O uso da PCR em tempo real para o estudo da carga parasitária e dos níveis transcricionais durante a infecção experimental por Trypanosoma cruzi

Trypanosoma cruzi is causative agent of Chagas disease, one of most neglected tropical diseases. Estimated that about 11 million people worldwide are infected by T. cruzi and about 6 to 7 million people are at risk in endemic areas. During the process of invasion of host and parasite interact enabling signal transduction and gene expression modulation in response to invasion. The diversity of activated proteins and pathways to repair the damage by disruption of the plasma membrane interest to us and thus present study developed a new form of detection and quantitation by polymerase chain reaction in real time (qPCR) of parasitic load T. cruzi and quantified transcriptional levels relative (RT-qPCR) of dysferlin, Sphingomyelin acid esferase (ASM), transcription factor EB (TFEB) Galectins 1 and 3 and Annexin A2. This study demonstrated that quantification by real time PCR using primers P21fw and P21rv was specific and sensitive for detection of T. cruzi in vivo and in vitro, as well as transcriptional levels of genes related to cytoskeletal organization and repair plasma membrane are modulated in response to damage generated by parasite.

Análise do perfil imunológico e citohistológico durante a transmissão vertical de Trypanosoma cruzi em diferentes estágios da infecção experimental

Trypanosma cruzi is the causative agent of Chagas disease. This trypanosomiasis has become a global public health problem due to migration of Latin Americans to non-endemic countries. In Latin America with the succesful implementation of control domiciliated vector infestation and blood transfusion, the importance of congenital transmission has recently increased. Considering the tight regulation of immune system during gestation, we aimed to investigate the changes in the immune system caused by T.cruzi infection in the gestation outcome. T cruzi G and Y strain were used to infect female BALB/c mice before or after mating with non-infected male mice. The presence of vaginal plug was used as indicative of mating. Females were euthanized 8 days after confirmation of vaginal plug. We used three female control groups, only infected, only infected and non-infected and non-pregnant females. Two groups were infected before mating and other two were infected 4 days after confirmation of vaginal plug. The uterus and spleen were collected to immunochemistry, qPCR, immunofluorescence and cytokine analysis. Our results showed that despite the MMP’s identification being similarly among groups, T.cruzi higher virulent strain can impaire gestation outcome prior mating; the infection also increased cytokines like IFN-γ, IL-1β and IL-4; and leucocytes in uterine environment was altered, responding locally to systemic changes caused by T.cruzi infection. In conclusion this work suggests that T.cruzi infection can impaire gestation outcome and local response to sistemic infection was able to control the infection allowing pregnancy development in some conditions.