Genotype and allele frequencies for the <i>CXCL10 rs3921</i> polymorphism in patients with Chagas disease.

<p>CCC patients were further stratified by left ventricular ejection fraction values.</p><p>Data are no. (%) of patients. Moderate CCC has LVEF>40%. Severe CCC has LVEF≤40%. CI, confidence interval. OR, odds ratio.</p

Chemokine and chemokine receptor expression in myocardium from patients bearing the <i>CXCL9</i> and <i>CXCL10</i> SNPs.

<p>Expression of chemokines and chemokine receptors in myocardium samples from patients bearing the <i>CXCL9</i> rs10336 SNP (A). Expression of chemokines and chemokine receptors in myocardium samples from patients bearing <i>CXCL10</i> rs3921 SNP (B). Real-time qPCR analysis of mRNA levels expression in myocardium samples from patients bearing the <i>CXCL9</i> rs10336 <i>CC</i> (n = 5) or <i>CT</i> (n = 9) genotypes (A) and <i>CXCL10</i> rs3921 <i>GG</i> (n = 5) or <i>CC</i> or <i>CG</i> (n = 9). The bar stands for the median.</p

Genotype and allele frequencies for the <i>CXCL9</i> rs10336 polymorphism in patients with Chagas disease.

<p>CCC patients were further stratified by left ventricular ejection fraction values.</p><p>Data are no. (%) of patients. Moderate CCC has LVEF>40%. Severe CCC has LVEF≤40%. CI, confidence interval. OR, odds ratio.</p

Correlation between histopathologically assessed intensity of myocarditis and mRNA expression of CXCL9 in CCC myocardium.

<p>Two-tailed p value obtained with Spearman's nonparametric correlation. n = 14.</p

Histopathological analysis and localization of CCR5⁺, CXCR3⁺, CCR4⁺, CCL5⁺ and CXCL9⁺ cells in heart tissue.

<p>Severe myocarditis with intense mononuclear inflammatory infiltrate in the heart tissues of CCC patient, and absence of myocarditis in the heart tissues of NIC patients and individuals without cardiomyopathies (N). In (A), imagens representative of heart fragments from CCC, NIC and N individuals. Haematoxylin-eosin stain. Representative images of immunofluorescence confocal microscopy from heart tissue sections from CCC and NIC (B). Sections were stained with primary antibodies against CCR5, CXCR3, CCR4, CCL5 and CXCL9 stained with AF633-conjugated anti-mouse IgG (red) and counterstained with DAPI (blue), as described in <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0001867#s2" target="_blank">Methods</a>. CTR- IgG isotype controls.</p

Genotype and allele frequencies for the <i>CCR5</i> rs1799988 polymorphism in patients with Chagas disease.

<p>CCC patients were further stratified by left ventricular ejection fraction values.</p><p>Data are no. (%) of patients. Moderate CCC has LVEF>40%. Severe CCC has LVEF≤40%. CI, confidence interval. OR, odds ratio.</p

Characteristics of patients and control donors whose myocardial samples were used in gene expression studies.

<p>Age (years). M (male). F (female). CCC (Chronic Chagasic Cardiomyopathy). NIC (Non-inflammatory Cardiomyopathy). IDC (Idipathic Dilated Cardiomyopathy). IC (Ischemic Cardiomyopathy). Normal heart donors were subject to ventilator and vasoactive drugs, and had been under life support for an average of 48 hours. Characterization of the samples as myocarditis, fibrosis and hypertrophy; reference values for the presence of myocarditis and fibrosis: 0: absent; 1+: slight; 2+: moderate; 3+: intense; hypertrophy: Y: yes N: no. EF (Ejection Fraction) was always less than 40%. LVDD (left ventricle diastolic diameter) normal reference value: diameter 39–55 mm. Values of mRNA expression (RT-qPCR) of the atrial natriuretic peptide (ANP) and the brain natriuretic peptide (BNP). -, Control samples were used as calibrators in the RT-qPCR reaction and have no relative expression value using the 2^−ΔΔCt calculation method as described in <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0001867#s2" target="_blank">Methods</a>.</p

Expression of chemokines and chemokine receptors in myocardium.

<p>Expression of chemokine receptors (A) in CCC and NIC myocardium. Expression of chemokines (B) in CCC and NIC myocardium. Real-time qPCR analysis of mRNA levels expression in CCC and NIC myocardium. After normalization to GAPDH mRNA, the relative expression was plotted in comparison to N group and data were calculated with the 2^−ΔΔCt method, as described in <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0001867#s2" target="_blank">Methods</a>. The horizontal bar stands for the median. Dotted lines indicate two-fold increase or decrease expression as compared with the control group.</p

Correlation analysis for the rs640249 polymorphism in two different reference populations.

<p>Genotype data from the European reference population (CEU) and from the West Africa reference population (YRI) were downloaded from HapMap. A 2 Mb region surrounding the ACTC1 gene was analyzed in these two populations. The data were analyzed with Haploview Software. <b>A)</b> Correlations were assessed by calculating r² values. The rs640249 polymorphism was found to be correlated with three other polymorphisms in the CEU reference population (rs641563; rs639735; rs479623). <b>B</b>) In the West Africa reference population, the rs640249 polymorphism was correlated only with rs641563. </p

The rs640249A-rs641563A haplotype is associated with resistance to CCC.

<p>Based on the genotypes obtained, we performed a haplotype analysis of the rs7719175 and rs1800925 polymorphisms. Only three haplotypes were found in our study population: <b>A</b>) Distribution of the three main haplotype combinations in the CCC patients and ASY subjects: homozygous rs640249C-rs641563C (black bar); heterozygous rs640249C-rs641563C + rs640249A-rs641563A (gray bar) and homozygous rs640249A-rs641563A (white bar). <b>B</b>) Haplotype combinations between cases and controls, taking sex into account. <b>C</b>) Distribution of the three main haplotype combinations between patients with severe and moderate CCC, taking sex into account.</p