Effect of adoptive transfer of spleen cells or CD4+ T cells stimulated with ArtinM during infection with Paracoccidioides brasiliensis

Abstract

Paracoccidioidomycosis (PCM) is caused by the fungus Paracoccidioides brasiliensis, that is dimorphism in a temperature dependent manner. The disease in humans is characterized by the transformation of the mycelial form of the fungus (infectious) to the yeast form (pathogenic). The establishment of Th1 response mediated by IL-12 is required for the development of protective immunity against PCM. In the experimental PCM, the prophylactic or therapeutic administration of ArtinM confers resistance to PCM throught of the immune response to the Th1 profile. The immunomodulatory activity induced by ArtinM promotes the protection against other intracellular pathogens, such as: Leishmania amazonensis, Leishmania major, Neospora caninum and Candida albicans. ArtinM is extracted from seeds of Artocarpus heterophyllus and exerts some biological activities: activates neutrophils, promotes the degranulation of mast cells, induces the production of IL-12 on antigen-presenting cells (APCs), activates spleen cells and CD4+ T cells. These activities result in a pattern of adequate immune response favorable to against intracellular pathogens. To investigate the contribution of lymphocytes activated by ArtinM against PCM, we propose to analyze the ability of spleen cells or CD4+ T cells after incubation with ArtinM. For this, we will realize the adoptive transfer these cells and evaluate the effect in the course of the disease. Thus, BALB/c mice will be infected with P. brasiliensis (1x106yeast/mice). After 10 days of infection, these mice will receive spleen cells or CD4+ T cells obtained from BALB/c that were stimulated ex vivo with ArtinM (625ng/mL) during 24 hours before adoptive transfer. After 30 days of infection the mice will be sacrificed and their blood and organs (spleen, liver and lung) will be collected. We will be analyzed the pro-inflammatory and anti-inflammatory cytokines in the serum and extract of the organs. Moreover, the organs will be verified by histopathology, analysis of fungal burden and quantified the frequency of granulomas. The results will be compared between the infected and non-infected mice that received by adoptive transfer the spleen cells or CD4+ T cells stimulated or not with ArtinM.