Generation of T. cruzi-specific primary CD4⁺ T cell lines from peripheral blood mononuclear cells isolated from chagas disease patients

Human CD8⁺ and CD4⁺ T cell lines and clones are valuable tools to explore the role of these cells in the context of diseases, especially in cases in which the main underlying actor is the immune response, like Chagas disease. These cell lines and clones provide a good experimental system to address the phenotypic and functional features of specific T cell subpopulations and furthermore settle the framework necessary for analyzing their antigen/peptide specificity. This chapter details a culture method for the establishment of T. cruzi-specific memory T cell lines from mononuclear cells isolated from Chagas disease patients’ peripheral blood. The presented protocol comprises (1) enrichment of memory CD4⁺ T cells, (2) stimulation with parasite lysate, (3) evaluation of specificity, and (4) expansion and maintenance of specific T cell lines.