RESUMO
AIMS: Chagas disease is a neglected tropical disease. The ability of Trypanosoma cruzi to survive within phagocytes is likely a critical factor for T. cruzi dissemination in the host. For control of the parasite load and host survival, macrophage action is required. Concanavalin-A (Con-A) presents properties that modulate immune functions and protect hosts from several experimental infectious diseases. Here, we evaluated the effects of Con-A on peritoneal macrophages as well as on the course of experimental infection by T. cruzi. MAIN METHODS: BALB/c mice, a susceptible model for T. cruzi infection, were treated with Con-A via the intraperitoneal route and 3 days later infected with T. cruzi. We quantified parasitemia, cytokines and nitric oxide (NO). Peritoneal exudate and macrophages were collected for macrophage phenotyping and cell viability, NO and cytokine detection, as well as for T. cruzi internalization and release index determination. KEY FINDINGS: Con-A treatment induced IL-17a and NO production by cells from the peritoneal cavity, and M1 marker expression predominated on peritoneal macrophages. These cells are also more prone to producing TNF-α, IL-6 and NO when infected by T. cruzi and show high trypanocidal capacity. Due to a hostile peritoneal microenvironment caused by Con-A, which induces macrophage cNOS and iNOS expression, infected BALB/c mice showed reduced parasitemia and an increased survival rate. SIGNIFICANCE: We conclude that Con-A can induce peritoneal M1 macrophage polarization to increase trypanocidal activity, resulting in ameliorated systemic infection in a susceptible experimental model.
Assuntos
Polaridade Celular , Doença de Chagas/patologia , Concanavalina A/farmacologia , Interleucina-17/metabolismo , Macrófagos Peritoneais/patologia , Macrófagos Peritoneais/parasitologia , Óxido Nítrico/metabolismo , Trypanosoma cruzi/fisiologia , Animais , Polaridade Celular/efeitos dos fármacos , Doença de Chagas/metabolismo , Feminino , Macrófagos Peritoneais/efeitos dos fármacos , Camundongos Endogâmicos BALB C , Óxido Nítrico Sintase Tipo II/metabolismo , Parasitemia/metabolismo , Parasitemia/patologia , Trypanosoma cruzi/efeitos dos fármacosRESUMO
We evaluated the influence of metabolic syndrome (MS) on acute Trypanosoma cruzi infection. Obese Swiss mice, 70 days of age, were subjected to intraperitoneal infection with 5 × 102 trypomastigotes of the Y strain. Cardiovascular, oxidative, inflammatory, and metabolic parameters were evaluated in infected and non-infected mice. We observed higher parasitaemia in the infected obese group (IOG) than in the infected control group (ICG) 13 and 15 days post-infection. All IOG animals died by 19 days post-infection (dpi), whereas 87.5% of the ICG survived to 30 days. Increased plasma nitrite levels in adipose tissue and the aorta were observed in the IOG. Higher INF-γ and MCP-1 concentrations and lower IL-10 concentrations were observed in the IOG compared to those in the ICG. Decreased insulin sensitivity was observed in obese animals, which was accentuated after infection. Higher parasitic loads were found in adipose and hepatic tissue, and increases in oxidative stress in cardiac, hepatic, and adipose tissues were characteristics of the IOG group. Thus, MS exacerbates experimental Chagas disease, resulting in greater damage and decreased survival in infected animals, and might be a warning sign that MS can influence other pathologies.
Assuntos
Tecido Adiposo/metabolismo , Doença de Chagas/metabolismo , Inflamação/metabolismo , Fígado/metabolismo , Síndrome Metabólica/metabolismo , Miocárdio/metabolismo , Estresse Oxidativo/fisiologia , Tecido Adiposo/patologia , Animais , Doença de Chagas/complicações , Doença de Chagas/patologia , Citocinas/sangue , Modelos Animais de Doenças , Fígado Gorduroso/metabolismo , Fígado Gorduroso/patologia , Inflamação/complicações , Inflamação/patologia , Resistência à Insulina/fisiologia , Fígado/patologia , Masculino , Síndrome Metabólica/complicações , Síndrome Metabólica/patologia , Camundongos , Miocárdio/patologia , Trypanosoma cruziRESUMO
Infection with the protozoan Trypanosoma cruzi causes Chagas disease and consequently leads to severe inflammatory heart condition; however, the mechanisms driving this inflammatory response have not been completely elucidated. Nitric oxide (NO) is a key mediator of parasite killing in T. cruzi-infected mice, and previous studies have suggested that leukotrienes (LTs) essentially regulate the NO activity in the heart. We used infected 5-lipoxygenase-deficient mice (5-LO-/-) to explore the participation of nitric oxide synthase isoforms, inducible (iNOS) and constitutive (cNOS), in heart injury, cytokine profile, and oxidative stress during the early stage of T. cruzi infection. Our evidence suggests that the cNOS of the host is involved in the resistance of 5-LO-/- mice during T. cruzi infection. iNOS inhibition generated a remarkable increase in T. cruzi infection in the blood and heart of mice, whereas cNOS inhibition reduced cardiac parasitism (amastigote nests). Furthermore, this inhibition associates with a higher IFN-γ production and lower lipid peroxidation status. These data provide a better understanding about the influence of NO-interfering therapies for the inflammatory response toward T. cruzi infection.
Assuntos
Araquidonato 5-Lipoxigenase/sangue , Doença de Chagas/sangue , Doença de Chagas/enzimologia , Animais , Antioxidantes/metabolismo , Citocinas/sangue , Camundongos , Camundongos Knockout , Óxido Nítrico/sangue , Óxido Nítrico Sintase Tipo II/sangue , Trypanosoma cruzi/patogenicidadeRESUMO
During the onset of Trypanosoma cruzi infection, an effective immune response is necessary to control parasite replication and ensure host survival. Macrophages have a central role in innate immunity, acting as an important trypanocidal cell and triggering the adaptive immune response through antigen presentation and cytokine production. However, T. cruzi displays immune evasion mechanisms that allow infection and replication in macrophages, favoring its chronic persistence. One potential mechanism is the release of T. cruzi strain Y extracellular vesicle (EV Y), which participate in intracellular communication by carrying functional molecules that signal host cells and can modulate the immune response. The present work aimed to evaluate immune modulation by EV Y in C57BL/6 mice, a prototype resistant to infection by T. cruzi strain Y, and the effects of direct EV Y stimulation of macrophages in vitro. EV Y inoculation in mice prior to T. cruzi infection resulted in increased parasitemia, elevated cardiac parasitism, decreased plasma nitric oxide (NO), reduced NO production by spleen cells, and modulation of cytokine production, with a reduction in TNF-α in plasma and decreased production of TNF-α and IL-6 by spleen cells from infected animals. In vitro assays using bone marrow-derived macrophages showed that stimulation with EV Y prior to infection by T. cruzi increased the parasite internalization rate and release of infective trypomastigotes by these cells. In this same scenario, EV Y induced lipid body formation and prostaglandin E2 (PGE2) production by macrophages even in the absence of T. cruzi. In infected macrophages, EV Y decreased production of PGE2 and cytokines TNF-α and IL-6 24 h after infection. These results suggest that EV Y modulates the host response in favor of the parasite and indicates a role for lipid bodies and PGE2 in immune modulation exerted by EVs.
Assuntos
Doença de Chagas/imunologia , Vesículas Extracelulares/imunologia , Interações Hospedeiro-Parasita/imunologia , Macrófagos Peritoneais/imunologia , Trypanosoma cruzi/imunologia , Animais , Doença de Chagas/parasitologia , Chlorocebus aethiops , Dinoprostona/imunologia , Dinoprostona/metabolismo , Modelos Animais de Doenças , Humanos , Evasão da Resposta Imune , Gotículas Lipídicas/imunologia , Gotículas Lipídicas/metabolismo , Ativação de Macrófagos/imunologia , Macrófagos Peritoneais/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Baço/citologia , Baço/imunologia , Trypanosoma cruzi/metabolismo , Células VeroRESUMO
American Tegumentar Leishmaniasis (ATL) is an infectious disease caused by Leishmania parasites with ineffective treatment. The properties of propolis have been studied in different experimental studies, however, few works have investigated the effects of propolis on human-derived peripheral blood mononuclear cells (PBMC) in leishmaniasis models. Thus, we investigate the immunomodulatory effects of propolis treatment on PBMC from ATL patients and on PBMC from healthy donors infected with Leishmania braziliensis. Our data demonstrate that propolis pretreatment shows immunomodulatory effects on both healthy donors and ATL patients adherent cells, increasing IL-4 and IL-17 and decreasing IL-10, in either the presence or absence of the L. braziliensis infection, demonstrating that propolis contributes with the decrease of the inflammation and could also contribute with parasite control.
Assuntos
Anti-Inflamatórios/uso terapêutico , Leishmania braziliensis/imunologia , Leishmaniose/imunologia , Leucócitos Mononucleares/efeitos dos fármacos , Própole/uso terapêutico , Pele/patologia , Adulto , Idoso , Brasil , Citocinas/metabolismo , Feminino , Humanos , Imunomodulação , Leucócitos Mononucleares/fisiologia , Masculino , Pessoa de Meia-Idade , Óxido Nítrico/metabolismo , Pele/parasitologiaRESUMO
O objetivo deste trabalho foi determinar a presença de parasitas intestinais em amostras de alfaces e almeirões comercializadas em feira livre e supermercado da cidade de Londrina, Paraná. Foram coletadas e analisadas oito amostras de alface (Lactuca sativa) e oito amostras de almeirão (Cichirium intybus). A análise foi realizada no Laboratório de Extensão e Pesquisas em Enteroparasitoses L.E.P.En. Foram encontrados cistos de Endolimax nana, Entamoeba coli, Entamoeba histolytica, Balantidium coli, larvas de Strongyloides stercoralis, larvas e ovos de Ancilostomídeos e ovos de Ascaris spp. Esses resultados demonstram a necessidade de medidas higiênico-sanitárias, visando à educação da comunidade, possibilitando a melhoria na condição de vida da população.
The objective of this study was to determine the presence of intestinal parasites in samples of lettuce andchicory sold in street market and supermarket Londrina city, Paraná. For this purpose, eight samples of lettuce (Lactuca sativa) and eight samples of chicory (Cichorium intybus) were collected and analyzed. The analysis was performed at the Laboratory for Extension and Research in Enteroparasitosis L.E.P.En. Cysts of Endolimax nana, Entamoeba coli, Entamoeba histolytica, Balantidium coli and Strongyloides stercoralis larvae, Hookworm larvae and eggs, and eggs of Ascaris spp were found in the samples. These results demonstrate the need for hygienic-sanitary measures, aimed at educating the community, enabling the improvement in living conditions of the population.
Assuntos
Doenças Parasitárias , Helmintíase , Pesquisa , Saúde Pública , Verduras/parasitologiaRESUMO
Trypanosoma cruzi is the causative agent of Chagas disease. Approximately 8 million people are thought to be affected with this disease worldwide. T. cruzi infection causes an intense inflammatory response, which is critical for the control of parasite proliferation and disease development. Nitric oxide-donating nonsteroidal anti-inflammatory drugs (NO-NSAIDs) are an emergent class of pharmaceutical derivatives with promising utility as chemopreventive agents. In this study, we investigated the effect of NO-indomethacin on parasite burden, cell invasion, and oxidative stress in erythrocytes during the acute phase of infection. NO-indomethacin was dissolved in dimethyl formamide followed by i.p. administration of 50 ppm into mice 30 min after infection with 5×10(3) blood trypomastigote forms (Y strain). The drug was administered every day until the animals died. Control animals received 100 µL of drug vehicle via the same route. Within the NO-indomethacin-treatment group, parasitemia and mortality (100%) were higher and oxidative stress in erythrocytes, anemia, and entry of parasites into macrophages were significantly greater than that seen in controls. Increase in the entry and survival of intracellular T. cruzi was associated with inhibition of nitric oxide production by macrophages treated with NO-indomethacin (2.5 µM). The results of this study provide strong evidence that NO-NSAIDs potently inhibit nitric oxide production, suggesting that NO-NSAID-based therapies against infections would be difficult to design and would require caution.
Assuntos
Indometacina/análogos & derivados , Nitratos/farmacologia , Óxido Nítrico/metabolismo , Estresse Oxidativo , Trypanosoma cruzi/patogenicidade , Anemia/metabolismo , Anemia/patologia , Animais , Células Cultivadas , Suscetibilidade a Doenças , Eritrócitos/metabolismo , Feminino , Indometacina/química , Indometacina/farmacologia , Macrófagos/citologia , Macrófagos/parasitologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Nitratos/química , Estresse Oxidativo/efeitos dos fármacos , Parasitemia/tratamento farmacológico , Parasitemia/mortalidade , Parasitemia/patologiaRESUMO
The intracellular protozoan parasite Trypanosoma cruzi causes Chagas disease, a serious disorder that affects millions of people in Latin America. Cell invasion by T. cruzi and its intracellular replication are essential to the parasite's life cycle and for the development of Chagas disease. Here, we present evidence suggesting the involvement of the host's cyclooxygenase (COX) enzyme during T. cruzi invasion. Pharmacological antagonist for COX-1, aspirin (ASA), caused marked inhibition of T. cruzi infection when peritoneal macrophages were pretreated with ASA for 30 min at 37°C before inoculation. This inhibition was associated with increased production of IL-1ß and nitric oxide (NO(â)) by macrophages. The treatment of macrophages with either NOS inhibitors or prostaglandin E2 (PGE2) restored the invasive action of T. cruzi in macrophages previously treated with ASA. Lipoxin ALX-receptor antagonist Boc2 reversed the inhibitory effect of ASA on trypomastigote invasion. Our results indicate that PGE2, NO(â), and lipoxins are involved in the regulation of anti-T. cruzi activity by macrophages, providing a better understanding of the role of prostaglandins in innate inflammatory response to T. cruzi infection as well as adding a new perspective to specific immune interventions.
Assuntos
Aspirina/farmacologia , Macrófagos Peritoneais/parasitologia , Trypanosoma cruzi/patogenicidade , Animais , Células Cultivadas , Ensaio de Imunoadsorção Enzimática , Feminino , Imuno-Histoquímica , Interleucina-1beta/metabolismo , Macrófagos Peritoneais/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Óxido Nítrico Sintase Tipo II , Trypanosoma cruzi/efeitos dos fármacosRESUMO
Trypanosoma cruzi, the causative agent of Chagas' disease (CD), is a substantial public health concern in Latin America. Laboratory mice inoculated with T. cruzi have served as important animal models of acute CD. Host hypoferremic responses occur during T. cruzi infection; therefore, it has been hypothesized that T. cruzi requires iron for optimal growth in host cells and, unlike extracellular pathogens, may benefit from host hypoferremic responses. Recent technological improvements of X-ray fluorescence are useful for diagnostics or monitoring in biomedical applications. The goal of our study was to determine whether the iron availabilities in Swiss and C57BL/6 mice differ during the acute phase of T. cruzi infection and whether the availability correlates with oxidative stress in the susceptible and resistant phenotypes identified in these mice. Our results showed that the decrease in iron levels in the skin of resistant infected mice correlated with the increase in oxidative stress associated with anemia and the reduction in parasite burden.
Assuntos
Doença de Chagas/metabolismo , Deficiências de Ferro , Ferro/metabolismo , Pele/metabolismo , Trypanosoma cruzi/metabolismo , Anemia/sangue , Anemia/metabolismo , Anemia/parasitologia , Animais , Doença de Chagas/sangue , Doença de Chagas/parasitologia , Eritrócitos/metabolismo , Fluorescência , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Estresse Oxidativo/fisiologia , Saúde Pública , Raios XRESUMO
Leukotrienes are important mediators of inflammatory responses. In this study, we investigated the effect of the absence of 5-lipoxygenase (5-LO)-derived leukotrienes on levels of cytokines, nitric oxide (NO) and iNOS expression in cardiac tissue of mice infected with Trypanosoma cruzi, the agent of Chagas' disease. NO is a key mediator of parasite killing in mice experimentally infected with T. cruzi, and previous studies have suggested that leukotrienes, such as LTB(4), induces NO synthesis in T. cruzi-infected macrophages and plays a relevant role in the killing of parasite in a NO-dependent manner. We therefore investigated whether leukotrienes would have a similar role in vivo in controlling the parasite burden by regulating NO activity. We have made the striking observation that absence of 5-LO-derived leukotrienes results in increased NO and IL-6 production in the plasma with a concomitant decrease in the expression of iNOS in the cardiac tissue on day 12 after T. cruzi infection. These findings indicate that endogenous leukotrienes are important regulators of NO activity in the heart and therefore influence the cardiac parasite burden without exerting a direct action on IL-6 production in the acute phase of infection with T. cruzi.
Assuntos
Araquidonato 5-Lipoxigenase/metabolismo , Cardiomiopatia Chagásica/metabolismo , Citocinas/metabolismo , Leucotrienos/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Óxido Nítrico/metabolismo , Doença Aguda , Animais , Araquidonato 5-Lipoxigenase/genética , Citocinas/sangue , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Coração/parasitologia , Imuno-Histoquímica , Masculino , Camundongos , Camundongos Knockout , Miocárdio/metabolismo , Óxido Nítrico/sangueRESUMO
Chagas' disease is accompanied by severe anemia and oxidative stress, which may contribute to mortality. In this study, we investigated the role of 5-lipoxygenase (5-LO) in the control of parasitism and anemia associated with oxidative damage of erythrocytes in experimental Trypanosoma cruzi infection. Wild-type C57BL/6, 129Sv mice treated or not with nordihydroguaiaretic acid (NDGA, 5-LO inhibitor), mice lacking the 5-LO enzyme gene (5-LO(-/-)) and inducible nitric oxide synthase gene (iNOS(-/-)) were infected with the Y strain of T. cruzi. Impairment of 5-LO resulted in increased numbers of trypomastigote forms in the blood and amastigote forms in the heart of infected mice. We assessed oxidative stress in erythrocytes by measuring oxygen uptake, induction time and chemiluminescence following treatment with tert-butyl hydroperoxide (TBH). Our results show that 5-LO metabolites increased lipid peroxidation levels in erythrocytes during the early phase of murine T. cruzi infection. NDGA treatment reduced oxidative damage of erythrocytes in C57BL/6 T. cruzi-infected mice but not in C57BL/6 iNOS(-/-) infected mice, showing that the action of NDGA is dependent on endogenous nitric oxide (NO). In addition, our results show that 5-LO metabolites do not participate directly in the development of anemia in infected mice. We conclude that 5-LO products may not only play a major role in controlling heart tissue parasitism, i.e., host resistance to acute infection with T. cruziin vivo, but in the event of an infection also play an important part in erythrocyte oxidative stress, an NO-dependent effect.
Assuntos
Araquidonato 5-Lipoxigenase/metabolismo , Doença de Chagas/imunologia , Eritrócitos/enzimologia , Óxido Nítrico Sintase Tipo II/metabolismo , Trypanosoma cruzi/imunologia , Animais , Araquidonato 5-Lipoxigenase/genética , Doença de Chagas/enzimologia , Doença de Chagas/parasitologia , Modelos Animais de Doenças , Eritrócitos/imunologia , Eritrócitos/parasitologia , Inibidores de Lipoxigenase/farmacologia , Masculino , Masoprocol/farmacologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Óxido Nítrico/imunologia , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo II/imunologia , Estresse Oxidativo/imunologiaRESUMO
Investigação da associação do polimorfismo do receptor de quimiocinas CCR5 com câncer cervical em pacientes HPV positivos. O HPV é um dos maiores responsáveis pelo desenvolvimento do câncer cervical. É conhecido que asquimiocinas são importantes determinantes da resposta inflamatória precoce. O produto do gene do receptor de quimiocinas CCR5 está envolvido na quimiotaxia de leucócitos para sítios inflamatórios. No presente estudo, reações em cadeia da polimerase (PCR) de amostras de DNA genômico, utilizando iniciadores específicos para CCR5 que flanqueam a região de deleção, foram utilizadas para detectar produto de 225 bp para o alelo normal e193 bp para o alelo que apresenta a deleção de 32 bp. O genótipo selvagem foi o mais prevalente em ambos os grupos e não houve diferença estatisticamente significativa, com χ2= 1,519 (2 graus de liberdade; p > 0,05). Uma vez que a prevalência de indivíduos portadores do alelo D32 é pequena, são necessários mais estudos a fim de elucidar o papel do CCR5 no cancer cervical.
HPV is one of the most frequent causes for the development of cervical cancer. It is known that chemokines are important determinants of early inflammatory responses. The CC chemokine receptor 5 (CCR5) gene is involved in the chemotaxis of leukocytes toward inflammation sites. In the present study, polymerase chain reactions (PCR) in genomic DNA samples, using specific CCR5 oligonucleotide primers surrounding the breakpoint deletion, detected a 225 bp product from the normal CCR5allele and a 193 bp product from the 32 bp deletion allele. The wild type genotype was prevalent in both group, but it was not statistically significant, with χ2 = 1.519 (2 degrees of freedom; p > 0.05). As there are a small number of D32 allele carriers, further studies areneeded to clarify the role of CCR5 in the cervical cancer.