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Rhizophora mangle is commonly used in traditional medicine to treat infections, reduce inflammation, and promote healing. This study aimed to analyze the phytochemical profile of the methanolic extract of R. mangle leaves (MELRm) and evaluate its in vitro schistosomicidal activity against Schistosoma mansoni as well as its cytotoxicity. Plant material was collected in Itamaracá City, Pernambuco, Brazil. The extract was analyzed using UV/Vis spectrophotometry and high-performance liquid chromatography (HPLC). The motility, mortality, and cell viability of adult worms were assessed in a schistosomicidal assay, while cytotoxicity was evaluated through a colorimetric assay with MTT on RAW 264.7 cells. The primary compounds identified in MELRm were phenolic compounds. In the schistosomicidal assay, all concentrations of MELRs induced changes in the motility of adult worms. At a concentration of 400 µg/mL, MELRs resulted in 56.25% mortality after 72 h of incubation. After 120 h, mortality rates of 75%, 62.5%, and 50% were observed at MELRm concentrations of 400, 200, and 100 µg/mL, respectively. No eggs were detected at any MELRm concentration. MELRs did not show cytotoxicity towards RAW 264.7 cells at the concentrations tested. These results indicate that MELRs demonstrate schistosomicidal activity in vitro, suggesting they are promising candidates for in vivo studies.
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The objective of the study was to evaluate the in vitro and in vivo schistosomicidal activity of sanguinarine (SA) on Schistosoma mansoni and its in silico pharmacokinetic parameters. ADMET parameters and oral bioavailability were evaluated using the PkCSM and SwissADME platforms, respectively. The activity of SA in vitro, at the concentrations of 1.0-25 µM, was analyzed through the parameters of motility, mortality, and cell viability of the worms at intervals of 3-24 h. Mice were infected with cercariae and treated by gavage with SA (5 mg/kg/day, in a single dose or two doses of 2.5 mg/kg every 12 h for 5 consecutive days) on the 1st (skin schistosomula), 14th (pulmonary schistosomula), 28th (young worms), and 45th (adult worms) days after infection. In vitro and in vivo praziquantel was the control. In vitro, SA showed schistosomicidal activity against schistosomula, young worms, and couples; with total mortality and reduced cell viability at low concentrations and incubation time. In a single dose of 5 mg/kg/day, SA reduces the total worm load by 47.6%, 54%, 55.2%, and 27.1%, and female worms at 52.0%, 39.1%, 52.7%, and 20.2%, respectively, results which are similar to the 2.5 mg/kg/day dose. SA reduced the load of eggs in the liver, and in histopathological and histomorphometric analyses, there was a reduction in the number and volume of hepatic granulomas, which exhibited less inflammatory infiltrate. SA has promising in vitro and in vivo schistosomicidal activity against different developmental stages of S. mansoni, in addition to reducing granulomatous liver lesions. Furthermore, in silico, SA showed good predictive pharmacokinetic ADMET profiles.
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Alcaloides , Antiinfecciosos , Isoquinolinas , Esquistosomicidas , Femenino , Animales , Ratones , Antiparasitarios , Schistosoma mansoni , Benzofenantridinas/farmacología , Alcaloides/farmacologíaRESUMEN
Anti-Ascaris lumbricoides (Asc) IgE and IgG can immunomodulate the allergy; however, the influence of these isotypes has not been investigated in the giardiasis and allergy. Therefore, the frequency of respiratory allergy (RA) symptoms in Giardia lamblia-infected children, with or without anti-Asc IgE, IgG1, or IgG4 and Th1, Th2/Treg, and Th17 cytokine production, was evaluated. We performed a case-control study with children aged 2-10 years old selected by questionnaire and stool exams to form the groups: infected or uninfected with RA (G-RA, n = 55; nG-RA, n = 43); infected and uninfected without RA (G-nRA, n = 59; nG-nRA, n = 54). We performed blood leukocyte counts and in vitro culture. Cytokine levels in the supernatants (CBA), serum total IgE and anti-Asc IgE (ImmunoCAP), IgG1, IgG4, and total IgA (ELISA) were measured. Infection was not associated with allergy. Infected children showed increased levels of anti-Asc IgG1, IL-2, IFN-γ, IL-4, and IL-10. There was a lower frequency of allergy-related symptoms in anti-Asc IgG1-positive children than IgG1-negative (OR = 0.38; CI = 0.17-0.90, p = 0.027) and few eosinophils in G-RA than in G-nRA and more in G-nRA than in nG-nRA, whereas TNF-α levels were higher in the G-RA than in the nG-nRA group. For infected and positive anti-Asc IgG1, there was higher TNF-α and IL-10 production than G/-IgG1. IL-10 levels were lower in nG/ + IgG1 than in infected or non-infected, and both were negative for anti-Asc IgG1. Th1/Th2/IL-10 profiles were stimulated in the infected patients, and in those with circulating anti-Asc IgG1, the TNF-α production was strengthened with a lower risk for respiratory allergy symptoms.
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Giardia lamblia , Hipersensibilidad , Animales , Humanos , Niño , Preescolar , Interleucina-10 , Ascaris , Factor de Necrosis Tumoral alfa , Estudios de Casos y Controles , Hipersensibilidad/complicaciones , Citocinas , Inmunoglobulina G , Inmunoglobulina ERESUMEN
Schistosomiasis, a potentially fatal chronic disease whose etiological agents are blood trematode worms of the genus Schistosoma spp., is one of the most prevalent and debilitating neglected diseases. The treatment of schistosomiasis depends exclusively on praziquantel (PZQ), a drug that has been used since the 1970s and that already has reports of reduced therapeutic efficacy, related with the development of Schistosoma-resistant or -tolerant strains. Therefore, the search for new therapeutic alternatives is an urgent need. Plumbagin (PLUM), a naphthoquinone isolated from the roots of plants of the genus Plumbago, has aroused interest in research due to its antiparasitic properties against protozoa and helminths. Here, we evaluated the in vivo schistosomicidal potential of PLUM against Schistosoma mansoni and the in silico pharmacokinetic parameters. ADMET parameters and oral bioavailability were evaluated using the PkCSM and SwissADME platforms, respectively. The study was carried out with five groups of infected mice and divided as follows: an untreated control group, a control group treated with PZQ, and three groups treated orally with 8, 16, or 32 mg/kg of PLUM. After treatment, the Kato-Katz technique was performed to evaluate a quantity of eggs in the feces (EPG). The animals were euthanized for worm recovery, intestine samples were collected to evaluate the oviposition pattern, the load of eggs was determined on the hepatic and intestinal tissues and for the histopathological and histomorphometric evaluation of tissue and hepatic granulomas. PLUM reduced EPG by 65.27, 70.52, and 82.49%, reduced the total worm load by 46.7, 55.25, and 72.4%, and the female worm load by 44.01, 52.76, and 71.16%, for doses of 8, 16, and 32 mg/kg, respectively. PLUM also significantly reduced the number of immature eggs and increased the number of dead eggs in the oogram. A reduction of 36.11, 46.46, and 64.14% in eggs in the hepatic tissue, and 57.22, 65.18, and 80.5% in the intestinal tissue were also observed at doses of 8, 16, and 32 mg/kg, respectively. At all doses, PLUM demonstrated an effect on the histopathological and histomorphometric parameters of the hepatic granuloma, with a reduction of 41.11, 48.47, and 70.55% in the numerical density of the granulomas and 49.56, 57.63, and 71.21% in the volume, respectively. PLUM presented itself as a promising in vivo antiparasitic candidate against S. mansoni, acting not only on parasitological parameters but also on hepatic granuloma. Furthermore, in silico, PLUM showed good predictive pharmacokinetic profiles by ADMET.
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Schistosoma mansoni infections, particularly egg antigens, induce Th2-dominant granulomatous responses accompanied by remarkable immunoregulatory mechanisms that avoid intense fibrosis. Interleukin (IL)-33 is a cytokine that stimulates the early activation of Th2 responses, and its soluble ST2 receptor (sST2) avoids granulomatous response, as well as CXCL9 and CXCL10 chemokines that have antifibrotic activity. However, in schistosomiasis, these molecules have not been suitably studied. Therefore, this study aimed to measure IL-33 and sST2 RNA, cytokines, and chemokines in peripheral blood cultures from individuals living in schistosomiasis-endemic areas. Peripheral blood cells from individuals with S. mansoni (n = 34) and non-infected individuals (n = 31) were cultured under mitogen stimulation. Supernatant chemokines and cytokines were evaluated using a cytometric bead array, and IL-33 and sST2 mRNA expression was measured using qPCR. Infected individuals showed higher levels of CXCL8, CXCL9, CXCL10, IFN-γ, TNF-α, IL-6, IL-2, IL-4, and IL-10; there was a lower expression of IL-33 mRNA and similar expression of sST2mRNA in infected than non-infected individuals. In conclusion, for the first time, we demonstrated lower IL-33mRNA expression and high levels of the antifibrotic chemokines CXCL9 and CXCL10 in schistosomiasis mansoni, which could control exacerbations of the disease in individuals from endemic areas.
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Esquistosomiasis mansoni , Esquistosomiasis , Quimiocina CXCL10/metabolismo , Quimiocina CXCL9/metabolismo , Quimiocinas/metabolismo , Citocinas/metabolismo , Humanos , Interleucina-33/metabolismo , Leucocitos Mononucleares , ARN Mensajero , Esquistosomiasis/metabolismo , Esquistosomiasis mansoni/epidemiología , Esquistosomiasis mansoni/metabolismoRESUMEN
Background: Social and environmental vulnerabilities contribute to the persistence and increase of Schistosomiasis, which has been a public health problem in Brazil and worldwide. In this study, we aimed to monitor the entry, installation, and maintenance of schistosomiasis transmission in an urban area on the Brazilian coast over two decades (2000-2010/2010-2020). Methods: This population-based cross-sectional study was conducted in Porto de Galinhas, state of Pernambuco, Brazil, to investigate the dynamics of schistosomiasis transmission in the urban area. Through 3 malacological and parasitological surveys and using geoprocessing technologies, schistosomiasis transmission foci (STF), as well as cases of the disease were identified and quantified. Statistical and geoprocessing tools were used to analyse the data. Findings: Overall, the number of STF decreased from 15 (2000) to 11 (2010) and then to 9 (2020). Although the infection ratio of snails in 2000 has decreased from 16·1% to 5·8% in 2010, we observed an increase to 7·2% in 2020. Additionally, 6,499 individuals were analysed (2012 in 2000; 2459 in 2010, and 2028 in 2020) and the prevalence of human infection has decreased over years, from 32·5% (2000), 16·6% (2010) to 8·8% (2020). The disorderly urbanization process was directly related to the spatial distribution of STF and schistosomiasis cases, causing a new scenario where people became infected by walking on unpaved and flooded streets. Interpretation: Although we observed a decreasing in schistosomiasis cases and STF, this NTD became a health problem related to urbanization in the study area. The challenge to overcome this new sort of transmission will require a greater understanding of the disorderly migration, spatial occupation, and degradation of the environment. Funding: National Council for Scientific and Technological Development (CNPq), Ministry of Science, Technology, Innovations and Communications, Brazil.
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OBJECTIVES: We measured the production of cytokines, chemokines and antibodies involved in allergic responses and sCD23 levels during Schistosoma mansoni infection. METHODS: Individuals (n = 164) were selected using the ISAAC questionnaire and parasitological exams. The subjects were divided as follows: those infected individuals with allergy-related symptoms (A-I), those with allergy-related symptoms only (A-NI); those only infected (NA-I); and those non-infected individuals without allergy-related symptoms (NA-NI). We used supernatants from cell culture (mitogenic stimulation) to measure cytokine and chemokine levels using cytometric bead arrays. Serum levels of anti-Ascaris lumbricoides (Asc) and anti-Blomia tropicalis IgE were measured using ImmunoCAP, and sCD23 was measured using ELISA. RESULTS: Schistosoma mansoni infection was associated with a lower risk of allergy-related symptoms. In A-I, there were higher levels of TNF-α, IL-10, IL-6, IFN-γ and CXCL8 than in NA-NI group, with TNF-α and IL-6 also at higher levels compared to A-NI group. Levels of IL-6, CXCL8, total and anti-Asc IgE, as well as the numbers of eosinophils, were higher in NA-I than in NA-NI, and the antibodies were also lower in A-NI than in NA-I group. In AI and NA-I, there was less production of CCL2 than in NA-NI. There were no differences in the levels of IL-2, IL-4, IL-17, CCL5, sCD23 and anti-Blomia IgE. CONCLUSIONS: Patients with allergy-related symptoms and infected (simultaneously) had higher levels of IL-10; due to the infection, there was increased production of IL-6 and CXCL8 and less CCL2. These data may characterize deviation to Th1 or attenuation of the Th2 response in allergy sufferers in areas endemic for schistosomiasis.
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Anticuerpos/inmunología , Quimiocinas/inmunología , Citocinas/inmunología , Hipersensibilidad Respiratoria/parasitología , Esquistosomiasis mansoni/inmunología , Adolescente , Adulto , Animales , Anticuerpos/sangre , Anticuerpos Antihelmínticos/sangre , Anticuerpos Antihelmínticos/inmunología , Brasil/epidemiología , Quimiocina CCL2/inmunología , Quimiocinas/sangre , Niño , Preescolar , Citocinas/sangre , Femenino , Humanos , Inmunoglobulina E , Masculino , Persona de Mediana Edad , Adulto JovenRESUMEN
Multiple epicenters of the SARS-CoV-2 pandemic have emerged since the first pneumonia cases in Wuhan, China, such as Italy, USA, and Brazil. Brazil is the third-most affected country worldwide, but genomic sequences of SARS-CoV-2 strains are mostly restricted to states from the Southeast region. Pernambuco state, located in the Northeast region, is the sixth most affected Brazilian state, but very few genomic sequences from the strains circulating in this region are available. We sequenced 101 strains of SARS-CoV-2 from patients presenting Covid-19 symptoms that reside in Pernambuco. Phylogenetic reconstructions revealed that all genomes belong to the B lineage and most of the samples (88%) were classified as lineage B.1.1. We detected multiple viral introductions from abroad (likely from Europe) as well as six local B.1.1 clades composed by Pernambuco only strains. Local clades comprise sequences from the capital city (Recife) and other country-side cities, corroborating the community spread between different municipalities of the state. These findings demonstrate that different from Southeastern Brazilian states where the epidemics were majorly driven by one dominant lineage (B.1.1.28 or B.1.1.33), the early epidemic phase at the Pernambuco state was driven by multiple B.1.1 lineages seeded through both national and international traveling.
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COVID-19/epidemiología , COVID-19/transmisión , Genoma Viral , Filogenia , SARS-CoV-2/genética , Brasil/epidemiología , Ciudades/epidemiología , Evolución Molecular , Genómica , Humanos , Estudios Longitudinales , Mutación , Nasofaringe/virología , Orofaringe/virología , SARS-CoV-2/aislamiento & purificaciónRESUMEN
In experimental autoimmune hepatitis (EAH) of Th1 profile, an extract of adult Ascaris suum worms (ASC) was previously found to deviate the immune response to a Th2/IL-10 pattern. Here, the effects of treatment with ASC on production of TGF-ß and the anti-Ascaris isotypes IgG1 and IgG2a in EAH were evaluated. EAH was induced in BALB/c mice, intravenously with concanavalin A. Two hours later, these animals received ASC (EAH+ASC group) or PBS vehicle (EAH group). IgG1 and IgG2a were evaluated 8 h, 24 h and 7 d after induction. TGF-ß was measured in a splenocyte culture at this last time. The isotype levels in the EAH group were low throughout the kinetics. In the EAH+ASC group, there was significant production of IgG1 at 24 h and 7 d, but of IgG2a only at 7 d. There was statistically greater production of TGF-ß in the EAH+ASC group. The higher levels of IgG1 and TGF-ß in this group suggest that an additional Th1 response control route exists in EAH, which needs to be investigated.
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Ascaris suum/inmunología , Hepatitis Autoinmune/parasitología , Inmunoglobulina G/biosíntesis , Factor de Crecimiento Transformador beta/biosíntesis , Animales , Anticuerpos Antihelmínticos/inmunología , Antígenos Helmínticos/inmunología , Modelos Animales de Enfermedad , Hepatitis Autoinmune/inmunología , Masculino , Ratones , Ratones Endogámicos BALB CRESUMEN
BACKGROUND: Maternal exposure to antibodies, cytokines or parasitic antigens during gestation may alter the degree of immune competence of offspring. Here we describe the production of cytokines and chemokines, and the ability to activation of the immune response in infants from mothers sensitized to helminths. METHODS: It were selected five infants born to helminth-seropositive mothers but who were negative for current helminth infection. Whole blood was cultured without stimulus, with phytohemagglutinin mitogen (5 µg/ml, 24 h) or with purified protein derivative (PPD) (1 µg/ml, 24 h), and the supernatant was assessed for presence of Th1/Th2 cytokines (IFN-γ, TNF-α, IL-10, IL-5, IL-4 and IL-2) and chemokines (CXCL10, CCL2, CXCL9, CCL5 and CXCL8) by cytometric bead array. RESULTS: All infants produced CCL5. Two infants demonstrated a mixed profile of Th1 (CXCL9) and Th2 (CCL2) chemokines in the presence of CXCL10, while one infant showed skewing towards Th2 without CXCL10 and two of them had been impaired immune response (children from sensitized to Schistosoma mansoni mothers). CONCLUSION: Infants with Th1 and Th2 profile chemokines demonstrated a good response to vaccination, indicated by CXCL10 levels, but not infants predominantly Th2-skewed profile. These results highlight that children from mothers sensitized to S. mansoni may lead to ineffective immune response to PPD, while mothers sensitized to Ascaris lumbricoides showed no such impairment.
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Antígenos Helmínticos/inmunología , Quimiocinas/inmunología , Citocinas/sangre , Inmunidad , Animales , Estudios de Casos y Controles , Citocinas/inmunología , Femenino , Helmintos/inmunología , Humanos , Lactante , Madres/estadística & datos numéricos , Embarazo , Complicaciones Infecciosas del Embarazo/inmunología , Complicaciones Infecciosas del Embarazo/parasitología , Células TH1/inmunología , Células Th2/inmunología , Adulto JovenRESUMEN
The relationship between the cellular immune response during Trichuris trichiura infection and asthma has not yet been established. In this study, the cytokines interleukin (IL)-2, IL-4, IL-6, IL-10, TNF-α, IFN-γ and IL-17A were evaluated in asthmatic children harboring T.â¯trichiura. For this assessment, asthmatic and non-asthmatic children (ISAAC questionnaire) were submitted to parasitological tests and blood samples were cultured (mitogen stimulation) for cytokine measurements in the supernatant. Asthma frequencies were similar in infected and uninfected children, but IL-4, IL-6, TNF-α and IL-10 levels were high in the infected asthmatic children. Additionally, infected non-asthmatic children exhibited high levels of these cytokines in relation to uninfected non-asthmatic children; however, cytokine levels were lower when compared with infected and asthmatic children. Therefore, T.â¯trichiura infection positively modulated the pro- and anti-inflammatory cytokines in asthmatic children, but a background of asthma seemed to narrow the production of cytokines induced by this helminth.
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Asma/parasitología , Citocinas/sangre , Tricuriasis/inmunología , Animales , Asma/inmunología , Brasil , Niño , Preescolar , Citocinas/inmunología , Femenino , Humanos , Hipersensibilidad/inmunología , Hipersensibilidad/parasitología , Inmunidad Celular , Masculino , TrichurisRESUMEN
In experimental autoimmune hepatitis (EAH) of Th1 profile, an extract of adult Ascaris suum worms (ASC) was previously found to deviate the immune response to a Th2/IL-10 pattern. Here, the effects of treatment with ASC on production of TGF-β and the anti-Ascaris isotypes IgG1 and IgG2a in EAH were evaluated. EAH was induced in BALB/c mice, intravenously with concanavalin A. Two hours later, these animals received ASC (EAH+ASC group) or PBS vehicle (EAH group). IgG1 and IgG2a were evaluated 8 h, 24 h and 7 d after induction. TGF-β was measured in a splenocyte culture at this last time. The isotype levels in the EAH group were low throughout the kinetics. In the EAH+ASC group, there was significant production of IgG1 at 24 h and 7 d, but of IgG2a only at 7 d. There was statistically greater production of TGF-β in the EAH+ASC group. The higher levels of IgG1 and TGF-β in this group suggest that an additional Th1 response control route exists in EAH, which needs to be investigated.(AU)
Na hepatite autoimune experimental (HAE) de perfil Th1, o extrato de vermes adultos Ascaris suum (ASC) desviou a resposta imune para um padrão Th2/IL-10. Neste trabalho, foram avaliados os efeitos do tratamento com ASC na produção TGF-β e dos isótipos de IgG1 e IgG2a anti-Ascaris na HAE. Esta foi induzida em camundongos BALB/c intravenosamente com Concanavalina A. Após duas horas, os animais receberam ASC (grupo HAE+ASC) ou veículo PBS (grupo HAE). IgG1 e IgG2a foram avaliados em 8 horas, 24 horas e 7 dias após indução. TGF-β foi mensurado em cultura de esplenócitos nesse último tempo. Os níveis dos isótipos no grupo HAE foram baixos durante toda a cinética. No grupo HAE+ASC, houve produção significativa de IgG1 em 24 horas e 7 dias, mas somente em 7 dias para IgG2a. A produção de TGF-β foi estatisticamente maior no grupo HAE+ASC. Níveis mais altos de IgG1 e TGF-β nesse grupo sugerem uma via adicional de controle da resposta Th1 na HAE que precisa ser investigada.(AU)
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Animales , Inmunomodulación , Ascaris suum/inmunología , Hepatitis Autoinmune , Factor de Crecimiento Transformador betaRESUMEN
Abstract In experimental autoimmune hepatitis (EAH) of Th1 profile, an extract of adult Ascaris suum worms (ASC) was previously found to deviate the immune response to a Th2/IL-10 pattern. Here, the effects of treatment with ASC on production of TGF-β and the anti-Ascaris isotypes IgG1 and IgG2a in EAH were evaluated. EAH was induced in BALB/c mice, intravenously with concanavalin A. Two hours later, these animals received ASC (EAH+ASC group) or PBS vehicle (EAH group). IgG1 and IgG2a were evaluated 8 h, 24 h and 7 d after induction. TGF-β was measured in a splenocyte culture at this last time. The isotype levels in the EAH group were low throughout the kinetics. In the EAH+ASC group, there was significant production of IgG1 at 24 h and 7 d, but of IgG2a only at 7 d. There was statistically greater production of TGF-β in the EAH+ASC group. The higher levels of IgG1 and TGF-β in this group suggest that an additional Th1 response control route exists in EAH, which needs to be investigated.
Resumo Na hepatite autoimune experimental (HAE) de perfil Th1, o extrato de vermes adultos Ascaris suum (ASC) desviou a resposta imune para um padrão Th2/IL-10. Neste trabalho, foram avaliados os efeitos do tratamento com ASC na produção TGF-β e dos isótipos de IgG1 e IgG2a anti-Ascaris na HAE. Esta foi induzida em camundongos BALB/c intravenosamente com Concanavalina A. Após duas horas, os animais receberam ASC (grupo HAE+ASC) ou veículo PBS (grupo HAE). IgG1 e IgG2a foram avaliados em 8 horas, 24 horas e 7 dias após indução. TGF-β foi mensurado em cultura de esplenócitos nesse último tempo. Os níveis dos isótipos no grupo HAE foram baixos durante toda a cinética. No grupo HAE+ASC, houve produção significativa de IgG1 em 24 horas e 7 dias, mas somente em 7 dias para IgG2a. A produção de TGF-β foi estatisticamente maior no grupo HAE+ASC. Níveis mais altos de IgG1 e TGF-β nesse grupo sugerem uma via adicional de controle da resposta Th1 na HAE que precisa ser investigada.
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Animales , Masculino , Conejos , Inmunoglobulina G/biosíntesis , Factor de Crecimiento Transformador beta/biosíntesis , Ascaris suum/inmunología , Hepatitis Autoinmune/parasitología , Anticuerpos Antihelmínticos/inmunología , Hepatitis Autoinmune/inmunología , Modelos Animales de Enfermedad , Ratones Endogámicos BALB C , Antígenos Helmínticos/inmunologíaRESUMEN
Laboratory maintenance of the Schistosoma mansoni cycle is necessary for developing studies regarding the diagnosis, treatment and control of schistosomiasis. Within this perspective, it is paramount that mice infected by the parasite should present a minimum survival of six months. However, between October 2016 and May 2017, early deaths were observed among infected animals kept in the vivarium of the Schistosomiasis Reference Service of IAMFIOCRUZ. Therefore, the purpose of the present study was to present the results obtained after investigating the main cause of death among these animals. To achieve this, animals that died or that needed to be euthanized due to clinical distress caused by parasite infection were necropsied to investigate the cause of death and clinical condition. Fragments from the intestines, mesenteric vessels and livers were removed and were subjected to histopathological studies. In addition, mouse feces were collected and analyzed using the hydrogen peroxide reaction to detect occult blood. Over an eight-month period, 70 deaths were noted. Forty two animals presented mesenteric ischemia, a vascular insufficiency syndrome that causes a reduction in the nutrient supply to the intestinal viscera. There is, therefore, a need to reduce the infective parasite load in mice to increase their survival, reduce distress caused by the infection and ensure maintenance of the S. mansoni cycle, thus enabling continuity of scientific studies on this parasitosis.
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Animales , Ratones , Schistosoma mansoni , Isquemia Mesentérica , RatonesRESUMEN
ABSTRACT Introduction: Kato-Katz is a laboratory method recommended by the Brazilian Ministry of Health (BMH) and the World Health Organization (WHO) as the gold standard for the diagnosis of human infection by Schistosoma mansoni. The method has great clinical and epidemiological relevance because it allows the parasite load quantification of the infected patient by calculating the number of eggs per gram (EPG) of feces. This classification may also be used to estimate the intensity of infection in the communities, to measure the impact of disease control measures, as well as to establish quality parameters for reading the slides. Objective: To describe the correct laboratory procedures for the parasitological diagnosis of S. mansoni infection by the Kato-Katz method based on the quality control protocol established by the Laboratory and Reference Service in Schistosomiasis/Centro de Pesquisa Aggeu Magalhães (CPqAM)/Fundação Oswaldo Cruz (Fiocruz)/BMH. Methods: We describe: 1) the technical steps for fecal sample preparation and reading the slides; 2) the technical limitations; 3) the standard operating procedure (SOP) to be adopted by laboratories; 4) the methodology for the internal and external quality control of the reading slides results; and 5) the tolerance limits accepted for such control. Conclusion: This study provides the laboratory which performs the diagnosis of schistosomiasis using the Kato-Katz method with parameters to implement a diagnostic service that can be evaluated internally and externally. The establishment of a quality protocol enables the comparison of data and the identification of failures in the operational procedure, which can be corrected by training personnel and taking actions for the problems identified.
RESUMO Introdução: O Kato-Katz é o método laboratorial adotado pelo Ministério da Saúde (MS) e pela Organização Mundial da Saúde (OMS) como padrão-ouro para o diagnóstico da infecção humana pelo Schistosoma mansoni, sendo uma ferramenta de relevância clínica e epidemiológica, visto que permite classificar a carga parasitária do indivíduo infectado pelo cálculo de ovos por grama de fezes (OPG). Essa classificação pode também ser utilizada para estimar a intensidade da infecção nas comunidades, mensurar o impacto de medidas de controle da doença bem como estabelecer parâmetros de qualidade para a leitura das lâminas. Objetivo: Descrever os procedimentos laboratoriais corretos para o diagnóstico parasitológico da infecção pelo S. mansoni pelo método Kato-Katz a partir do protocolo de controle de qualidade estabelecido pelo Laboratório e pelo Serviço de Referência em Esquistossomose/Centro de Pesquisa Aggeu Magalhães (CPqAM)/Fundação Oswaldo Cruz (Fiocruz)/MS. Método: São descritas: 1) as etapas técnicas para o preparo das amostras de fezes e a leitura das lâminas; 2) as limitações da técnica; 3) o procedimento operacional padrão (POP) a ser adotado pelos laboratórios; 4) a metodologia para o controle de qualidade interno e externo da leitura das lâminas; e 5) os limites de tolerância aceitos para tal controle. Conclusão: Este trabalho instrumentaliza os laboratórios que realizam o diagnóstico da esquistossomose pelo método Kato-Katz com parâmetros para implantar um serviço diagnóstico passível de ser avaliado interna e externamente. O estabelecimento de um protocolo de qualidade viabiliza a comparação de dados e a identificação de falhas no procedimento operacional, que poderão ser corrigidas por meio de capacitação de pessoal e tomada de medidas para os problemas identificados.
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RESUMO: Introdução: A esquistossomose é considerada uma endemia em Vitória de Santo Antão, Pernambuco, o qual apresenta há décadas altas incidências e prevalências para essa doença. Nesse município ocorre a transmissão clássica da doença por meio do contato da população de áreas rurais com águas contaminadas durante o desenvolvimento de suas atividades de vida diárias. Recentemente surgiram indícios da presença do caramujo vetor na área urbana da cidade, o que pode configurar um novo modelo de transmissão para esquistossomose nesse município. Objetivo: Identificar novo cenário epidemiológico para ocorrência da transmissão urbana da esquistossomose em Vitória de Santo Antão, Pernambuco. Métodos: Foi conduzido um inquérito malacológico, investigando-se todas as coleções hídricas do perímetro urbano quanto à presença do caramujo vetor da esquistossomose (Biomphalaria spp.). Os caramujos coletados foram examinados para identificação taxonômica e de infecção pelo Schistosoma mansoni. Todos os criadouros (CRs) foram georreferenciados para construção de mapas de risco por meio dos software TrackMaker-Pro e ArcGIS. Resultados: Foram identificados 22 CRs da espécie Biomphalaria straminea, nos quais foram coletados 1.704 caramujos. Desses CRs, um foi identificado como foco de transmissão da doença e sete como focos potenciais para transmissão. Os mapas construídos identificaram duas áreas de risco para transmissão urbana da esquistossomose, bem como áreas de expansão dos CRs, configurando um aumento no risco de transmissão para a população. Conclusão: Os resultados constatam a existência de um novo cenário epidemiológico, no qual a possibilidade de transmissão urbana dessa doença foi confirmada.
ABSTRACT: Introduction: Schistosomiasis is considered an endemic disease in Vitória de Santo Antão, Pernambuco, a district which has presented both high incidence and prevalence of it for decades. Poor environmental conditions lead to contamination of water sources in rural areas, which are used by the population during daily activities, resulting in typical transmission. Recently, there has been evidence of vector snails in urban areas, which could set a new model for schistosomiasis transmission in this district. Objective: To identify the new epidemiological situation for the urban transmission of schistosomiasis in Vitória de Santo Antão, Pernambuco. Methods: A malacological survey was conducted in all water sources in the city limits to investigate schistosomiasis vector snails (Biomphalaria spp.). The collected snails were examined for taxonomic identification and Schistosoma mansoni infection. All breeding sites were georeferenced to build risk maps through the TrackMaker PRO program and ArcGIS software. Results: We identified 22 Biomphalaria straminea breeding sites and collected 1,704 snails. One of these breeding sites was identified as a source of transmission and seven as potential sources of transmission. The designed maps identified two risk areas of urban transmission of schistosomiasis and expansion areas for breeding sites, establishing an increased risk of transmission to the population. Conclusion: This study verified the existence of a new epidemiological situation in which the possibility of the urban transmission of the disease was confirmed.
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Humanos , Animales , Biomphalaria/parasitología , Vectores de Enfermedades , Esquistosomiasis mansoni , Esquistosomiasis/transmisión , Salud Urbana , Brasil , BosquesRESUMEN
Schistosoma mansoni antigens in the early life alter homologous and heterologous immunity during postnatal infections. We evaluate the immunity to parasite antigens and ovalbumin (OA) in adult mice born/suckled by schistosomotic mothers. Newborns were divided into: born (BIM), suckled (SIM) or born/suckled (BSIM) in schistosomotic mothers, and animals from noninfected mothers (control). When adults, the mice were infected and compared the hepatic granuloma size and cellularity. Some animals were OA + adjuvant immunised. We evaluated hypersensitivity reactions (HR), antibodies levels (IgG1/IgG2a) anti-soluble egg antigen and anti-soluble worm antigen preparation, and anti-OA, cytokine production, and CD4+FoxP3+T-cells by splenocytes. Compared to control group, BIM mice showed a greater quantity of granulomas and collagen deposition, whereas SIM and BSIM presented smaller granulomas. BSIM group exhibited the lowest levels of anti-parasite antibodies. For anti-OA immunity, immediate HR was suppressed in all groups, with greater intensity in SIM mice accompanied of the remarkable level of basal CD4+FoxP3+T-cells. BIM and SIM groups produced less interleukin (IL)-4 and interferon (IFN)-g. In BSIM, there was higher production of IL-10 and IFN-g, but lower levels of IL-4 and CD4+FoxP3+T-cells. Thus, pregnancy in schistosomotic mothers intensified hepatic fibrosis, whereas breastfeeding diminished granulomas in descendants. Separately, pregnancy and breastfeeding could suppress heterologous immunity; however, when combined, the responses could be partially restored in infected descendants.
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Animales Lactantes/inmunología , Anticuerpos Antihelmínticos/inmunología , Granuloma de Cuerpo Extraño/inmunología , Inmunidad Humoral/fisiología , Parasitosis Hepáticas/inmunología , Esquistosomiasis mansoni/inmunología , Adyuvantes Inmunológicos , Animales , Animales Recién Nacidos , Animales Lactantes/parasitología , Linfocitos T CD4-Positivos/parasitología , Cercarias/inmunología , Ensayo de Inmunoadsorción Enzimática , Femenino , Citometría de Flujo , Factores de Transcripción Forkhead/sangre , Granuloma de Cuerpo Extraño/parasitología , Granuloma de Cuerpo Extraño/patología , Inmunidad Heteróloga/fisiología , Inmunoglobulina G/sangre , Interferón gamma/sangre , Interleucina-10/sangre , Interleucina-4/sangre , Cirrosis Hepática/inmunología , Cirrosis Hepática/parasitología , Parasitosis Hepáticas/patología , Masculino , Ratones , Madres , Ovalbúmina/inmunología , Embarazo , Schistosoma mansoni/inmunología , Bazo/inmunología , Bazo/patologíaRESUMEN
Schistosoma mansoni antigens in the early life alter homologous and heterologous immunity during postnatal infections. We evaluate the immunity to parasite antigens and ovalbumin (OA) in adult mice born/suckled by schistosomotic mothers. Newborns were divided into: born (BIM), suckled (SIM) or born/suckled (BSIM) in schistosomotic mothers, and animals from noninfected mothers (control). When adults, the mice were infected and compared the hepatic granuloma size and cellularity. Some animals were OA + adjuvant immunised. We evaluated hypersensitivity reactions (HR), antibodies levels (IgG1/IgG2a) anti-soluble egg antigen and anti-soluble worm antigen preparation, and anti-OA, cytokine production, and CD4+FoxP3+T-cells by splenocytes. Compared to control group, BIM mice showed a greater quantity of granulomas and collagen deposition, whereas SIM and BSIM presented smaller granulomas. BSIM group exhibited the lowest levels of anti-parasite antibodies. For anti-OA immunity, immediate HR was suppressed in all groups, with greater intensity in SIM mice accompanied of the remarkable level of basal CD4+FoxP3+T-cells. BIM and SIM groups produced less interleukin (IL)-4 and interferon (IFN)-g. In BSIM, there was higher production of IL-10 and IFN-g, but lower levels of IL-4 and CD4+FoxP3+T-cells. Thus, pregnancy in schistosomotic mothers intensified hepatic fibrosis, whereas breastfeeding diminished granulomas in descendants. Separately, pregnancy and breastfeeding could suppress heterologous immunity; however, when combined, the responses could be partially restored in infected descendants.
Asunto(s)
Animales , Femenino , Masculino , Ratones , Embarazo , Animales Lactantes/inmunología , Anticuerpos Antihelmínticos/inmunología , Granuloma de Cuerpo Extraño/inmunología , Inmunidad Humoral/fisiología , Parasitosis Hepáticas/inmunología , Esquistosomiasis mansoni/inmunología , Adyuvantes Inmunológicos , Animales Recién Nacidos , Animales Lactantes/parasitología , /parasitología , Cercarias/inmunología , Ensayo de Inmunoadsorción Enzimática , Citometría de Flujo , Factores de Transcripción Forkhead/sangre , Granuloma de Cuerpo Extraño/parasitología , Granuloma de Cuerpo Extraño/patología , Inmunidad Heteróloga/fisiología , Inmunoglobulina G/sangre , Interferón gamma/sangre , /sangre , /sangre , Cirrosis Hepática/inmunología , Cirrosis Hepática/parasitología , Parasitosis Hepáticas/patología , Madres , Ovalbúmina/inmunología , Schistosoma mansoni/inmunología , Bazo/inmunología , Bazo/patologíaRESUMEN
INTRODUCTION:: Schistosomiasis is considered an endemic disease in Vitória de Santo Antão, Pernambuco, a district which has presented both high incidence and prevalence of it for decades. Poor environmental conditions lead to contamination of water sources in rural areas, which are used by the population during daily activities, resulting in typical transmission. Recently, there has been evidence of vector snails in urban areas, which could set a new model for schistosomiasis transmission in this district. OBJECTIVE:: To identify the new epidemiological situation for the urban transmission of schistosomiasis in Vitória de Santo Antão, Pernambuco. METHODS:: A malacological survey was conducted in all water sources in the city limits to investigate schistosomiasis vector snails (Biomphalaria spp.). The collected snails were examined for taxonomic identification and Schistosoma mansoni infection. All breeding sites were georeferenced to build risk maps through the TrackMaker PRO program and ArcGIS software. RESULTS:: We identified 22 Biomphalaria straminea breeding sites and collected 1,704 snails. One of these breeding sites was identified as a source of transmission and seven as potential sources of transmission. The designed maps identified two risk areas of urban transmission of schistosomiasis and expansion areas for breeding sites, establishing an increased risk of transmission to the population. CONCLUSION:: This study verified the existence of a new epidemiological situation in which the possibility of the urban transmission of the disease was confirmed.
Asunto(s)
Biomphalaria/parasitología , Vectores de Enfermedades , Esquistosomiasis mansoni , Esquistosomiasis/transmisión , Salud Urbana , Animales , Brasil , Bosques , HumanosRESUMEN
Adult worm extract from Ascaris suum (Asc) has immunosuppressive activity and elicits Th2/IL-4/IL-10 response. This study evaluated the prophylactic and therapeutic effect of Asc in a murine model of concanavalin A (ConA)-induced autoimmune hepatitis (AIH). BALB/c mice received ConA, iv, (20 mg/kg), and three groups of animals were formed: (1) AIH, received only ConA; (2) AIH + Asc prophylactic, treated with Asc (1 mg/ml), ip, 30 min before of the AIH; and (3) AIH + Asc therapeutic, treated with Asc 2 h after the AIH. Plasma transaminase and immunoglobulins (measured at 8 and 24 h and 7 days after treatment) and cytokine production (IL-4, IL-10, IL-13, and IFN-γ) by splenocytes upon ConA and Asc stimulus were compared. The livers were weighed and examined histologically. In the AIH group, there was an increase in liver weight, transaminase levels, and total immunoglobulins. These parameters were reduced by 8-24 h and 7 days in the prophylactic group, but in the therapeutic group, only on day 7. The survival rate of mice in the AIH group was 38.5%, compared to 67% in the therapeutic Asc group. The survival rate of the animals with AIH that were prophylactically treated with Asc was 100%. A decrease of cellular infiltration and high levels of IL-4, IL-10, and IL-13 were induced by Asc. An increase of liver fibrosis was also observed, but with less intensity with prophylactic treatment. Thus, the Ascaris components have an inhibitory effect on AIH, with an intense Th2 immune response.