mDC from <i>S. haematobium</i>-infected subjects have an altered MAPK signaling, but not TLR4 expression profile.

<p>(A) TLR4 expression was analysed on mDC present in PBMC following the gating strategy as described in legend of <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0000667#pntd-0000667-g001" target="_blank">Fig. 1</a>. (B and C) 20 and 60 minutes after stimulation with LPS, mDC were intracellularly stained for (B) phospophorylated p38 and ERK and (C) the ratio between <i>p</i>-ERK and <i>p</i>-p38 was determined by dividing the respective mean fluorescence intensities per sample. Each group represents data from 5 donors. (A) Bars represent mean + SD. (B and C) box plots represent 25–75 percentile range with error bars showing minimum to maximum.</p

mDC from <i>S. haematobium</i>-infected subjects have a reduced T cell activating capacity due to lower HLA-DR expression.

<p>LPS matured mDC were cocultured with allogeneic naïve T helper cells for 6 d after which (A) T cell expansion was determined with a counting chamber and (B) Intracellular cytokine production or (C) CD25 and HLA-DR expression was assayed by FACS, 6 h after restimulation with phorbol 12-myristate 13-acetate (PMA) and ionomycin in the presence of brefeldin A for the last 2 h. (D+E) LPS-matured mDC from European controls were cocultured with allogeneic naïve T helper cells for 6 d in the presence of depicted neutralizing antibodies and T cells were counted as in (A). T cell counts are shown as (D) absolute values or (E) relative to the control condition. (A,B,D) Horizontal bars represent mean based on data from 9 donors in each group. (C) Box plots represent 25–75 percentile range with error bars showing minimum to maximum based data of 9 donors in each group. (E) Bars represent mean + SD of 3 independent experiments.</p

Reduced frequencies of mDC and pDC in blood from <i>S. haematobium</i>-infected subjects.

<p>(A) Blood DC were identified in fixed PBMC as HLA-DR⁺/CD14⁻/CD19⁻ cells and subsequently subdivided into mDC and pDC on the basis of positive staining for BDCA-1 and BDCA-2, respectively. Data from one representative donor is shown. (B) Frequencies of blood DC subsets in total PBMC (C) and their surface expression of HLADR, CD80, CD86 and CCR7 was determined by following the gating strategy shown in (A). (B) Box and whiskers with 10–90% percentile are shown. (C) Bars represent mean + SD. (B+C) Each group represents data from 20 donors.</p

Study population.

Δ<p>, based eggs present in 10 ml urine trapped by a 12 µm filter; *, based on Giemsa-stained whole blood smear; ^#, based on presence of microfilaria in cell cultures; ^♦, X² analysis with Fisher's exact test; ND, not determined.</p

mDC, but not pDC, from <i>S. haematobium</i>-infected subjects have impaired TLR responses.

<p>Isolated blood DC were stimulated with 100 ng/ml LPS (mDC), 1 µg/ml R848 (mDC & pDC) or 1 µg/ml CpG (pDC) for 40 h. (A–D) LPS- and R848-matured mDC and CpG and R848 stimulated pDC were analysed for maturation marker expression by FACS. Expression of surface marker is either shown (A+C) as fold increase relative to medium or (B+D) as absolute mean fluorescence intensity. (E) Cytokines levels in 40 h culture supernatants from TLR-stimulated DC were determined by multiplex Luminex. Values represent cytokines concentration from which medium control cytokines levels have been subtracted. (A+C) box plots represent 25–75 percentile range with error bars showing minimum to maximum. *, p<0,05; **, p<0,01; ***, p<0,001 for significant differences in fold increase in maturation marker expression relative to the medium-stimulated DC. (B, D and E) Bars represent mean + SD. Each group represents data from 20 donors, except for (E) in which R848-stimulated pDC cytokine data are represented by 4 and 5 donors for the infected and un-infected group, respectively. ND: not detectable.</p

Multivariable logistic regression analysis for delivery of an infant with low birth weight.

<p>LBW = low birth weight, OR = Odds Ratio, IPTp = intermittent preventive treatment of malaria during pregnancy.</p

Univariable analysis of adolescent compared to adult women.

<p>LBW = low birth weight, IPTp = intermittent preventive treatment of malaria during pregnancy.</p

Birth weight of term-newborns born to adolescent and adult mothers for each week of gestation.

<p>Bars show mean birth weight and standard deviations for adolescent (≤16 years) and adult (>16 years) mothers.</p

Differences in Innate Cytokine Responses between European and African Children

<div><p>Although differences in immunological responses between populations have been found in terms of vaccine efficacy, immune responses to infections and prevalence of chronic inflammatory diseases, the mechanisms responsible for these differences are not well understood. Therefore, innate cytokine responses mediated by various classes of pattern-recognition receptors including Toll-like receptors (TLR), C-type lectin receptors (CLRs) and nucleotide-binding oligomerisation domain-like receptors (NLRs) were compared between Dutch (European), semi-urban and rural Gabonese (African) children. Whole blood was stimulated for 24 hours and the pro-inflammatory tumor necrosis factor (TNF) and the anti-inflammatory/regulatory interleukin-10 (IL-10) cytokines in culture supernatant were measured by enzyme-linked immunosorbent assay (ELISA). Gabonese children had a lower pro-inflammatory response to poly(I:C) (TLR3 ligand), but a higher pro-inflammatory response to FSL-1 (TLR2/6 ligand), Pam3 (TLR2/1 ligand) and LPS (TLR4 ligand) compared to Dutch children. Anti-inflammatory responses to Pam3 were also higher in Gabonese children. Non-TLR ligands did not induce substantial cytokine production on their own. Interaction between various TLR and non-TLR receptors was further assessed, but no differences were found between the three populations. In conclusion, using a field applicable assay, significant differences were observed in cytokine responses between European and African children to TLR ligands, but not to non-TLR ligands.</p></div

Whole blood cytokine responses to TLR ligands.

<p>A) TNF responses to poly(I:C), FLS-1, Pam3 and LPS in European children (the Netherlands), and semi-urban and rural African children (Gabon). B) IL-10 responses to TLR stimulation. C) Pro/anti-inflammatory ratio as calculated by TNF/IL-10 ratio. *p<0.05, **p<0.01, ***p<0.001.</p