Gene expression network analyses during infection with virulent and avirulent Trypanosoma cruzi strains unveil a role for fibroblasts in neutrophil recruitment and activation

Abstract

Chagas disease is caused by Trypanosoma cruzi , a protozoan parasite that has a heterogeneous population composed of a pool of strains with distinct characteristics, including variable levels of virulence. In previous work, transcriptome analyses of parasite genes after infection of human foreskin fibroblasts (HFF) with virulent (CL Brener) and non-virulent (CL-14) clones derived from the CL strain, revealed a reduced expression of genes encoding parasite surface proteins in CL-14 compared to CL Brener during the final steps of the intracellular differentiation from amastigotes to trypomastigotes. Here we analyzed changes in the expression of host genes during in vitro infection of HFF cells with the CL Brener and CL-14 strains by analyzing total RNA extracted from cells at 60 and 96 hours post-infection (hpi) with each strain, as well as from uninfected cells. Similar transcriptome profiles were observed at 60 hpi with both strains compared to uninfected samples. However, at 96 hpi, significant differences in the number and expression levels of several genes, particularly those involved with immune response and cytoskeleton organization, were observed. Further analyses confirmed the difference in the chemokine/cytokine signaling involved with the recruitment and activation of immune cells such as neutrophils upon T . cruzi infection. These findings suggest that infection with the virulent CL Brener strain induces a more robust inflammatory response when compared with the non-virulent CL-14 strain. Importantly, the RNA-Seq data also exposed an unexplored role of fibroblasts as sentinel cells that may act by recruiting neutrophils to the initial site of infection. This role for fibroblasts in the regulation of the inflammatory response during infection by T . cruzi was corroborated by measurements of levels of different chemokines/cytokines during in vitro infection and in plasma from Chagas disease patients as well as by neutrophil activation and migration assays.

The Complete Mitochondrial DNA of Trypanosoma cruzi: Maxicircles and Minicircles

“…Besides, some researchers have suggested that both the parasite and host genetic variability could be the causes of differential clinical manifestations of Chagas disease ( Manoel-Caetano Fda and Silva, 2007 ). Also, transcriptomic analysis revealed differences between virulent and non-virulent strains ( Belew et al., 2017 ; Oliveira et al., 2020 ). Indeed, some T. cruzi strains highly differ in pathogenicity.…”

LncRNA: A Potential Target for Host-Directed Therapy of Candida Infection

“…Our preliminary studies reported that fibroblasts could secrete CX3CL1 to effectively prevent the adhesion and invasion of Candida [ 73 ]. Fibroblasts may also play indirect antifungal roles by recruiting and altering the functional status of immune cells through transforming growth factor-β1 (TGF-β1), granulocyte colony-stimulating factor (G-CSF), or CCL2 [ 74 , 75 ]. In addition, the crosstalk between fibroblasts and immune cells was reported in cancer [ 76 ].…”

Serum from Laron Mice Partially Protects against <i>Trypanosoma Cruzi</i> Infection

“…Previously RNA-Seq data [39][40][41] from human foreskin fibroblast cells infected with T. cruzi strain CL Brenner and Sylvio at 96 hours and 72 hours post-infection (hpi) were selected to generate a heat map. Four differentially expressed genes (GHR, IGF-1, IGF-1R, IGFBP3) RNA-seq data were extracted and plotted.…”

“…The heat map generated, using previous RNA-Seq data [39][40][41] from differentially expressed genes, showed that during T. cruzi infection in human foreskin fibroblast cells GHR gene expression is downregulated (Figure 1a). Importantly, these data show that different T. cruzi strains (Sylvio, CL Brenner), at different times of infections (96 and 72 hpi), consistently downregulate GHR expression.…”