Functional profile and expression of co-stimulatory molecules and of co-inhibitory receptors of HIV-specific CD8 T-cell responses during acute and chronic HIV infections.

<p>Analysis of the functional profile (<b>A</b>), of the expression of CD27 and CD28 (<b>B</b>) and of the expression of 2B4, CD160 and PD-1 (<b>C</b>) in HIV-specific CD8 T cells in patients with acute (PHI-B), untreated chronic progressive (CP-B) or non-progressive (LTNP) HIV infection. Representative examples of the distinct flow cytometry panels are shown in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003423#ppat.1003423.s001" target="_blank">Fig. S1B</a>-D. Regarding the functional profile (<b>A</b>), although TNF-α was detected, analyses are restricted to the expression of IFN-γ, IL-2 and perforin for clarity. All possible combinations of the distinct markers are shown on the <i>x</i> axis, whereas the percentages of the distinct cell subsets within virus-specific CD8 T cells are shown on the <i>y</i> axis. The pie charts summarize the data, and each slice corresponds to a certain combination of molecules. Colors in the pie charts are based on the colored boxes at the bottom of the panel.</p

Increased CDR3 renewal of HIV-specific CD8 T cells following treatment interruption and association with functional avidity.

<p><b>A.</b> Percentage of CDR3 renewal of HIV-specific CD8 T cells before (under treatment) and after treatment interruption (TI). CDR3 diversity and renewal were determined as described <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003423#ppat.1003423-Miconnet1" target="_blank">[32]</a>. Example of TRBV usage and CDR3 size pattern analysis of B*0702-_GPGHKARVL-specific CD8 T cells in patient #1023 at week 18, 96 and 125 are shown in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003423#ppat.1003423.s003" target="_blank">Fig. S3</a>. <b>B.</b> Association between the percentage of CDR3 renewal and changes in the functional avidity of HIV-specific CD8 T cells.</p

Qualitative changes of HIV-specific CD8 T cells in patients experiencing a virus rebound following treatment interruption.

<p><b>A.</b> Longitudinal analysis of the CD4 T-cell counts and HIV viremia of 2 representative HIV-infected patients (#1017 and #1023) treated with ART since acute infection. Patient #1017 remained on ART, whereas patient #1023 spontaneously interrupted ART after 119 weeks (orange box). <b>B.</b> Functional profiles of B*0702-_GPGHKARVL-specific CD8 T cells from patients #1017 and #1023 analyzed at two distinct time-points corresponding to weeks 48 and 300 (identified by the arrows in A). <b>C.</b> Cumulative analysis of the functional profile of HIV-specific CD8 T-cell responses on the basis of the expression of IFN-γ, IL-2 and perforin in patients with acute (PHI) infection after one year of ART (-T1Y) and after either treatment interruption (-ATI) or after five years of ART (-T5Y). Matched-paired HIV-specific CD8 T-cell responses are considered for the comparison between T1Y and ATI or between T1Y and T5Y. Although TNF-α was also detected, analysis is restricted to the expression of IFN-γ, IL-2 and perforin for clarity. All possible combinations of IFN-γ, IL-2 and perforin are shown on the <i>x</i> axis, whereas the percentages of the distinct cell subsets within HIV-specific CD8 T cells are shown on the <i>y</i> axis. The pie charts summarize the data, and each slice corresponds to a certain combination of functions. Colors in the pie charts are based on the colored boxes at the bottom of the panel. <b>D.</b> PD-1 expression in B*0702-_GPGHKARVL-specific CD8 T cells in patients #1017 and #1023 measured at the two time-points identified with arrows in A. <b>E.</b> Mean fluorescence intensity (MFI) of PD-1 expression in HIV-specific CD8 T cells measured in patients with acute (PHI) HIV infection after 1 year of ART (-T1Y) and after either treatment interruption (-ATI, left panel) or after five years of ART (-T5Y, right panel). <b>F.</b> Proportion of PD-1⁺2B4⁺CD160⁺ cells in B*0702-_GPGHKARVL-specific CD8 T cells in patients #1017 and #1023 measured at the two time-points identified with arrows in A. <b>G.</b> Proportion of PD-1⁺2B4⁺CD160⁺ cells in HIV-specific CD8 T cells measured in patients with acute (PHI) HIV infection after 1 year of ART (-T1Y) and after either treatment interruption (-ATI, left panel) or after five years of ART (-T5Y, right panel).</p

Functional avidity of HIV-specific CD8 T-cell responses.

<p><b>A.</b> Representative examples of functional avidity of HIV-specific CD8 T-cell responses in patients with acute (PHI-B-09; circles), chronic progressive (CP-B-08; squares) or chronic non-progressive (LTNP-2082; triangles) infection after stimulation with decreasing concentrations of B*0801-_EIYKRWII peptides. The dashed line corresponds to half of the maximal response allowing the extrapolation of the 50% effect concentration (EC₅₀). <b>B.</b> Cumulative analysis of the functional avidity of HIV-specific CD8 T cells during acute (PHI-B), untreated chronic progressive (CP-B) and non-progressive infection (LTNP). Medians and interquartile ranges are shown and each point represents one HIV-specific CD8 T-cell response. <b>C.</b> Cumulative analysis of the frequency of HIV-specific CD8 T cells during acute (PHI-B), untreated chronic progressive (CP-B) and non-progressive infection (LTNP). <b>D.</b> Functional avidity of HIV-specific CD8 T-cell responses against common optimal epitopes recognized by patients with acute (PHI-B), chronic progressive (CP-B) or not progressive (LTNP) infections. The red line (right panel) shows the functional avidity of B*2705-_KRWIILGLNK (KK10)-specific CD8 T-cell response.</p

Associations between functional avidity of HIV-specific CD8 T-cell responses and other immunological parameters.

<p>Associations between the functional avidity of HIV-specific CD8 T-cell responses and the frequency of HIV-specific CD8 T cells co-expressing CD28 and CD27 (<b>A</b>) or PD-1, 2B4 and CD160 (<b>B</b>).</p