Patient characteristics showing differences between HIV infected and uninfected participants.

<p>Patient characteristics showing differences between HIV infected and uninfected participants.</p

Influence of HIV infection on dendritic cell migration and activation in ectocervical explants.

<p>DC migration and activation was compared between HIV-infected (n = 6) and uninfected participants (n = 8). Ectocervical explants from HIV-infected and uninfected participants were stimulated for 24 hours with Medium, cytokines or TLR agonists. Migration and activation of DCs was evaluated and compared between the two groups of participants for the three DC subsets and stimulation conditions. Data presented as Box and Whisker plots of fold migration above background (medium alone). Mann-Whitney test was used for statistical analyses. * P-values <0.05</p

Dendritic cell migration and activation from ectocervical and endocervical tissue explants.

<p>Cervical explants were exposed to cytokines (TNF-α, IL-1β, IL-8 and MIP-1β) or TLR agonists (LPS, R848, or PAM3) for 24 hours. Frequencies of DCs that had migrated out of the tissue blocks into collected culture medium, and their activation status was measured in tissue from (A) the ectocervix, and (B) the endocervix. Migration index indicates the ratio of stimulated to unstimulated DC frequencies. Activation index indicates the ratio of stimulated to unstimulated DC HLA-DR MFI. Data is presented as median with interquartile range fold induction or migration above background (medium). Each experiment was performed once and different samples were processed and stimulated for the different conditions: Medium (n = 14), cytokines (n = 14), LPS (n = 14), PAM3 (n = 14) and R848 (n = 14) for ectocervical explants; and Medium (n = 17), cytokines (n = 16), LPS (n = 17), PAM3 (n = 16) and R848 (n = 16) for endocervical explants. Wilcoxon matched-pairs signed rank test was used for statistical analyses. * P-values <0.05.</p

Identification of cervical DC subsets from (ecto) cervical explant cultures.

<p>(A) Plots showing the exclusion of dead cells (VIVID⁺), doublets, neutrophils/epithelial cells (CD66a/c/e⁺) and monocytes (CD66a/c/e⁻ CD14⁺). (B) Dendritic cell subsets were identified as mDCs (CD66a/c/e⁻ CD14⁻ CD11c⁺); pDCs (CD66a/c/e⁻ CD14⁻ CD11c⁻ CD123⁺); and LCs (CD66a/c/e⁻ CD14⁻ CD11c⁻ CD123⁻ CD1a⁺); and frequencies of cervical tissue migrating DCs determined, following different stimulation conditions. (C) Histogram showing up-regulation of HLA-DR expression by cervical explant migrating mDCs. Activation of pDCs and LCs was determined in a similar way as mDCs and Median Fluorescent Intensities (MFI) calculated.</p

Comparison of cytokine concentrations and detection frequencies in genital fluid isolated from menstrual cups and CVLs in a randomised study.

<p>^aIQR: Interquartile range.</p><p>^bLLOD: Lower limit of detection.</p><p>^cMann-Whitney U tests were conducted to compare non-parametric unpaired data.</p><p>^dFisher’s exact tests were conducted to compare detection frequencies.</p><p>§Significant P-value after FDR multiple comparison adjustment.</p><p>Comparison of cytokine concentrations and detection frequencies in genital fluid isolated from menstrual cups and CVLs in a randomised study.</p

Characteristics of participants overall, and stratified by study arm.

<p>Not all the study participants were tested for an STI or BV</p><p>Characteristics of participants overall, and stratified by study arm.</p

Comparison of total antibody (ng ml-1) in MCs (n = 19)- 3A, randomized CVL (n = 20) and matched post-MC CVLs (n = 18) [Log₁₀ MFI*dilution factor ng ml^-1 (MFI/total Ig)]- Correlation plots between total antibody in MCs and matched post MC CVLs- 3B.

<p>Correlation plots (Fig 3C–3F) of HIV specific activity in MCs (n = 18) and matched post-MC CVLs (n = 18) [Log₁₀ MFI*dilution factor ng ml^-1 (MFI/total Ig)] for gag p24- 3C, p66- 3D, gp41- 3E and gp120- 3F.</p

Comparison of median cytokine concentrations observed in genital fluid isolated from CVL, MC and matching subsequent CVL specimens in a randomised study.

<p>Cytokine concentrations were compared in genital fluid isolated from n = 19 MC- 4A and n = 20 randomized CVL, and n = 18 matching MC and subsequently-sampled CVL specimens- 4B. The concentrations of 48 cytokines [(regulatory ○; growth factor ✚; haematopoietic ✖; adaptive ◆; inflammatory ●)] were determined in each specimen by luminex technology and the median of each cytokine concentration was compared by Spearman rank correlation. Data are depicted on Log₁₀ scales. P-values <0.05 were considered significant.</p

Schema of study design.

<p>Schema of study design.</p

HIV specific activity [Log₁₀ MFI*dilution factor ng ml^-1 (MFI/total Ig)] for gag p24- 2A, p66- 2B, gp41- 2C and gp120- 2D in MC (n = 19) and randomized CVL samples (n = 20).

<p>Limit of detection is shown as the dotted line on the figures.</p