Comparison of specific phenotype frequencies based on CD11c expression in blood samples from healthy women.

<p>Comparison of the frequency of CD8, γδTCR, MAIT and iNKT (Vα24) in CD11c⁺ and CD11c^-, CCR7^- CD3⁺ T cells from the same individual. (<b>a</b>) Fresh blood (n = 10) and (<b>b</b>) PBMC (n = 6). Gating strategy consisted on the following consecutive gates: lymphocytes, singlets, live CD14^- CD19^- CD3⁺ T cells and CCR7^- CD11c⁺or CD11c^- T cells (<i>see</i> <b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0154253#pone.0154253.s003" target="_blank">S3 Fig</a></b> <i>for further details</i>). Data were analyzed using the paired Student’s t-test.</p

CD11c expression in circulating T cells from healthy young women.

<p>CD11c⁺ cells were analyzed by flow cytometry in the CD4⁺ CCR7⁺/^- and CD4^- CCR7⁺/^- CD3⁺ T cell subsets. Gating strategy consisted on the following consecutive gates: lymphocytes, singlets, live CD3⁺ T cells, CD4⁺ or CD4^- T cells, CCR7⁺ or CCR7^- and lastly CD11c⁺. Each bar represents the mean ± SD of normal donors (n = 13). Data were analyzed by Kruskall-Wallis test with Bonferroni post-test correction.</p

Example of up-regulated genes in activated effector CD11c⁺ vs. CD11c^- T cells in naive mice.

<p>Example of up-regulated genes in activated effector CD11c⁺ vs. CD11c^- T cells in naive mice.</p

Comparison of adhesion molecule frequencies based on CD11c expression in CD4^- T_EM cells from healthy women.

<p>Comparison of the frequency of (<b>a</b>) CCR2, (<b>b</b>) CCR5, (<b>c</b>) CCR9, (<b>d</b>) CCR10, (<b>e</b>) CXCR6, (<b>f</b>) α1β1, (<b>g</b>) α4β7, (<b>h</b>) HLA-DR⁺ CD38⁺ in CD11c⁺ and CD11c^- CD4^- CCR7^- T cells from the same individual (n = 13). Gating strategy consisted on the following consecutive gates: lymphocytes, singlets, live CD3⁺ T cells, CCR7^- CD11c⁺or CD11c^- T cells, CD4^- T cells and expression of the different molecules addressed. Data were analyzed using Wilcoxon matched-paired signed-ranked test.</p

Unconventional phenotypes included in the CD11c⁺ T cell fraction after vaginal Chlamydia infection in mice.

<p>The frequency of the different subsets obtained from combining CD1d-tetramer, CD8α, γδTCR and NK1.1 expression is displayed for each group as a pie chart and as a complementary bar graph. Gating strategy was performed as described in <b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0154253#pone.0154253.g001" target="_blank">Fig 1</a></b> and in Materials and Methods. The frequency of CD11c positive cells in the T cell fraction as the mean ± SD is shown as a white number in the center of the pie chart for each group in blood (top) and genital tract (GT, bottom). Each colored portion of a pie chart indicates the percentage of a specific subset detailed in the bar chart below. The arcs around the pie show the molecule or combination of molecules to which those proportions correspond (see color legend indicating CD1d-tetramer, CD8α, γδTCR and NK1.1). *Indicates p<0.05 by Student’s t test analyses only for values >5% of the total CD11c⁺ T cells in vaginally (VAG)-infected mice seven days after infection (n = 4) compared to control (n = 3) animals.</p

Adhesion molecules included in the CD11c⁺ T cell fraction after vaginal Chlamydia infection in mice.

<p>The frequency of the different subsets obtained from combining CCR10, CD8α, CD103 and NK1.1 expression is displayed for each group as a pie chart and as a complementary bar graph. Gating strategy was performed as described in <b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0154253#pone.0154253.g001" target="_blank">Fig 1</a></b> and in Materials and Methods. The frequency of CD11c positive cells in the T cell fraction as the mean ± SD is shown as a white number in the center of the pie chart for each group in blood (top) and genital tract (GT, bottom). Each colored portion of a pie chart indicates the percentage of a specific subset detailed in the bar chart below. The arcs around the pie show the molecule or combination of molecules to which those proportions correspond (see color legend indicating CCR10, CD8α, CD103 and NK1.1). *Indicates p<0.05 by Student’s t test analyses only for values >5% of the total CD11c⁺ T cells in vaginally (VAG)-infected mice seven days after infection (n = 4) compared to control (n = 3) animals.</p

IFN-γ-secreting γδ T cells in PBMC from healthy women after HMBPP activation.

<p>(<b>a</b>) Comparison of the frequency of γδTCR⁺ CD11c⁺/^- and γδTCR^- CD11c⁺/^- T cells that secrete IFNγ in PBMC from the same individual (n = 6) after 20 hours of (E)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate (HMBPP) activation. Gating strategy consisted on the following consecutive gates: lymphocytes, singlets, CD3⁺ T cells, γδTCR⁺/^-, CD11c⁺/^- and IFNγ/CD69⁺ expression. Each bar represents the mean ± SD of control (squares) and HMBPP-stimulated PBMC (circles) samples. Data were analyzed using the non-parametric Friedman test for repeated measures, with Dunn’s multiple comparisons post-hoc test. Graphs below show the comparison in the frequency of (<b>b</b>) IFN-γ-secreting γδ T cells and (<b>c</b>) CD69 expression in γδ T cells based on CD11c expression in HMBPP-activated PBMC from the same individual (n = 6). Data were analyzed using Wilcoxon matched-paired signed-ranked test.</p

Phenotype of cervically derived T cells from healthy women and analysis by CD11c expression.

<p>(<b>a</b>) Representative dot plots from the T cell subsets extracted from the endocervix of healthy women. Top row shows the consecutive general gating strategy to select CD3⁺ T cells. Bottom row shows different subsets analyzed in the total CD3⁺ T cells and in the CD11c⁺ and CD11c^- T cell fractions. (<b>b</b>) Frequency of CD8, iNKT (Vα24), MAIT, and γδT cells by CD11c expression in T cells obtained from cervical tissue. Each bar represents the mean ± SD of the ectocervix and endocervix of each donor (n = 5). Data were analyzed using Wilcoxon matched-paired signed-ranked test.</p

T cell phenotypes in blood and genital tract of mice by CD11c expression.

<p>Comparison on the frequency of (<b>a</b>) NK1.1, (<b>b</b>) CD8α, (<b>c</b>) CD1d-tetramer, (<b>d</b>) γδTCR, (<b>e</b>) CD103 and (<b>f</b>) CCR10 in CD11c⁺ (squares) and CD11c^- (circles) T cells from the same individual. Gating strategy consisted on a lymphocyte gate based on FSC vs. SSC, followed by doublet exclusion and a CD3⁺ T cells gate. After gating on CD11c⁺ or CD11c^- T cells, surface expression of the different markers was quantified (<i>see</i> <b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0154253#pone.0154253.s002" target="_blank">S2 Fig</a></b> <i>for further details</i>). Each bar represents the mean ± SD of control (white; n = 3 or n = 6) or vaginally (VAG)-infected mice (black; n = 4 or n = 8) seven days after infection. Data were analyzed using the paired Student’s t-test.</p

Prevalence curve of HR-HPV infection by age group.

<p>Prevalence curve of HR-HPV infection by age group.</p