analyzed using the Spearman R test and a p value less than 0.05 was considered statistically significant, ***p<0.001; **p<0,01; *p<0.05.</p

Responding Vγ9Vδ2-individuals decreases among HIV-infected patients who present lower Vγ9Vδ2 T-cell number than HD.

<p>Flow cytometer analyses were made to show (A) the absolute number of Vγ9Vδ2 T-cells from <i>ex vivo</i> HIV-infected patients (HIV+) and healthy donors (HD). The results are shown as median and interquartile range (box plot), and vertical lines show the minimum and maximum values; (B) the percentage of individuals among the HIV+ patients and HD having responding Vγ9Vδ2 T-cells after Picostim stimulation. Responding individuals, grey bar; Non-responding individuals, white bar. The Mann–Whitney U test was used to compare group medians. A p value less than 0.05 was considered statistically significant, ***p<0.001.</p