Genetic Polymorphism at CCL5 Is Associated With Protection in Chagas' Heart Disease: Antagonistic Participation of CCR1+ and CCR5+ Cells in Chronic Chagasic Cardiomyopathy.

Chronic cardiomyopathy is the main clinical manifestation of Chagas disease (CD), a disease caused by Trypanosoma cruzi infection. A hallmark of chronic chagasic cardiomyopathy (CCC) is a fibrogenic inflammation mainly composed of CD8+ and CD4+ T cells and macrophages. CC-chemokine ligands and receptors have been proposed to drive cell migration toward the heart tissue of CD patients. Single nucleotide polymorphisms (SNPs) in CC-chemokine ligand and receptor genes may determine protein expression. Herein, we evaluated the association of SNPs in the CC-chemokines CCL2 (rs1024611) and CCL5 (rs2107538, rs2280788) and the CCL5/RANTES receptors CCR1 (rs3181077, rs1491961, rs3136672) and CCR5 (rs1799987) with risk and progression toward CCC. We performed a cross-sectional association study of 406 seropositive patients from endemic areas for CD in the State of Pernambuco, Northeast Brazil. The patients were classified as non-cardiopathic (A, n = 110) or cardiopathic (mild, B1, n = 163; severe, C, n = 133). Serum levels of CCL5 and CCL2/MCP-1 were elevated in CD patients but were neither associated with risk/severity of CCC nor with SNP genotypes. After logistic regression analysis with adjustment for the covariates gender and ethnicity, CCL5 -403 (rs2107538) CT heterozygotes (OR = 0.5, P-value = 0.04) and T carriers (OR = 0.5, P-value = 0.01) were associated with protection against CCC. To gain insight into the participation of the CCL5-CCR5/CCR1 axis in CCC, mice were infected with the Colombian T. cruzi strain. Increased CCL5 concentrations were detected in cardiac tissue. In spleen, frequencies of CCR1+ CD8+ T cells and CD14+ macrophages were decreased, while frequencies of CCR5+ cells were increased. Importantly, CCR1+CD14+ macrophages were mainly IL-10+, while CCR5+ cells were mostly TNF+. CCR5-deficient infected mice presented reduced TNF concentrations and injury in heart tissue. Selective blockade of CCR1 (Met-RANTES therapy) in infected Ccr5-/- mice supported a protective role for CCR1 in CCC. Furthermore, parasite antigen stimulation of CD patient blood cells increased the frequency of CCR1+CD8+ T cells and CCL5 production. Collectively, our data support that a genetic variant of CCL5 and CCR1+ cells confer protection against Chagas heart disease, identifying the CCL5-CCR1 axis as a target for immunostimulation.

Increase in the expression of CD4 + CD25+ lymphocytic T cells in the indeterminate clinical form of human Chagas disease after stimulation with recombinant antigens of Trypanosoma cruzi.

PURPOSE: Regulatory T cells are involved in the clinical course of chronic Chagas disease, possibly because they exercise a control in the patient's inflammatory response to Trypanosoma cruzi. This study analyzed the levels of CD4 + CD25+ T cells in chronic Chagas disease patients after in vitro stimulation of the peripheral blood mononuclear cells with CRA (Cytoplasmic Repetitive Antigen) or FRA (Flagellar Repetitive Antigen) T. cruzi antigens. METHODS: Groups of patients with the cardiac form and indeterminate form; and non-infected individuals, were selected. The CD4 + CD25+ T lymphocyte population, as well as the FoxP3 expression and the IL10 production, were evaluated by flow cytometry after stimulation with CRA or FRA. RESULT: The IND group presented higher levels of CD4 + CD25+ T cells than the CARD group. However, there was no evidence of a relationship between FoxP3 and IL10 with any of the chronic forms. CONCLUSIONS: Our results suggest the possible involvement of CD4 + CD25+ T cells specific to CRA and FRA in controlling the progression of clinical outcomes. Though, further studies are needed to define which mechanisms activate regulatory T cells and lead to pathology control in chronic human Chagas disease.

IL-10 and IFN-γ gene expression in chronic Chagas disease patients after in vitro stimulation with recombinant antigens of Trypanosoma cruzi.

Along with several other aspects of Chagas disease, the mechanisms responsible for the different clinical outcomes observed in chronic infected individuals have not yet been clarified. It is believed that the host immune response to the parasite plays an important role in the development of the pathology. Therefore, the aim of this study was to evaluate the relationship between IL-10 and IFN-γ gene expression profile, after in vitro stimulation of peripheral blood mononuclear cells (PBMC) with Trypanosoma cruzi recombinant antigens CRA (cytoplasmatic repetitive antigen) and FRA (flagellar repetitive antigen), and the clinical forms of chronic Chagas disease. Twenty patients with the cardiac form of the disease (CARD), of whom 10 had the mild cardiac form (CARD 1) and 10 the severe cardiac form (CARD 2), and 20 patients with the indeterminate form (IND), were selected at the Chagas Disease Unit of the Oswaldo Cruz University Hospital, University of Pernambuco, Recife, Pernambuco, Brazil. The PBMCs of these individuals were cultured in the presence of CRA or FRA for 3 days and IL-10 and IFN-γ gene expression was evaluated by detection of its messenger RNA using Real Time Quantitative PCR. Although no significant difference was observed between the groups of individuals studied, we found that most patients with IND displayed high levels of IFN-γ gene expression, while the majority of patients with CARD 1 presented high levels of IL-10. The results of this study thus highlight the important role that inflammatory cytokines play in patients with the IND group controlling for parasite replication, and that anti-inflammatory cytokines play in determining susceptibility to progression to symptomatic clinical forms of the disease.