RESUMEN
American trypanosomiasis has long been a neglected disease endemic in LatinAmerica, but congenital transmission has now spread Chagas disease to cause a global health problem. As the early stages of the infection of placental tissue and the vertical transmission by Trypanosoma cruzi are still not well understood, it is important to investigate the relevance of the first structure of the placental barrier in chorionic villi infection by T. cruzi during the initial stage of the infection. Explants of human chorionic villi from healthy pregnant women at term were denuded of their syncytiotrophoblast and co-cultured for 3h, 24h and 96h with 800,000 trypomastigotes (simulating acute infection). T. cruzi infected cells were identified by immunohistochemistry for cytokeratin-7 (+cytotrophoblast) and CD68 (+macrophages), and the infection was quantified. In placental tissue, the parasite load was analyzed by qPCR and microscopy, and the motile trypomastigotes were quantified in culture supernatant. In denuded chorionic villous, the total area occupied by the parasite (451.23µm2, 1.33%) and parasite load (RQ: 87) was significantly higher (p<0.05) than in the entire villous (control) (5.98µm2, 0.016%) (RQ:1) and with smaller concentration of nitric oxide. Stromal non-macrophage cells were infected as well as cytotrophoblasts and some macrophages, but with significant differences being observed. The parasite quantity in the culture supernatant was significantly higher (p<0.05) in denuded culture explants from 96h of culture. Although the human complete chorionic villi limited the infection, the detachment of the first structure of the placenta barrier (syncytiotrophoblast) increased both the infection of the villous stroma and the living trypomastigotes in the culture supernatant. Therefore structural and functional alterations to chorionic villi placental barrier reduce placental defenses and may contribute to the vertical transmission of Chagas.
Asunto(s)
Enfermedad de Chagas/transmisión , Vellosidades Coriónicas/parasitología , Transmisión Vertical de Enfermedad Infecciosa , Trypanosoma cruzi/metabolismo , Antígenos CD/inmunología , Antígenos de Diferenciación Mielomonocítica/inmunología , Técnicas de Cocultivo , Femenino , Humanos , Queratina-7/inmunología , Óxido Nítrico , Placenta/parasitología , Reacción en Cadena de la Polimerasa , EmbarazoRESUMEN
The aim of the work was to analyze the susceptibility of the placental syncytiotrophoblast (STB) and cytotrophoblast (CTB) cells to infection by the causal agent of congenital Chagas' disease, Trypanosoma cruzi, and the possible parasite route for placental invasion. Monolayers of CTB and STB and VERO as control cells were used. The infection of STB was significantly lower that of the CTB and Vero cells (p < 0.05) which coincided with a significantly increased mortality of parasite cells in the culture medium and trypanocidal levels of nitric oxide. We conclude that the syncytiotrophoblast, the first placental barrier, is the main barrier of the chorionic villous that limits the infection by T. cruzi. This work opens the possibility of a new mechanism for placental infection when there are discontinuities in the first placental barrier.
Asunto(s)
Enfermedad de Chagas/parasitología , Trofoblastos/parasitología , Trypanosoma cruzi/fisiología , Animales , Células Cultivadas , Enfermedad de Chagas/congénito , Enfermedad de Chagas/patología , Enfermedad de Chagas/transmisión , Chlorocebus aethiops , Medios de Cultivo Condicionados/metabolismo , Susceptibilidad a Enfermedades , Femenino , Humanos , Transmisión Vertical de Enfermedad Infecciosa , Óxido Nítrico/metabolismo , Embarazo , Complicaciones Parasitarias del Embarazo/parasitología , Trofoblastos/citología , Trofoblastos/patología , Trypanosoma cruzi/crecimiento & desarrollo , Trypanosoma cruzi/aislamiento & purificación , Células VeroRESUMEN
UNLABELLED: We hypothesize that a sustained infection of Trypanosoma cruzi into placental tissue might be diminished. Human placental chorionic villi and VERO cells as controls were co-cultured with T. cruzi. Parasites occupied 0.0137% at 3h, 0.0224% (24h), and 0.0572% (72 h) of the total chorionic villi area analyzed and some few placental samples were negative to parasite DNA, whereas 52% of VERO cells were infected at 3h and parasites occupied 0.57%, at 24h the parasite area was of 2.78% and at 72 h was of 3.32%. There were no live parasites in placenta-T. cruzi culture media at 72 h of co-culture. There were significantly increased dead parasites when T. cruzi was treated with unheated culture media coming from placental explants and fewer dead parasites when pre-heated culture media were employed. CONCLUSION: Low productive infection by T. cruzi into placental tissue associated with no viable parasites in the culture media partially due to placental thermo labile substances.