ABSTRACT
Congenital toxoplasmosis is a parasitic disease that occurs due vertical transmission of the protozoan Toxoplasma gondii (T. gondii) during pregnancy. The parasite crosses the placental barrier and reaches the developing brain, infecting progenitor, glial, neuronal and vascular cell types. Although the role of Radial glia (RG) neural stem cells in the development of the brain vasculature has been recently investigated, the impact of T. gondii infection in these events is not yet understood. Herein, we studied the role of T. gondii infection on RG cell function and its interaction with endothelial cells. By infecting isolated RG cultures with T. gondii tachyzoites, we observed a cytotoxic effect with reduced numbers of RG populations together with decrease neuronal and oligodendrocyte progenitor populations. Conditioned medium (CM) from RG control cultures increased ZO-1 protein levels and organization on endothelial bEnd.3 cells membranes, which was impaired by CM from infected RG, accompanied by decreased trans-endothelial electrical resistance (TEER). ELISA assays revealed reduced levels of anti-inflammatory cytokine TGF-ß1 in CM from T. gondii-infected RG cells. Treatment with recombinant TGF-ß1 concomitantly with CM from infected RG cultures led to restoration of ZO-1 staining in bEnd.3 cells. Congenital infection in Swiss Webster mice led to abnormalities in the cortical microvasculature in comparison to uninfected embryos. Our results suggest that infection of RG cells by T. gondii negatively modulates cytokine secretion, which might contribute to endothelial loss of barrier properties, thus leading to impairment of neurovascular interaction establishment.
Subject(s)
Cell Differentiation , Cerebral Cortex/blood supply , Endothelial Cells/parasitology , Ependymoglial Cells/parasitology , Microvessels/parasitology , Neurovascular Coupling , Toxoplasma/pathogenicity , Toxoplasmosis, Cerebral/parasitology , Toxoplasmosis, Congenital/parasitology , Animals , Cell Line , Disease Models, Animal , Electric Impedance , Endothelial Cells/metabolism , Endothelial Cells/pathology , Ependymoglial Cells/metabolism , Ependymoglial Cells/pathology , Mice, Inbred C57BL , Microvessels/metabolism , Microvessels/pathology , Tight Junctions/metabolism , Tight Junctions/parasitology , Tight Junctions/pathology , Toxoplasmosis, Cerebral/metabolism , Toxoplasmosis, Cerebral/pathology , Toxoplasmosis, Congenital/metabolism , Toxoplasmosis, Congenital/pathology , Transforming Growth Factor beta1/metabolism , Zonula Occludens-1 Protein/metabolismABSTRACT
Toxoplasma protein disulfide isomerase (PDI) is a 52 KDa thioredoxin of interest because have a great immunogenicity for humans. We cloned and produced a recombinant protein (recTgPDI) used to test its effect during infection to different human cell lines (epithelial and retinal). We also determine if there were differences in gen expression during in vitro infection. Expression of the gen was lower after entry into the host cells. PDI's inhibitors bacitracin and nitroblue tetrazolium reduced the percent of infected cells and small amounts of recTgPDI proteins interfered with the invasion step. All these results support a role of Toxoplasma PDI during the first steps of infection (adhesion and invasion). Toxoplasma PDI is a protein linked to early steps of invasion, it would be of importance to identify the host proteins substrates during invasion steps.
