Background Non-human primates have been shown to be useful models for Chagas disease. We previously reported that natural T. cruzi infection of cynomolgus macaques triggers clinical features and immunophenotypic changes of peripheral blood leukocytes resembling those observed in human Chagas disease. In the present study, we further characterize the cytokine-mediated microenvironment to provide supportive evidence of the utility of cynomolgus macaques as a model for drug development for human Chagas disease. Methods and findings In this cross-sectional study design, flow cytometry and systems biology approaches were used to characterize the ex vivo and in vitro T. cruzi-specific functional cytokine signature of circulating leukocytes from TcI-T. cruzi naturally infected cynomolgus macaques (CH). Results showed that CH presented an overall CD4+-derived IFN-γ pattern regulated by IL-10-derived from CD4+ T-cells and B-cells, contrasting with the baseline profile observed in non-infected hosts (NI). Homologous TcI-T. cruzi-antigen recall in vitro induced a broad pro-inflammatory cytokine response in CH, mediated by TNF from innate/adaptive cells, counterbalanced by monocyte/B-cell-derived IL-10. TcIV-antigen triggered a more selective cytokine signature mediated by NK and T-cell-derived IFN-γ with modest regulation by IL-10 from T-cells. While NI presented a cytokine network comprised of small number of neighborhood connections, CH displayed a complex cross-talk amongst network elements. Noteworthy, was the ability of TcI-antigen to drive a complex global pro-inflammatory network mediated by TNF and IFN-γ from NK-cells, CD4+ and CD8+ T-cells, regulated by IL-10+CD8+ T-cells, in contrast to the TcIV-antigens that trigger a modest network, with moderate connecting edges. Conclusions Altogether, our findings demonstrated that CH present a pro-inflammatory/regulatory cytokine signature similar to that observed in human Chagas disease. These data bring additional insights that further validate these non-human primates as experimental models for Chagas disease.
Author summary Trypanosoma cruzi is the causative agent of Chagas disease; millions of people are infected with this parasite. One of the major challenges to manage infected patients is the low efficacy of currently available treatments, especially during chronic infection. Different T. cruzi genotypes are known to differ in response to existing drugs (e.g., TcI is quite resistant), and differences among individuals in immune response also are believed to play a role determining therapeutic efficacy. Experimental models and in vitro systems have been proposed for rational searches for new compounds for treating infected individuals, optimally before the infection becomes clinically manifested as Chagas disease. In the field of drug development, the non-human primate models offer a unique and valuable contribution, as a consequence of patho-physiological similarities that mimic many human diseases, including Chagas disease. In the present study, we further investigated the functional features of the immune response triggered by TcI T. cruzi-infection, characterizing the ex vivo as well as the in vitro cytokine microenvironment, upon T. cruzi-antigen recall. Our results revealed that chronically infected cynomolgus macaques display a similar ex vivo cytokine signature to that observed in chronic human Chagas disease. Moreover, CH macaques display a complex cross-talk among the cytokine+ leukocyte subsets, enhanced by TcI T. cruzi-antigen recall in vitro. These findings provide additional insights that further validate these non-human primates as experimental models for rational development of new therapeutic agents for Chagas disease.