Estudo da modulação do metabolismo lipídico de células dendríticas humanas na infecção por Toxoplasma gondii / Study of modulating lipid metabolism in human dendritic cells infected with Toxoplasma gondii

A toxoplasmose é uma doença de alta prevalência no mundo, causada pelo Toxoplasma gondii, um parasita intracelular obrigatório. O T. gondii possui mecanismos de modulação do metabolismo do seu hospedeiro que garantem sua sobrevivência e a instalação da infecção crônica. Um dos tipos celulares mais permissivos à infecção e multiplicação do T. gondii são as células dendríticas (DC), paradoxalmente as células apresentadoras de antígeno mais eficazes, com capacidade de deflagrar uma resposta imune protetora eficaz e duradoura. Neste trabalho, estudamos a modulação do metabolismo lipídico de células dendríticas humanas na infecção por T. gondii. Mostramos que o parasita induziu a maturação da célula e um padrão misto de resposta inflamatória, com altas concentrações das citocinas pró-inflamatórias IL-6 e TNF-alfa e da citocina antiinflamatória IL-10. Após 3 horas de infecção, verificamos que T. gondii induziu a expressão gênica de ciclooxigenase-2 (COX-2). De forma importante, observamos por qRT-PCR que a infecção com T. gondii regulou positivamente a expressão do gene do receptor nuclear (RN) regulado por lipídios PPARgama, mas não do LXRbeta. Já a expressão do mRNA das moléculas envolvidas no transporte e estoque de lipídios, FABP4 e ADRP, alvos do PPARgama, e ABCA1, alvo do LXRbeta, foram aumentadas pela infecção de 3 horas com o parasita. Utilizando duas técnicas distintas (BODIPY e coloração com ósmio) avaliamos a biogênese de corpúsculos lipídicos (CL) após infecção com o T. gondii e constatamos que essas organelas não foram induzidas após 3 horas de infecção. Entretanto, após 24 horas, 90% das DC apresentaram CL e o número de CL por DC foi estatisticamente maior. Observamos a presença de CL em DC não infectadas pelo parasita, mostrando que a indução da biogênese de CL é um fenômeno parácrino, não dependente da infecção celular. Avaliamos também a importância do PPARgama na infecção por T. gondii, através do tratamento das DC com seu agonista ou antagonista. Após 3 horas, apenas os genes ADRP, FABP4 e ABCA1, alvos dos receptores nucleares, foram modulados. Por último, investigamos a influência do T. gondii na expressão das moléculas apresentadoras de antígenos lipídicos. Por citometria de fluxo, constatamos que não há alteração na expressão de membrana dessas moléculas. Contudo, por qRT-PCR, observamos que o T. gondii regula negativamente a expressão dos genes cd1d e cd1e. Em conclusão, mostramos que T. gondii foi capaz de regular positivamente o metabolismo lipídico das DC e negativamente as moléculas apresentadoras de lipídios CD1, sem a participação essencial de PPAR nesses processos.

Tratamento com bioterápico Candida albicans influencia a adesão de fungos a células Ma104 in vitro / Treatment with Candida albicans biotherapic influences in vitro fungal adhesion to Ma-104 cells

Oral candidiasis is an opportunist fungal infection in humans, mainly caused by Candida albicans. It occurs when the host presents an imbalance in the immune system and Candida spp., normally found in human flora, become able to develop the infection [1]. This disease is very common in HIV patients, and in all individuals that present immunossupression, such as patients treated with chemotherapy. Considering this scenario, the development of new medicines to treat oral candidiasis is mandatory.These results showed that the biotherapic did not present any citotoxicity, but was able to modify the morphological aspects of Ma-104 cells. Additionally, the interaction between host cells and ethilogic agent is directly influenced by biotherapic treatment, suggesting a promising antifungal potential of this medicine.(AU)

Cell alterations induced by a biotherapic for influenza / Cell alterations induced by a biotherapic for influenza

Introduction: Influenza viruses have been responsible for highly contagious acute respiratory illnesses with high mortality, mainly in the elderly, which encourages the development of new drugs for the treatment of human flu. The biotherapics are medicines prepared from biological products, which are not chemically defined. They are compounded following the homeopathic procedures indicated for infectious diseases with known etiology [1]. Aim: The purpose of the present study is to verify cellular alterations induced by a biotherapic prepared from the infectious influenza A virus. Methodology: This biotherapic was prepared for this study in the homeopathic potency of 30X according to the Brazilian Homeopathic Pharmacopeia [2]. The concentration of 10% was not cytotoxic to cells, as verified by neutral red assay. The cellular alterations observed in MDCK cells were analyzed by optical microscopy for the quantification of mitosis, nucleoli and lipid bodies. The mitochondrial activity was assessed by MTT assay and the phosphosfructokinase-1 (PFK-1) enzyme activity was analyzed on the MDCK cells treated for 5, 10 and 30 days. Macrophages J778.G8 were treated with this biotherapic to evaluate the immunostimulatory cytokine release. Results: The cellular alterations observed in MDCK cells were verified by optical microscopy. The number of lipid bodies present in MDCK cells stimulated for 10 days was significantly lower (p <0.05) when compared to controls. The biotherapic significantly increased (p <0.05) the number of mitosis and the mitochondrial activity of MDCK cells stimulated for 10 and 30 days. These changes were confirmed by a significant reduction (p <0.05) on the PFK-1 activity. These results suggest that the biotherapic was able to activate the Krebs cycle and pentosephosphate metabolism to the generation of amino acids and nucleotides, situations common to cells whose rate of mitosis is increased. The quantification of immunostimulatory cytokines by macrophages J774.G8 indicated that the tumor necrosis factor (TNF-?) production was higher (p <0.05) in the supernatant of the macrophages pre-treated with this biotherapic and infected with influenza virus, suggesting an activation of the macrophages by this biotherapic. Conclusion: This biotherapic is able to induce some cellular alterations, which show strong evidence that it might be a promising option for the human flu. New experiments are being developed to understand the mechanisms of action of this biotherapic.(AU)

Cell alterations induced by a biotherapic for influenza / Cell alterations induced by a biotherapic for influenza

Introduction: Influenza viruses have been responsible for highly contagious acute respiratory illnesses with high mortality, mainly in the elderly, which encourages the development of new drugs for the treatment of human flu. The biotherapics are medicines prepared from biological products, which are not chemically defined. They are compounded following the homeopathic procedures indicated for infectious diseases with known etiology [1]. Aim: The purpose of the present study is to verify cellular alterations induced by a biotherapic prepared from the infectious influenza A virus. Methodology: This biotherapic was prepared for this study in the homeopathic potency of 30X according to the Brazilian Homeopathic Pharmacopeia [2]. The concentration of 10% was not cytotoxic to cells, as verified by neutral red assay. The cellular alterations observed in MDCK cells were analyzed by optical microscopy for the quantification of mitosis, nucleoli and lipid bodies. The mitochondrial activity was assessed by MTT assay and the phosphosfructokinase-1 (PFK-1) enzyme activity was analyzed on the MDCK cells treated for 5, 10 and 30 days. Macrophages J778.G8 were treated with this biotherapic to evaluate the immunostimulatory cytokine release. Results: The cellular alterations observed in MDCK cells were verified by optical microscopy. The number of lipid bodies present in MDCK cells stimulated for 10 days was significantly lower (p <0.05) when compared to controls. The biotherapic significantly increased (p <0.05) the number of mitosis and the mitochondrial activity of MDCK cells stimulated for 10 and 30 days. These changes were confirmed by a significant reduction (p <0.05) on the PFK-1 activity. These results suggest that the biotherapic was able to activate the Krebs cycle and pentosephosphate metabolism to the generation of amino acids and nucleotides, situations common to cells whose rate of mitosis is increased. The quantification of immunostimulatory cytokines by macrophages J774.G8 indicated that the tumor necrosis factor (TNF-?) production was higher (p <0.05) in the supernatant of the macrophages pre-treated with this biotherapic and infected with influenza virus, suggesting an activation of the macrophages by this biotherapic. Conclusion: This biotherapic is able to induce some cellular alterations, which show strong evidence that it might be a promising option for the human flu. New experiments are being developed to understand the mechanisms of action of this biotherapic.