Interethnic and intraethnic differences in the TNF-α/IL-10 and IFN-γ/IL-10 ratios upon TLR stimulation of PBMCs.

<p>PBMCs (1 million/ml) were stimulated or not with LPS (100 ng/ml), CpG-A (3 µg/ml) or imiquimod (1 µg/ml). After 16 hours, supernatants were collected, cytokine levels were analyzed and the TNF-α/IL-10 (Figure 4A) and IFN-γ/IL-10 (Figure 4B) ratios were calculated. The samples from 77 children were categorized as uninfected Dogon (n = 20), infected Dogon (n = 20), uninfected Fulani (n = 23) and infected Fulani (n = 14). The TNF-α/IL-10 ratio (Figure 4A) was significantly lower in the infected Dogon as compared to their uninfected peers when stimulated with CpG or imiquimod. This resulted in significantly higher TNF-α/IL-10 ratio of infected Fulani as compared to the infected Dogon. Due to the severe suppression of IFN-γ the IFN-γ/IL-10 ratios (Figure 4B) were severely suppressed in the LPS and CpG stimulated cells of the Dogon and therefore higher in the infected Fulani than in the infected Dogon. The boxplots illustrate the medians and the 25^th and 75^th quartile and the whiskers represent the 10% and 90% percentiles. Data were analyzed using Mann-Whitney rank sum test. *; p≤0.05. **; p<0.01. ***; p<0.001.</p

TLR-induced cytokine responses in PBMCs from Dogon and Fulani children with or without <i>P. falciparum</i> infection.

<p>PBMCs (1 million/ml) were stimulated or not with LPS (100 ng/ml), CpG-A (3 µg/ml) or imiquimod (1 µg/ml). After 16 hours, supernatants were collected and analysed for the presence of cytokines. The samples were categorized as uninfected Dogon (n = 20), infected Dogon (n = 20), uninfected Fulani (n = 14) and infected Fulani (n = 23). The levels of IL-1β, IL-6, IL-10, TNF-α and INF-γ were severely supressed in the infected Dogon as compared to their uninfected peers when the cells had been stimulated with either LPS or CpGA. As a result, the levels of the infected Fulani were higher than infected Dogon for all cytokines except for IFN-α when stimulated with LPS and except for IL-1β and IL-10 when stimulated with CpG. Similarly, when stimulated with imiquimod cells of the infected Fulani secreted higher levels of IL-6, IFN-α, TNF-α and INF-γ than the infected Dogon due to the suppressed secretion of Dogon when undergoing infection. Data presented as box plots illustrate the medians and the 25^th and 75^th quartile and the whiskers represent the 10% and 90% percentiles. Data were analyzed using Mann-Whitney rank sum test. *; p≤0.05. **; p<0.01. ***; p<0.001.</p

Interethnic and intraethnic differences in the frequency and activation markers of blood DCs.

<p>PBMCs from the study participants were stained with monoclonal antibodies for subsequent flow cytometric analysis using a FACSCalibur. The samples were categorised as uninfected Dogon (n = 20), infected Dogon (n = 20), uninfected Fulani (n = 13) and infected Fulani (n = 10). An increase was observed of BDCA2⁺ and BDCA3⁺ cells in the circulation of infected Dogon whereas the opposite was seen for the Fulani (Figure 1A). The expression of activation marker HLA-DR (Figure 1B) and CD86 (Figure 1C) was lower in BDCA2⁺ and BDCA3⁺ cells of the Dogon when undergoing infection while it was increased in the Fulani. It is known that DCs travel to the lymph nodes upon activation. Therefore, it is possible that the lower numbers seen in the infected Fulani is a consequence of activation. The boxplots illustrate the medians and the 25^th and 75^th quartile and the whiskers represent the 10% and 90% percentiles. Data were analyzed by Mann-Whitney rank sum test. *; p≤0.05. **;p<0.01. ***; p<0.001.</p

The log₁₀ p-values from applying SNP association tests for malaria and clinical phenotypes (from a logistic regression adjusted for age and season).

<p>(<b>a</b>) Clinical malaria.* dashed line represents a p-value of 0.003, polymorphisms are ordered by chromosome (coloured differently) and position (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075675#pone.0075675.s001" target="_blank">Tables S1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075675#pone.0075675.s002" target="_blank">S2</a>).(<b>b</b>) Asymptomatic malaria.* dashed line represents a p-value of 0.003, polymorphisms are ordered by chromosome (coloured differently) and position (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075675#pone.0075675.s001" target="_blank">Tables S1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075675#pone.0075675.s002" target="_blank">S2</a>). (<b>c</b>) Any malaria. * dashed line represents a p-value of 0.003, polymorphisms are ordered by chromosome (coloured differently) and position (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075675#pone.0075675.s001" target="_blank">Tables S1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075675#pone.0075675.s002" target="_blank">S2</a>). (<b>d</b>) Spleen enlargement. * dashed line represents a p-value of 0.003, polymorphisms are ordered by chromosome (coloured differently) and position (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075675#pone.0075675.s001" target="_blank">Tables S1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075675#pone.0075675.s002" target="_blank">S2</a>). (<b>a</b>) Parasite positivity. * dashed line represents a p-value of 0.003, polymorphisms are ordered by chromosome (coloured differently) and position (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075675#pone.0075675.s001" target="_blank">Tables S1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075675#pone.0075675.s002" target="_blank">S2</a>).</p

The log₁₀ p-values from applying SNP association tests for immunological titre phenotypes (from a linear regression adjusted for age and season).

<p>(<b>a</b>) CSP. * dashed line represents a p-value of 0.003, polymorphisms are ordered by chromosome (coloured differently) and position (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075675#pone.0075675.s001" target="_blank">Tables S1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075675#pone.0075675.s002" target="_blank">S2</a>). (<b>b</b>) AMA1. * dashed line represents a p-value of 0.003, polymorphisms are ordered by chromosome (coloured differently) and position (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075675#pone.0075675.s001" target="_blank">Tables S1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075675#pone.0075675.s002" target="_blank">S2</a>). (<b>c</b>) MSP1. * dashed line represents a p-value of 0.003, polymorphisms are ordered by chromosome (coloured differently) and position (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075675#pone.0075675.s001" target="_blank">Tables S1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075675#pone.0075675.s002" target="_blank">S2</a>). (<b>d</b>) MSP2. * dashed line represents a p-value of 0.003, polymorphisms are ordered by chromosome (coloured differently) and position (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075675#pone.0075675.s001" target="_blank">Tables S1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075675#pone.0075675.s002" target="_blank">S2</a>). (<b>e</b>) <b>Total IgE</b>. * dashed line represents a p-value of 0.003, polymorphisms are ordered by chromosome (coloured differently) and position (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075675#pone.0075675.s001" target="_blank">Tables S1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075675#pone.0075675.s002" target="_blank">S2</a>).</p

In-vitro cytokine measurements after PBMC stimulation with <i>Yersinia enterocolitica</i>, <i>Yersinia pseudotuberculosis</i> and <i>Yersinia pestis</i>.

<p>Caucasian individuals bearing a normal (NOD2 wt, N = 10) or homozygous for the 3020insC NOD2 mutation (NOD2 del, N = 5) were stimulated with the various <i>Yersinia spp</i>. TNF (A) and IL-1β (B) were measured by ELISA after 24-hour stimulation. Values represent mean + SD for each group of volunteers. P-values for differences between caspase-12 genotype were calculated with the Mann-Whitney test. *P<0.05, **P≤0.01.</p

Caspase-12 genotypes and allele frequency in two cohorts of Mali volunteers, one collected in 2006 and one in 2007.

a<p>Population do not violate Hardy-Weinberg equilibrium.</p>b<p>Distribution of Caspase-12 genotypes between 2006 and 2007 populations did not differ (χ²-test).</p

In-vitro cytokine measurements after whole blood LPS and Pam3Cys stimulation between the caspase-12 genotypes from the volunteers enrolled in 2006.

<p>Stimulation was performed with 1, 10 and 100 ng/ml LPS (<i>E. coli</i>) and 10 µg/ml Pam3Cys. TNF (A), IL-1β (B) and IL-10 (C) were measured by ELISA in the supernatant after 24 hour stimulation. Volunteers were grouped as homozygous bearing the nonfunctional caspase-12 genotype (S/S), heterozygous bearing the nonfunctional and functional caspase-12 (S/L), and homozygous bearing the functional caspase-12 (L/L). Numbers of volunteers included for each cytokine are S/S = 33, S/L = 16 and L/L = 1. Values represent mean + SD for each group of volunteers. P-values for differences between caspase-12 genotype were calculated with the Mann-Whitney test. *P<0.05.</p

In-vitro cytokine measurements after whole blood LPS and Pam3Cys stimulation between the caspase-12 genotypes from the volunteers enrolled in 2007.

<p>Stimulation with LPS was performed with 10 ng/ml. TNF (A), IL-1β (B) and IL-10 (C) were measured by ELISA after 24-hour stimulation. Volunteers were grouped as homozygous bearing the nonfunctional caspase-12 genotype (S/S), heterozygous bearing the nonfunctional and functional caspase-12 (S/L) and homozygous that bear the functional caspase-12 (L/L). Numbers of volunteers included for each cytokine are S/S = 24, S/L = 22 and L/L = 1. Values represent mean + SD for each group of volunteers. P-values for differences between caspase-12 genotype were calculated with the Mann-Whitney test.</p