Canine Macrophage DH82 Cell Line As a Model to Study Susceptibility to Trypanosoma cruzi Infection

Abstract

Trypanosoma cruzi is an obligatory intracellular protozoan parasite, and it is the etiolo gical agent of Chagas' disease that is endemic in the Americas. In addition to humans, a wide spectrum of mammals can be infected by T. cruzi, including dogs. Dogs develop acute and chronic disease, similar to human infection. T. cruzi can infect almost all cell types and after cell invasion, the metacyclics trypomastigotes localize in the cytoplasm, where they transform into amastigotes, the replicative form of T. cruzi in mammals. After amastigote multiplication and differentiation, parasites lyse host cells and spread through the body by blood circulation. In this work, we evaluated the in vitro ability of T. cruzi to infect a canine macrophage cell line DH82 compared with RAW264.7, a murine tissue culture macrophage. Our results have shown that the T. cruzi is able to infect, replicate and differentiate in DH82 cell line. We observed that following treatment with LPS and IFNγ DH82 cells were more resistant to infection and that resistance was not related reactive oxygen species production in our system. In this study, we also found that DH82 cells became more susceptible to T. cruzi infection when cocultured with apoptotic cells. The analysis of cytokine production has showed elevated levels of the TGFβ, IL10, and TNFα produced by T. cruziinfected canine macrophages. Additionally, we demon strated a reduced expression of the MHC class II and CD80 by infected DH82 cell line.

P2Y2 and P2X4 Receptors Mediate Ca2+ Mobilization in DH82 Canine Macrophage Cells

“…To date, the DH82 canine macrophage cell line has primarily been utilized as a model of viral and protozoan infection [23][24][25], such as for the study of canine distemper and its oncolytic potential [55][56][57]. Although studies have described the expression of functional P2 receptors in human or rodent macrophages and macrophage cell lines [8,10,11,34], studies directly investigating P2 receptors in canine macrophages have been absent.…”

Evaluation of an autologous cancer vaccine for the treatment of metastatic canine hemangiosarcoma: a preliminary study

“…Specifically, the extent of major histocompatibility complex II (MHC-II) and CD80 co-stimulatory molecules were up-regulated in response to incubation with vaccine or relevant controls. These signals are traditional markers of antigen-presentation and are correlated to the ability to stimulate T-cell activation in vivo [ 43 – 45 ]. Notably, the MHC-II signal is indicative of helper T-cell activation, not cytotoxic T-cell activation [ 46 ]; at this time, no validated canine-specific antibody is commercially available for MHC-I quantification, a marker of cytotoxic T-cell activation, via flow cytometry.…”

Efferocytosis and Outside-In Signaling by Cardiac Phagocytes. Links to Repair, Cellular Programming, and Intercellular Crosstalk in Heart

“…Another important feature of experimental infection with T. cruzi is the massive increase in apoptotic, activation-induced cell death in CD4 + T lymphocytes ( 21 ). Phagocytosis of these apoptotic lymphocytes by macrophages results in macrophage secretion of TGF-β leading to suppressive TGF-β signaling and increased growth of T. cruzi in the macrophage ( 22 , 23 ). Interestingly, in patients with cardiac clinical forms of Chagas disease, there is an increase in the expression of CCR5 on CD4 + T cells, which controls leukocyte migration into the inflamed heart ( 24 ), and while CCR5 expression is required during the acute phase for protection against experimental T. cruzi infection in mice, it is dispensable for the chronic phase of infection ( 25 ).…”