Clinical information of patients with HIV-1.

*<p>VL is expressed as log viral copies/ml.</p><p>NC: Non controller.</p><p>VC: Viremic Controller.</p><p>EC: Elite Controller.</p

ADA enhances CD83, CD80 and CD86 expression on iDCs.

<p>iDCs, obtained as indicated in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0051287#s2" target="_blank">Materials and Methods</a>, were cultured for 48 h in medium in the absence (−ADA) or in the presence (+ADA) of 2 µM ADA or in the presence of maturating cocktail (mDCs). Expression of CD83 (A and B), CD80 (C and D) or CD86 (E and F) in the DCs gate was assessed by flow cytometry. In A, C and E, histogram overlays and the percentage of positive cells (A and B) or geometric mean (C) for a representative healthy donor and HIV-infected subject are shown. In B, D and F, values obtained from 16 to 19 healthy donors (circles) or 12 to 16 HIV-infected individuals (triangles) in the absence (open symbols) or in the presence (filled symbols) of 2 µM ADA are plotted. Each pair of linked symbols represents results from a particular individual. *<i>P</i><0.05; ***<i>P</i><0.001.</p

Effect of ADA on the iDCs viability.

<p>iDCs, obtained as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0051287#s2" target="_blank">Materials and Methods</a>, from healthy or HIV-infected donors were cultured during 48 h in the absence (−ADA) or in the presence (+ADA) of 2 µM ADA. Cell viability was assessed through DIOC₆ and propidium iodide (PI) staining and measured by flow cytometry. In (A), contour plots showing the percentage of viable (bright DIOC₆ and negative propidium iodide staining), apoptotic (low DIOC₆ and negative propidium iodide staining) and necrotic (low DIOC₆ and positive propidium iodide staining) populations from a representative healthy or HIV-infected donor are shown. Percentage of viable (B), apoptotic (C) and necrotic (D) DCs from 7 different healthy and 6 HIV-infected donors are shown.*<i>P</i><0.05.</p

Enzymatic and non-enzymatic activities are implicated on ADA-mediated effects.

<p>iDCs, obtained as indicated in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0051287#s2" target="_blank">Materials and Methods</a>, from 4 to 8 healthy donors were cultured for 48 h in medium in the absence (iDC) or in the presence of 2 µM ADA (+ADA) or 2 µM HgCl₂ inactivated ADA (ADA-Hg) or iDCs were pre-incubated with the mAb anti-CD26 TA5.9 and incubated with 2 µM ADA (ADA+TA5.9). In (A) and (B), the expression of CD83 and CD80 was assessed in the DCs gate by flow cytometry. In C) the indicated cytokines and chemokines were determined in the supernatants after 48 h of cell culture. Each pair of linked symbols represents results from a particular individual. Results are expressed as the ratio (in-fold) of the values obtained in the presence of ADA, ADA-Hg or ADA+TA5.9 versus untreated cells (iDCs, the reference value of 1 is represented by a dotted line). *<i>P</i><0.05; **<i>P</i><0.01 with respect to iDCs.</p

ADA enhances DCs immunogenicity in iDC-T-cell Allogeneic cocultures.

<p>In A and B, iDCs, obtained as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0051287#s2" target="_blank">Materials and Methods</a>, from a representative healthy donor were cultured during 48 h in medium in the absence of ADA (iDCs), in the presence of 2 µM ADA (iDCs+ADA) or in the presence of maturating cocktail (mDCs). DCs were washed and cocultured with allogeneic (upper contour plots) or autologous (lower contour plots) T-cells (1∶20 DCs:T-cells ratio). After 7 days, the percentage of CD4⁺ (A) and CD8⁺ (B) T-cell proliferation was assessed by flow cytometry using the CFSE method. In (C) percentages of CD4⁺ (circles) and CD8⁺ (squares) T-cell proliferation in the absence (open symbols) or the presence (filled symbols) of 2 µM ADA in allogeneic cocultures from 6 to 7 healthy donors are shown. Each pair of linked symbols represents results from a particular healthy donor. *<i>P</i><0.05. In (D) bars indicate IFN-γ, IL-4 and IL-17 levels in ADA-treated iDC co-culture supernatants. Results are the mean ± SD (pg/ml) of 3 independent experiments.</p

ADA increases cytokines and chemokines secretion.

<p>iDCs, obtained as indicated in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0051287#s2" target="_blank">Materials and Methods</a>, from 9 healthy (circles) and 8 HIV-infected (triangles) donors were cultured during 48 h in medium in the absence (iDCs) or in the presence of 2 µM ADA and the indicated cytokines and chemokines were determined in the supernatant as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0051287#s2" target="_blank">Materials and Methods</a>. Values are expressed as the ratio (in-fold) of cytokine or chemokine levels obtained in the presence of ADA versus levels obtained in the absence of ADA (iDCs, the reference value of 1 is represented by a dotted line). For each group, the median is indicated by a thick line. *<i>P</i><0.05; **<i>P</i><0.01 with respect to iDCs.</p