Correlation of cGAS, STING, INF-α and INF-β gene expression with Zika virus kinetics in primary culture of microglia and neurons from BALB/c mice

Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Biologia Molecular. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Biologia Molecular. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Biologia Molecular. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Microscopia Eletrônica. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Biologia Molecular. Ananindeua, PA, BrasilPattern recognition receptors participate in the innate immune response. Among PRRs, the cGAS/STING pathway is known to detect cytosolic DNA and cyclic dinucleotides, but it’s also important in RNA virus infection. We aimed to evaluate the gene expression of some important genes of cGAS/STING pathway and to correlate this expression with Zika virus kinetics in mice microglia and neurons. Cells were infected by MOI = 1.0. Indirect immunofluorescence, plaque titration of supernatant, extraction, and quantification of total intracellular RNA, RT-qPCR and Western blotting were performed. Plaque titration profile in microglia and neurons was similar, including higher titers of plaque forming units at 24, 48, 72 and 96 hpi, respectively. ZIKV kinetics evaluated by RT-qPCR was similar in both cells, with highest viral titers at 48, 72, 24 and 96 hpi, respectively. Expression profile of cGAS, STING, INF-α and INF-β was quite different between the cells, including gene suppression, as observed for cGAS in neurons. Our results showed a differentiated expression profile of cGAS/STING pathway genes in mice microglia and neurons, which can be explained by the different mechanisms that ZIKV uses to bypass the immune response of these cells. Furthermore, each cell type responds differently to combat the viral infection

Ultrastructural aspects of hemocytes from Biomphalaria glabrata Say (1818) (Gastropoda: Planorbidae) analysed with transmission eletronic microscopy

Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Os hemócitos do caramujo Biomphalaria glabrata, um importante transmissor do trematódeo Schistosoma mansoni no Brasil, foram coletados de especimens na região Bragantina, localizada a oeste do estado do Pará. Os hemócitos foram examinados por meio de microscopia eletrônica de transmissão. As células foram fixadas pelo método de rotina com o uso do tampão PHEM (PIPES, HEPES, EGTA e Magnésio). Foram descritos os aspectos ultra-estruturais celulares como inclusões citoplasmáticas limitadas por membranas, mitocôndrias, retículos endoplasmáticos e outros. As observações mostram que esse tampão possui a propriedade de preservação do citoesqueleto celular, apresentando bons resultados na preservação das estruturas dos hemócitos e suas organelas.The blood cells of the pulmonate snail Biomphalaria glabrata from the region of Bragantina in the state of Pará, an important vector of the trematode Schistosoma mansoni in Brazil, were collected and fixed by routine method with PHEM buffer and examined with transmission electron microscopy. Ultrastructural cellular aspects like cytoplasmic inclusions, mitochondrion, vesicles and others organelles are described. The buffer used resulted in good preservations of the hemocytes and their organelles

The fine structure of the axostyle and its associations with organelles in Trichomonads

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ), Programa de Núcleos de Excelência (PRONEX) and Associação Universitária Santa Úrsula (AUSU).Universidade Santa Ursula. Rio de Janeiro, RJ, Brazil / Universidade Federal do Rio de Janeiro. Instituto de Biofísica Carlos Chagas Filho. Rio de Janeiro, RJ, Brazil.Ministério da Saúde. Fundação Nacional de Saúde. Instituto Evandro Chagas. Belém, PA, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Biofísica Carlos Chagas Filho. Rio de Janeiro, RJ, Brazil / Universidade Federal. Rio de Janeiro, RJ, Brazil.The fine structure of the axostyle in the protists Tritrichomonas foetus and Monocercomonas sp is described using transmission electron microscopy after quick-freezing techniques and immunocytochemistry. The axostyle microtubules presents a lateral projection formed by two protofilaments in addition to the 13 protofilaments normally found in microtubules. The axostyle is associated with other cell structures such as hydrogenosomes, endoplasmic reticulum, sigmoid filaments and glycogen particles. The microtubules of the pelta-axostylar system are connected to each other by bridges regularly spaced with an interval of 9 nm. Labeling of the axostyle was observed after cell incubation with monoclonal antibodies recognizing α-tubulin and acetylated-tubulin

Trophozoites of Giardia lamblia may have a Golgi-like structure

Universidade Estadual do Norte Fluminense. Centro de Biociências e Biotecnologia. Laboratório de Biologia Celular e Tecidual. Campos dos Goytacazes, RJ, Brazil / Universidade Federal do Rio de Janeiro. Instituto de Biofísica Carlos Chagas. Programa de Biologia Celular e Parasitologia. Laboratório de Ultraestrutura Celular Hertha Meyer. Rio de Janeiro, RJ, Brazil.Universidade Estadual do Rio de Janeiro. Departamento de Histologia e Embriologia. Rio de Janeiro, RJ, Brazil.Ministério da Saúde. Fundação Nacional de Saúde. Instituto Evandro Chagas. Laboratório de Microscopia Eletrônica. Belém, PA, Brasil / Universidade Federal do Rio de Janeiro. Instituto de Biofísica Carlos Chagas. Programa de Biologia Celular e Parasitologia. Laboratório de Ultraestrutura Celular Hertha Meyer. Rio de Janeiro, RJ, Brazil.Universidade Estadual do Norte Fluminense. Centro de Biociências e Biotecnologia. Laboratório de Biologia Celular e Tecidual. Campos dos Goytacazes, RJ, Brazil / Filho, Universidade Federal do Rio de Janeiro. Instituto de Biofísica Carlos Chagas. Programa de Biologia Celular e Parasitologia. Laboratório de Ultraestrutura Celular Hertha Meyer. Rio de Janeiro, RJ, Brazil.Trophozoites of the primitive protozoan Giardia lamblia have been considered as cells which do not present the Golgi complex. Using C6-NBD ceramide, which has been shown to label the Golgi complex of mammalian cells, labelling of the perinuclear region of G. lamblia was observed by confocal laser scanning microscopy. Transmission electron microscopy of thin sections and of replicas of freeze-fractured cells revealed the presence of concentric perinuclear membranes resembling the Golgi complex. ß 1999 Federation of European Microbiological Societies

Granulocyte macrophage colony-stimulating factor alone reduces Toxoplasma gondii replication in microglial culture by superoxide and nitric oxide, without IFN-γ production: a preliminary report

Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil / Federal University of Pará. Postgraduate Program in Neuroscience and Cell Biology. Belém, PA, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Microscopia eletrônica. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Microscopia eletrônica. Ananindeua, PA, Brasil.Federal University of Pará. Institute of Biological Sciences. University Hospital João de Barros Barreto. Laboratory of Investigations in Neurodegeneration and Infection. Belém, PA, Brazil.State University of North Fluminense. Center of Biosciences and Biotechnology. Laboratory of Cell and Tissue Biology. Goytacazes, RJ, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Microscopia eletrônica. Ananindeua, PA, Brasil.In vitro studies have demonstrated that GM-CSF in combination with other stimulatory factors induces a microbicidal response that control T. gondii infection. We assessed whether GM-CSF alone can control T. gondii replication in murine microglial cultures. Microglia were collected and cultured with or without GM-CSF and the half of each group was infected with T. gondii. We determined the T. gondii infectivity, cytokines levels, NO and superoxide detection. GM-CSF alone primes microglia, which after infection induces the production of TNF-α and IL-6, leading to NO and superoxide production, without any stimulus from IL-12p70 and IFN-γ

Ultrastructural study of the gametocytes and merogonic stages of fallisia audaciosa (Haemosporina: Garniidae) that infect neutrophils of the lizard Plica umbra (Reptilia: Iguanidae)

Universidade Federal do Pará. Centro de Ciências Biológicas. Departamento de Patologia. Belém, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Belém, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Belém, PA, Brasil.Universidade Estadual do Norte Fluminense. Centro de Biociências e Biotecnologia. Laboratório de Biologia Celular e Tecidual. Campos de Goytacazes, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Biofísica Carlos Chagas Filho. Laboratório de Ultraestrutura Celular Hertha Meyer. Rio de Janeiro, RJ, Brasil.Little is known regarding the ultrastructure of the genus Fallisia (Apicomplexa: Haemosporina: Garniidae). This report describes the fine structure of some developmental stages of Fallisia audaciosa that infect neutrophils in the peripheral blood of the Amazonian lizard Plica umbra (Reptilia: Iguanidae). The parasites lie within a parasitophorous vacuole and exhibit the basic structures of members of the Apicomplexa, such as the pellicle and the cytostome. Invaginations of the inner membrane complex were seen in the gametocytes and may be concerned with nutrition. The meronts were irregularly shaped before division, a feature unusual among members of the Apicomplexa. The unusual presence of a parasitic protozoan within neutrophils, in some way interfering with or modulating the microbicidal activity of such cells, is discussed

AgNP-PVP-meglumine antimoniate nanocomposite reduces Leishmania amazonensis infection in macrophages

Institute Evandro Chagas (IEC), Department of Health Surveillance, Ministry of Health: Laboratory of Electron Microscopy and Laboratory of Superficial and Systemic Mycoses. Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), grant number: 424101/2016-5. Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - finance code 001.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Microscopia Eletrônica. Belém, PA, Brasil / Federal University of Pará. Postgraduate Program in Biology of Infectious and Parasitic Agents. Belém, PA, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Microscopia Eletrônica. Belém, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Microscopia Eletrônica. Belém, PA, Brasil / Federal University of Pará. Postgraduate Program in Biology of Infectious and Parasitic Agents. Belém, PA, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Microscopia Eletrônica. Belém, PA, Brasil.Background: Leishmaniasis is an infectious disease caused by parasites of the genus Leishmania and presents different clinical manifestations. The adverse effects, immunosuppression and resistant strains associated with this disease necessitate the development of new drugs. Nanoparticles have shown potential as alternative antileishmanial drugs. We showed in a previous study the biosynthesis, characterization and ideal concentration of a nanocomposite that promoted leishmanicidal activity. In the present study, we conducted a specific analysis to show the mechanism of action of AgNP-PVP-MA (silver nanoparticle-polyvinylpyrrolidone-[meglumine antimoniate (Glucantime (R))]) nanocomposite during Leishmania amazonensis infection in vitro. Results: Through ultrastructural analysis, we observed significant alterations, such as the presence of small vesicles in the flagellar pocket and in the extracellular membrane, myelin-like structure formation in the Golgi complex and mitochondria, flagellum and plasma membrane rupture, and electrodense material deposition at the edges of the parasite nucleus in both evolutive forms. Furthermore, the Leishmania parasite infection index in macrophages decreased significantly after treatment, and nitric oxide and reactive oxygen species production levels were determined. Additionally, inflammatory, and pro-inflammatory cytokine and chemokine production levels were evaluated. The IL-4, TNF-alpha and MIP-1 alpha levels increased significantly, while the IL-17 A level decreased significantly after treatment. Conclusions: Thus, we demonstrate in this study that the AgNP-PVP-MA nanocomposite has leishmanial potential, and the mechanism of action was demonstrated for the first time, showing that this bioproduct seems to be a potential alternative treatment for leishmaniasis

Cytochemical and functional characterization of blood and inflammatory cells from the lizard Ameiva ameiva

Conselho Nacional de Desenvolvimento Científico e Tecnológico (MCT-CNPq), Fundação de Coordenação de Pessoal de Nível Superior (CAPES), Fundação Carlos Chagas Filho de Amparo a Pesquisa do Estado do Rio de Janeiro (FAPERJ), Fundação Estadual do Norte Fluminense (FENORTE), Financiadora de Estudos e Projetos (FINEP), Programa Nacional de Cooperação Acadêmica (PROCAD) and Programa de Núcleos de Excelência (PRONEX)Universidade Estadual do Pará. Departamento de Fisiologia. Belém, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Microscopia Eletrônica. Belém, PA, Brasil.Universidade Federal do Pará. Centro de Ciências Biológicas. Laboratório de Parasitologia. Belém, PA, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Biofísica Carlos Chagas Filho. Laboratório de Ultraestrutura Celular Hertha Meyer. Rio de Janeiro, RJ, Brasil.Universidade Estadual do Norte Fluminense. Centro de Biociências e Biotecnologia. Laboratório de Biologia Celular e Tecidual. Campos de Goytacazes, RJ, Brasil.The fine structure and differential cell count of blood and coelomic exudate leukocytes were studied with the aim to identify granulocytes from Ameiva ameiva, a lizard distributed in the tropical regions of the Americas. Blood leukocytes were separated with a Percoll cushion and coelomic exudate cells were obtained 24 h after intracoelomic thioglycollate injection. In the blood, erythrocytes, monocytes, thrombocytes, lymphocytes, plasma cells and four types of granulocytes were identified based on their morphology and cytochemistry. Types I and III granulocytes had round intracytoplasmic granules with the same basic morphology; however, type III granulocyte had a bilobued nucleus and higher amounts of heterochromatin suggesting an advance stage of maturation. Type II granulocytes had fusiformic granules and more mitochondria. Type IV granulocytes were classified as the basophil mammalian counterpart based on their morphology and relative number. Macrophages and granulocytes type III were found in the normal coelomic cavity. However, after the thioglycollate injection the number of type III granulocyte increased. Granulocytes found in the coelomic cavity were related to type III blood granulocyte based on the morphology and cytochemical localization of alkaline phosphatase and basic proteins in their intracytoplasmic granules. Differential blood leukocyte counts showed a predominance of type III granulocyte followed by lymphocyte, type I granulocyte, type II granulocyte, monocyte and type IV granulocyte. Taken together, these results indicate that types I and III granulocytes correspond to the mammalian neutrophils/heterophils and type II to the eosinophil granulocytes

Early and late neuropathological features of meningoencephalitis associated with Maraba virus infection

Universidade Estadual do Pará, Instituto Evandro Chagas, and Universidade Federal do Pará.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Microscopia Eletrônica. Ananindeua, PA, Brasil.Universidade Federal do Pará. Hospital Universitário João Barros Barreto. Instituto de Ciências Biológicas. Laboratório de Neurodegeneração e Infecção. Belém, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Microscopia Eletrônica. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Microscopia Eletrônica. Ananindeua, PA, Brasil / Universidade Federal do Pará. Hospital Universitário João Barros Barreto. Instituto de Ciências Biológicas. Laboratório de Neurodegeneração e Infecção. Belém, PA, Brasil.Universidade Federal do Pará. Núcleo de Medicina Tropical. Belém, PA, Brasil.Universidade Federal do Pará. Hospital Universitário João Barros Barreto. Instituto de Ciências Biológicas. Laboratório de Neurodegeneração e Infecção. Belém, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Microscopia Eletrônica. Ananindeua, PA, Brasil / Universidade Federal do Pará. Hospital Universitário João Barros Barreto. Instituto de Ciências Biológicas. Laboratório de Neurodegeneração e Infecção. Belém, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Microscopia Eletrônica. Ananindeua, PA, Brasil.Maraba virus is a member of the genus Vesiculovirus of the Rhabdoviridae family that was isolated in 1983 from sandflies captured in the municipality of Maraba, state of Pará, Amazônia, Brazil. Despite 30 years having passed since its isolation, little is known about the neuropathology induced by the Maraba virus. Accordingly, in this study the histopathological features, inflammatory glial changes, cytokine concentrations, and nitric oxide activity in the encephalon of adult mice subjected to Maraba virus nostril infection were evaluated. The results showed that 6 days after intranasal inoculation, severe neuropathological-associated disease signs appeared, including edema, necrosis and pyknosis of neurons, generalized congestion of encephalic vessels, and intra- and perivascular meningeal lymphocytic infiltrates in several brain regions. Immunolabeling of viral antigens was observed in almost all central nervous system (CNS) areas and this was associated with intense microglial activation and astrogliosis. Compared to control animals, infected mice showed significant increases in interleukin (IL)-6, tumor necrosis factor (TNF)-a, interferon (INF)-g, MCP-1, nitric oxide, and encephalic cytokine levels. We suggest that an exacerbated inflammatory response in several regions of the CNS of adult BALB/c mice might be responsible for their deaths

Phagocytosis of Fonsecae pedrosoi conidia, but not sclerotic cells caused by Langerhans cells, inhibits CD40 and B7-2 expression

Universidade do Estado do Pará. Laboratório de Dermato-Imunologia. Belém, PA, Brazil / Unidade de Referência em Dermatologia Sanitária do Estado do Pará Dr Marcello Candia. Marituba, PA, Brazil / Universidade Federal do Pará. Departamento de Farmácia. Belém, PA, Brazil / Universidade Federal do Rio de Janeiro. Laboratório de Biologia Celular de Fungos. Rio de Janeiro, RJ, Brazil.Universidade do Estado do Pará. Laboratório de Dermato-Imunologia. Belém, PA, Brazil.Universidade do Estado do Pará. Laboratório de Dermato-Imunologia. Belém, PA, Brazil / Universidade do Estado do Pará. Serviço de Dermatologia. Belém, PA, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Microscopia Eletrônica. Ananindeua, PA, Brasil.Universidade Federal do Rio de Janeiro. Laboratório de Biologia Celular de Fungos. Rio de Janeiro, RJ, Brazil.Universidade do Estado do Pará. Laboratório de Dermato-Imunologia. Belém, PA, Brazil / Universidade Federal do Pará. Departamento de Patologia. Belém, PA, Brazil.Fonsecaea pedrosoi is the major etiological agent of chromoblastomycosis, a chronic, suppurative, granulomatous mycosis usually confined to skin and subcutaneous tissues, presenting a worldwide distribution. The host defense mechanisms in chromoblastomycosis have not been extensively investigated. Langerhans cells (LC) are bone-marrow-derived, dendritic antigen-presenting cells of the epidermis, which constitutively express major histocompatibility complex (MHC) class II, and comprise 1–3% of total epidermal cells. LC are localized in suprabasal layers of the epidermis and in mucosa, where they play important roles in skin immune responses. The purpose of the present study was to evaluate the interaction of F. pedrosoi conidia or sclerotic cells with LC purified from BALB/c mice skin. We demonstrate here that LC phagocytose F. pedrosoi conidia but not sclerotic cells in the first 3 h of interaction, inhibiting hyphae formation during 12- hour coculture from both forms, internalized or not. Also, LC maturation, analyzed using CD40 and B7-2 expression, was inhibited by conidia, but not by sclerotic cells, indicating an important innate immunity function of LC against F. pedrosoi infection in these mice