Vacinação de camundongos com células dendríticas derivadas de células tronco contendo o gene Rho1-GTPase de Schistosoma mansoni.

Abstract

Schistosomiasis remains one of the worlds most prevalent diseases. Several studies have been made aiming the development of a vaccine capable of protecting against Schistosoma mansoni infection or limiting the development of females as well as suppressing of the egg production. However, many years of searching failed to identify a vaccine candidate. In different models of disease, it has been demonstrated that dendritic cells (DCs) can serve as vaccine carriers, mediating protection against various types of pathogens. The studies of ex vivo manipulation requirements of DCs may lead to the design of vaccines that induce protective immunity to infections by appropriate targeting of DCs in vivo. Objective: To evaluate the immune reply of mice vaccinated with modified stem cell derived in dendritic cells containg the gene Rho1-GTPase of Schistosoma mansoni. Methods: Dendritics cells (DC) were obtained from the maturation of bone marrow stem cells with conditional mediumcontaining GM-CSF and further characterized by flow cytometry and confocal microscope. After amplification by PCR, the fragment of Rho1-GTPase was inserted in a vector of expression in mammals cells, GFP fusion TOPO TA, and then transfected in dendritic cells. These cells were used in the vaccination of mice submitted to an infection challenge with cercariae. Results and conclusion: Analysis of the bone marrow derived cell population showed that these cells expressed the distinctive DC surface marker CD11c, as well as the major histocompatibility complex (MHC) classes I and II molecules. Dendritic cells were visualized on a Zeiss confocal microscope to evaluate phagocytic activity, shown by the uptake of fluorescent-labeled latex beads and the typical morphology. Once cloned, analyzed, and transfected, the GFP fusion protein was expressed by DC. The expression of Rho1- GTPase was observed by an efficient transfection on DC and the best approach was observed from GeneJuice compared with nanoparticles and Effectene. Analysis of protection against S. mansoni revealed that altered DC induced 56% of protection, relatively higher than that conferred only by the same encoded protein, 51%. We conclude that the dendritic cell is avaluable tool to control schistosomiasis and the present results represent a breakthrough in the production of new vaccine approaches.