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Immunopeptidomics on T. cruzi – infected MC57G murine fibroblasts

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Mar 27, 2025 version files 4.97 GB

Abstract

Trypanosoma cruzi is a protozoan parasite that causes Chagas disease. Globally 6 to 7 million people are infected by this parasite of which 20-30% will progress to develop Chronic Chagasic Cardiomyopathy (CCC). Despite its high disease burden, no clinically approved vaccine exists for the prevention or treatment of CCC. Developing vaccines that can stimulate T. cruzi-specific CD8+ cytotoxic T cells and eliminate infected cells requires targeting parasitic antigens presented on major histocompatibility complex class I (MHC-I) molecules. We utilized mass spectrometry-based immunopeptidomics to investigate which parasitic peptides are displayed on MHC-I of T. cruzi infected cells. Murine MC57G fibroblasts were infected with T. cruzi trypomastigotes strain Tulahuen. After 48 hours, extracellular parasites were washed away and T. cruzi – infected fibroblasts were harvested. Peptides presented on Major Histocompatibility Complex I (MHC-I) were isolated and analyzed by mass spectrometry using an ThermoScientific Orbitrap Eclipse instrument. Proteomes databases T. cruzi strain CL Brener (https://www.uniprot.org/proteomes/UP000002296) and C57BL/6 mus musculus (https://www.uniprot.org/proteomes/UP000000589) were used for peptide identification. In this dataset, experiment #1 and #2 represents raw data from mass spec analysis on peptides from T. cruzi – infected MC57G fibroblasts. Experiment #3 contains data from uninfected MC57G cells as control.