Expression data from human primary fibroblasts, endothelial and smooth muscle cells infected with Trypanosoma cruzi

Trypanosoma cruzi is an obligate intracellular protozoan parasite that causes human Chagas’ disease, a leading cause of heart failure in Latin America. Using Affymetrix oligonucleotide arrays we screened phenotypically diverse human cells (foreskin fibroblasts, microvascular endothelial cells and vascular smooth muscle cells) for a common transcriptional response signature to T. cruzi. A common feature was a prominent type I interferon response, indicative of a secondary response to secreted cytokines. Using transwell plates to distinguish cytokine-dependent and -independent gene expression profiles in T. cruzi-infected cells, a core cytokine-independent response was identified in fibroblasts and endothelial cells that featured metabolic and signaling pathways involved in cell proliferation, amino acid catabolism and response to wounding. Significant downregulation of genes involved in mitotic cell cycle and cell division predicted that T. cruzi infection impedes cell cycle progression in the host cell. Primary human foreskin fibroblasts and human microvascular endothelial cells were grown in transwell dishes to reach 80% confluency. The cells in the bottom of the transwell were infected with T. cruzi for 24 hours, while the cells on the top of the transwel were not infected but exposed to the same culture medium bathing the infected cells. Transwells containing uninfected cells in both levels were also included as controls. Vascular smooth muscle cells were omitted from the transwell experiments due to difficulties obtaining adequate RNA from cells plated on the top chamber. Instead they were cultred in Petri dishes and mock/infected with T. cruzi for 24 or 48 hours.Total RNA was extracted and gene expression levels analyzed with Affymetrix microarrays. Two independent biological replicates were included for each type of treatment.