Toxoplasma gondii (T. gondii) is the protozoan parasite that causes toxoplasmosis, a potentially fatal disease to immunocompromised patients, and which affects approximately 30% of the world’s population. Previously, we showed that purinergic signaling via the P2X7 receptor contributes to T. gondii elimination in macrophages, through reactive oxygen species (ROS) production and lysosome fusion with the parasitophorous vacuole. Moreover, we demonstrated that P2X7 receptor activation promotes the production of anti-parasitic pro-inflammatory cytokines during early T. gondii infection in vivo. However, the cascade of signaling events that leads to parasite elimination via P2X7 receptor activation remained to be elucidated. Here, we investigated the cellular pathways involved in T. gondii elimination triggered by P2X7 receptor signaling, during early infection in macrophages. We focused on the potential role of the inflammasome, a protein complex that can be co-activated by the P2X7 receptor, and which is involved in the host immune defense against T. gondii infection. Using peritoneal and bone marrow-derived macrophages from knockout mice deficient for inflammasome components (NLRP3−/−, Caspase-1/11−/−, Caspase-11−/−), we show that the control of T. gondii infection via P2X7 receptor activation by extracellular ATP (eATP) depends on the canonical inflammasome effector caspase-1, but not on caspase-11 (a non-canonical inflammasome effector). Parasite elimination via P2X7 receptor and inflammasome activation was also dependent on ROS generation and pannexin-1 channel. Treatment with eATP increased IL-1β secretion from infected macrophages, and this effect was dependent on the canonical NLRP3 inflammasome. Finally, treatment with recombinant IL-1β promoted parasite elimination via mitochondrial ROS generation (as assessed using Mito-TEMPO). Together, our results support a model where P2X7 receptor activation by eATP inhibits T. gondii growth in macrophages by triggering NADPH-oxidase-dependent ROS production, and also by activating a canonical NLRP3 inflammasome, which increases IL-1β production (via caspase-1 activity), leading to mitochondrial ROS generation.

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Objetivo: Descrever a estruturação e implementação de farmácia clínica na unidade de terapia intensiva. Métodos: Trata-se de estudo descritivo, cuja metodologia foi elaboração de fluxograma de atividades desenvolvidas pelo farmacêutico e formulários para facilitar acompanhamento farmacoterapêutico. Para confecção dos documentos foi realizada uma pesquisa bibliográfica e consulta a outros formulários de acompanhamento clínico da própria instituição e de outros serviços de farmácia clínica. Resultados: Foram criados documentos estruturantes: fluxograma do serviço de farmácia; formulário de acompanhamento farmacoterapêutico; check-list de avaliação da prescrição e check-list de acompanhamento farmacoterapêutico; modelo de notificação de intervenção farmacêutica; e, banco de dados para registro das intervenções a serem realizadas. Esses documentos contam com informações importantes para o acompanhamento diário do paciente. Conclusão: O método proposto auxilia no desenvolvimento de atividades clínicas, promovendo integração do farmacêutico à equipe multiprofissional, cuidado integral do paciente e segurança. Adaptação a formulário próprio garante a equipe conduzir acompanhamento de personalizado.