Avaliação do perfil de ativação de células dendríticas através da análise de moléculas co-estimulatórias e citocinas em pacientes com lesões intraepiteliais cervicais e câncer cervical invasivo

Abstract

The present study aimed to investigate the activation profile of monocyte-derived dendritic cells from patients with cervical intraepithelial lesions of varying degrees, undergoing three different activation protocols, in order to verify that the staging of cervical injury can influence on the process of differentiation of these cells and if different activation protocols could be result in dendritic cells with increased maturation phenotype. A total of 83 women were included in this study, of these were collected peripheral blood to obtained mononuclear cells witch were subjected to three different differentiation protocols. In protocol I was used GM-CSF, IL-4 and TNF-α, for the differentiation of monocytes into dendritic cells. In protocol II, monocytes were differentiated by the stimulation with GM-CSF, IL-4 and TNF-α and then co-stimulated with non-adherent cells stimulated with LPS. In protocol III, mononuclear cells from peripheral blood were stimulated with GM-CSF, IL-4 and TNF-α for the differentiation of dendritic cells and then co-stimulated with mononuclear cells stimulated by LPS. The differentiated dendritic cells were evaluated for the expression of surface markers and co-stimulatory molecules and the culture supernatant was assessed for the presence of cytokines. Our study revealed that the process of differentiation and activation of dendritic cells differentiated from peripheral blood monocytes of patients with invasive cervical cancer is impaired, since we observed a significant decrease in expression of CD209. The results observed from the comparison between the protocols used also revealed that stimulation with LPS is capable of inducing a better differentiation of dendritic cells activated with TNF-α, obtained from patients with LSIL and HSIL. In addition, we also observed that LPS is capable of inducing a significant increase of CD86 and HLA-DR expression and of IL-12 production, are so important for the differentiation of Th1 anti-tumor immune response profile. The presence of non-adherent cells in the culture of dendritic cells appears to be essential for their differentiation, since their removal on protocol II, induced a significant reduction of the expression of surface markers CD11c, CD86 and HLA-DR, and IL-12, TNF-α and IL-10 synthesis. Thus, more studies are required to understand the effects of non-adherent cells and stimulation with TNF-α associated with LPS on the process of differentiation of dendritic cells.