Chagas disease (CD) is a neglected tropical disease caused by Trypanosoma cruzi infection that, despite being discovered over a century ago, remains a public health problem, mainly in developing countries. Since T. cruzi can infect a wide range of mammalian host cells, parasite–host interactions may be critical to infection outcome. The intense immune stimulation that helps the control of the parasite’s replication and dissemination may also be linked with the pathogenesis and symptomatology worsening. Here, we discuss the findings that support the notion that excessive immune system stimulation driven by parasite persistence might elicit a progressive loss and collapse of immune functions. In this context, cellular stress and inflammatory responses elicited by T. cruzi induce fibroblast and other immune cell senescence phenotypes that may compromise the host’s capacity to control the magnitude of T. cruzi-induced inflammation, contributing to parasite persistence and CD progression. A better understanding of the steps involved in the induction of this chronic inflammatory status, which disables host defense capacity, providing an extra advantage to the parasite and predisposing infected hosts prematurely to immunosenescence, may provide insights to designing and developing novel therapeutic approaches to prevent and treat Chagas disease.

The intestinal lamina propria harbors a diverse repertoire of innate and adaptive immune cell populations, whose dynamic regulation is critical for maintaining mucosal immune homeostasis and for mounting effective inflammatory responses to infection or tissue injury. Accurate characterization of these populations in murine models requires reliable isolation of viable cells while preserving surface marker integrity. However, the heterogeneous cellular composition of the lamina propria poses significant technical challenges, and variability in isolation protocols has contributed to reproducibility issues across studies in mouse experimental models. In particular, enzymatic digestion can markedly affect cell viability and surface molecule expression, with outcomes depending on the choice of enzyme and the inclusion of additives such as deoxyribonuclease I (DNase I) and/or dithiothreitol (DTT). Standardization of isolation methods in the immunology field is therefore essential to ensure consistent recovery of viable immune cells suitable for downstream analyses. To promote reproducibility across laboratories, we introduce a minimal yet comprehensive reporting checklist, together with a systematic overview of the key experimental variables that govern the yield, viability, and phenotypic integrity of lamina propria leukocytes (LPLs) in mice.