Project description:Extracellular vesicles (EVs) from parasite protozoa can induce efficient immunity, which makes them useful for probable next-generation vaccines, more specifically against Trypanosoma cruzi infection. Howevver, the mechanisms involved in the protective responses have not been fully elucidated. To investigate the potential use of trypanosomatid EVs as immunogens against Chagas disease we tested the effect of EVs stimulation from Trypanosoma rangeli and Phytomonas serpens compared to T. cruzi response in vitro and in vivo, besides performing a proteomics evaluation of the EVs.Overall, our results pointed out that EVs of T. cruzi-correlated parasites have antigenic potential. Despite minor effects in the modulation of antigen presentation cells surface receptors and in in vitro T. cruzi infection, T. rangeli and P. serpens EVs can contribute to reducing T. cruzi infectivity in Balb/c mice.
Project description:An efficient innate immune recognition of the intracellular parasite T. cruzi is crucial for host protection against development of Chagas disease, which often leads to multiple organ damage, particularly the heart leading to cardiomyopathy. Mechanisms modulated by MyD88 have been shown to be necessary for resistance against T, cruzi infection. Recently, Nod-like receptors have been shown to play an important role as innate immune sensors, particularly as they relate to inflammasome function, caspase activation, and inflammatory cytokine production. In this study, we aimed to investigate the participation of innate immune responses in general, and inflammasomes in particular, in heart inflammation and cardiac damage upon infection with the T. cruzi parasite. We used microarrays to gain insight into gene expression in the cardiac tissue of mice infected with the causative agent of Trypanosoma cruzi, and identified distinct classes of up-regulated genes during this process, including important genes involved in inflammasome activation and innate immune responses in general. The hearts of C57BL/6 mice day 18 post-infection with a Y strain of the parasite T. cruzi, and uninfected controls were extraced for RNA extraction and hybridization on Affymetrix microarrays. We sought to compare gene expression among two groups of mice, and so extracted the hearts of 3 control uninfected mice, and of 3 infected mice 18 days post-infection.
Project description:Investigate transcriptional responses of T. cruzi infected hiPSC-derived cardiomyocytes from chagasic patients with and without cardiomyopathy.
Project description:Mice were infected with Trypanosoma cruzi strains CL, X10/4 or X10/7. Hearts were collected 12 days, 3 months and 5 months post-infection from infected mice and uninfected age-matched controlled. Hearts were divided into four sections from the base of the heart to the apex. Metabolites were extracted by a two-step protocol, first with 50% methanol (aqueous extract) then with 3:1 dichloromethane:methanol (organic extract). Liquid chromatography separation was performed on a Kinetex C8 LC Column 50 x 2.1 mm, with water+0.1% formic acid and acetonitrile+0.1% formic acid as mobile phases.
Project description:Mice were infected with Trypanosoma cruzi parasites, followed by treatment with benznidazole. Controls include uninfected mice and mice treated with isoproterenol to induce cardiac failure through chemical mechanisms. Urine was collected pre- and post-treatment and extracted using methanol, followed by LC separation using polar C18 LC and positive mode MS/MS data acquisition.
Project description:Fecal samples were collected from Trypanosoma cruzi-infected (strain CL+Luc) and uninfected mice for up to 3 months post-infection. Samples were extracted with 50% methanol.
Project description:These assays represent an antigen discovery screening, and epitope mapping characterization. In this screening two complete proteomes from Trypanosoma cruzi, from two different strains (CL-Brener, Sylvio X10), were displayed in the form of short peptides (tiling array, overlapped) and assayed with pooled serum samples (antibodies) from Chagas Disease patients and matched negative (healthy) subjects selected from 6 geographic regions across the Americas. Peptide arrays (slides) were incubated with pooled serum samples (primary antibodies), washed, and then incubated with a fluorescently-labeled anti-human IgG commercial antibody (secondary antibodies). Raw readouts of fluoresence (signal), as well as normalized signal values are provided in this submission for all samples analyzed. All samples were analyzed in duplicate.