Isolation and Phenotypic Characterization of Inflammatory Cells from Clinical Samples: Purification of Macrophages from Trypanosoma cruzi-Infected Hearts.

Abstract

Trypanosoma cruzi, the causal agent of chronic Chagas cardiomyopathy, exhibits an important tropism for cardiac tissue. In consequence, T. cruzi experimental infection represents a unique model to study cardiac macrophage behavior and effector functions during either acute or chronic immune response. In this chapter we describe a protocol to isolate immune cells from T. cruzi-infected murine cardiac tissue and to determine the percentage, absolute number, phenotype, and functionality of monocytes and macrophages by using flow cytometry. Moreover, we describe the parameters to discriminate between resident and infiltrating mononuclear phagocytic cells within infected hearts. The investigations in this field will provide mechanistic insights about the roles of these innate immune cells in the context of a clinically relevant target tissue.