Concanavalin A with the property enhances T-cell activation have been known. BALB/c mice infected with Angiostrongylus cantonensis administrated with different doses 2.5mg/kg, 5mg/kg, 10mg/kg and vehicle from 7th day to 10th day p.i. and administrated with a single dose 5mg/kg at different period started from 7th day to 10th day, 14th day to 17th day,21th day to 24th day p.i. Effects of concanavalin A on Angiostrongylus cantonensis infected mice was determined by counting the percentage of eosinophil in blood, measuring the worm length, protein density in cerebro-spinal fluid, ELISA, and histological observation. Concanavalin A injection promotes eosinophila in Angiostrongylus cantonensis infected mice, antibody response and inhibits larvae growth. Protein density in CSF decrease when 10mg/kg dose given and period given from14th day to 17th day,period given from 21th day to 24th day. And hemorrhage in brain tissue was reduced.

The IgE response increased in hosts infected with Angiostrongylus cantonensis. This study analyzes IgE responses appeared in different body fluids of mice infected with A. cantonensis. BALB/c, CBA, C57BL/6 and C57BL/10 strain of mice were used in the experiment. BALB/c mice exhibited the highest susceptibility to A. cantonensis, CBA mice showed intermediate susceptibility, while C57BL/6 and C57BL/10 mice showed the lowest susceptibility. Analyses of specific antibodies against L5 antigens, and ES antigens in mice showed antibodies appeared in sera and in cerebrospinal fluids(CSF), especially IgE antibodies, were related to the worm rejection happened in mouse brain. However, antibodies appeared in ascites had no influence. The level of IgE increased in brain of susceptible BALB/c mice is much lower than that increased in other strain. Our data suggested that IgE response elicited in brain might be much more important than that in serum. Western blot analyses showed that among antigens extracted from L5 both IgE and IgG responses were positive in CSF. IgE antibodies against a 105 kDa protein of L5 antigen in C57BL/10 mice appeared in third week after infection, while in BALB/c mice appeared in fifth week. IgG antibodies against 105 kDa protein appeared in the identical weeks. Our results indicated that 105 kDa protein was related to specific antibodies responses about protective function of mice infected with A. cantonensis. Further researches will be required to understand function and characterization of this protein.

Reproduction de : Thèse d'exercice : Médecine. Médecine générale : Nantes : 2008. / Bibliogr.

Düsseldorf, Universiẗat, Diss., 2004.

Made available in DSpace on 2016-07-13T19:05:35Z (GMT). No. of bitstreams: 2 danielle_vasconcelos_ioc_dout_2016.pdf: 14478401 bytes, checksum: 974155eb21c57b43bb5e39f47c88c29a (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2016 / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil / Introdução: O Angiostrongylus costaricensis é um nematódeo que causa a angiostrongilíase abdominal. O ciclo de vida deste parasita envolve um hospedeiro invertebrado (intermediário) e um hospedeiro vertebrado mamífero (definitivo). Na natureza, um dos principais hospedeiros definitivos deste parasita é o roedor Sigmodon hispidus. O nematódeo instala-se na artéria mesentérica superior do hospedeiro definitivo, causando inicialmente uma periarterite eosinofílica e posteriormente um trombo. Histopatologicamente, uma tríade de achados fundamentais define esta patologia: massivo infiltrado eosinofílico; vasculite eosinofílica e reação granulomatosa. Apesar da importância clínica, pouco se sabe acerca dessa patologia. Objetivos: Caracterizar as lesões histopatológicas do território vascular e intestinal; acompanhar a dinâmica hematológica periférica e central; avaliar colesterolemia total e a presença de bactérias na lesão de S. hispidus infectados com A. costaricensis. Material e Métodos: S. hispidus infectados com A. costaricensis foram eutanasiados em 30, 50, 90 e 114 dpi e foram coletados o intestino e mesentério (incluindo a artéria cecal). Os tecidos foram fixados em formalina Millonig de Carson e, histologicamente processados para microscopia de luz ou imunohistoquímica. O sangue também foi coletado através de punção cardíaca ou do plexo braquial e utilizado para realização de hemograma e análise da colesterolemia total e análise microbiológica. A lesão local também foi coletada para análise microbiológica Resultados e discussão: Em tempos precoces (30 e 50 dpi), uma grande quantidade de larvas e ovos viáveis foram encontrados na vasa vasorum da artéria cecal levando a um intenso infiltrado eosinofílico na adventícia do vaso, formando granulomas. Os parasitas associados ao infiltrado inflamatório promoveram uma vasculite eosinofílica centrípeta com destruição das lâminas elásticas. A íntima perdeu as características celulares básicas, tornando-se desorganizada ou até mesmo desaparecendo. Com a evolução temporal da doença, há um aumento da severidade em tempos tardios (90 e 114 dpi), onde um processo aterosclerótico instala-se. O trombo é composto majoritariamente por fibrina, além de ferro e uma substância xantomatosa, positiva para Vermelho do Congo. Neste momento, o infiltrado eosinofílico cessa e há uma predominância de macrófagos carreadores dessa substância xantomatosa e ferro. As lâminas elásticas de artérias com vermes estão destruídas, havendo uma grande desestruturação do vaso. No sangue, em 90dpi há um aumento do número de leucócitos totais, diminuição do número e aumento do tamanho das plaquetas. Nesse ponto, a colesterolemia total dos animais não tem modificação em relação aos controles. Nesses tempos tardios, observa-se um nítido aumento da atividade da medula, com predominante eosinofilia e aumento do número de megacariócitos. É possível que o desenvolvimento dessas lesões esteja relacionado à presença de bactérias grampositivas presentes no A. costaricesis. Conclusões: Nossos dados sugerem que o modelo de infecção do S. hispidus pelo A. costaricensis, pode ser útil no estudo de vasculites autoimunes, bem como aterosclerose / Abstract: Introduction: Angiostrongylus costaricensis is a roundworm which causes the abdominal angiostrongyliasis. This parasite\2019s life cycle involves an invertebrate host (intermediary) and a vertebrate host (definitive). In nature, the usual definitive host for this parasite is the rodent Sigmodon hispidus. Adult worms install in the upper mesenteric artery of the definitive host, causing in the beginning an eosinophilic polyarteritis and then a thrombus. Histopathologically, a triad of essential finding defines this pathology as: massive eosinophilic infiltrate; eosinophilic vasculitis and the granulomatous reaction. Despite the clinical importance, we know just a few about this pathology. Objectives: To characterize the histopathological lesions of the vascular and intestinal territories; to follow the peripheral and central hematologic dynamics; to evaluate the total cholesterol and the bacteria presence in the lesion on the infected S. hispidus by A. costaricensis. Material and Methods: S. hispidus infected by A. costaricensis were euthanized in 30, 50, 90 and 114 dpi and were colected the intestine and mesentery (including the cecal artery). Tissues were fixed in Carson\2019s Millonig formalin and, histopathologically processed to light microscopy or immunohistochemistry. The blood was also gathered through the cardiac puncture or from the brachial plexus and used to the CBC conduction and analysis of total cholesterol and microbiological analysis. The local lesion was also gathered to microbiological analysis Results and discussion: In early times (30 and 50 dpi), a big quantity of larvae and viable eggs were found at the vasa vasorum of the cecal artery leading to an intense eosinophilic infiltrate at the adventitious vessel, forming granulomas. The parasites associated to the inflammatory infiltrate promote an eosinophilic vasculitis centripetal with the elastic blades destruction. The inner loses the basic cell characteristics, becoming disorganized or even vanishing. With the disease\2019s temporal evolution, there is an increase at the disease severity in later times (90 and 114 dpi), when an atherosclerotic process gets installed. A thrombus composed mainly by fibrin, iron and a xanthomatous substance, positive to Red Congo. At this moment, the eosinophilic infiltration ceases and there is a predominance of macrophage carriers of this xanthomatous substance and iron. The elastic arteries blades with worms are destroyed bringing a big vessel disruption. In the blood, in 90dpi there is an increasing on the total leukocytes number, decreasing on the platelets number and an increasing on their size. At this point, the animal total cholesterol has no change in relation to the controls. At those later moments, it is observed a clear increase on the marrow activity, with predominant eosinophilia and megakaryocytes number increasing. It is possible that those lesions development may be related to the gram positive bacteria present at A. costaricesis. Conclusions: Our data suggest that the model of the S. hispidus l infected by A. costaricensis can be usual by the atherosclerosis study

This study evaluates the efficiency of albendazole against different stages of Angiostrongylus cantonensis in Sprague-Dawley rats. Using a single oral dose of albendazole of 10 mg/kg at 0 or 1 or 2 or 3 weeks post-infection (PI) showed that the drug in 0 or 1 weeks PI was more effective in the larval infection. Compared to those in non-treated control, the rats treated 0 or 1 weeks PI showed significant reductions not only in the relative wet weight of heart and lungs (g/100 g body weight), but also in the mean number of adults recovered. With regard to single oral doses of 10 or 50 or 100 mg/kg at weekly intervals from 6 weeks PI, the rats treated 100 mg/kg in 2 weeks post-treatment (PT) showed significant reduction in the L1 in its feces and the amounts were lastingly few. In the rats treated 10 or 50 mg/kg, larval output reappeared from 4 weeks PT. The histomorphology of granuloma formation was also investigated in the lungs of SD rats infected with A. cantonensis¡@by paraffin section. It was observed that granulomatous fibros was strongly associated with the number of L1 in the lungs. IgG and IgE antibodies specific for antigens from L1, L5, male worms or female adults of A. cantonensis were respectively measured by an enzyme-linked immunosorbent assay (ELISA). The specific IgE response to antigens from L1 or female adults of A. cantonensis in the serum reached the maximum at 1 and 5 weeks PI. Similarly, the eosinophil response also revealed remarkable peaks at 1 and 5 weeks PI. These results might prove that the increases of IgE titer and eosinophil are related to antibody-dependent cell mediated cytotoxicity (ADCC).

Made available in DSpace on 2016-03-08T14:05:17Z (GMT). No. of bitstreams: 2 juberlan_garcia_ioc_dout_2014.pdf: 5272747 bytes, checksum: 0a5731978e1e998b6728ba6318bbdb14 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2016-02-23 / A meningite eosinofílica é uma doença caracterizada pelo aumento de eosinófilos no líquido cefalorraquidiano (LCR), cuja causa mais comum é a invasão do sistema nervoso central por helmintos. Entre os helmintos, destaca-se o Angiostrongylus cantonensis. O roedor Rattus norvegicus é o hospedeiro natural definitivo e o homem atua como hospedeiro acidental e que pode se infectar através da ingestão de moluscos crus ou mal cozidos ou alimentos contaminados com a larva L3. Recentemente, no Brasil houve quatro casos de meningite eosinofílica, devido à ingestão ou manipulação de Achatina fulica infectada. Foi realizado uma avaliação das alterações metabólicas, fisiológicas e histopatológicas em R. norvegicus experimentalmente infectados com 100 larvas L3 de A. cantonensis e o estudo do efeito da infecção concomitante por Echinostoma paraensei e A. cantonensis sobre a atividade das aminotransferases ALT e AST, na concentração de proteínas totais, ácido úrico, ureia e análise histopatológica para compreender o efeito da infecção concomitante em Biomphalaria glabrata Os resultados obtidos demonstraram que a infecção causada por A. cantonensis em R. norvegicus promoveu alterações metabólicas hepáticas e histopatológicas, caracterizadas pelo aumento da atividade das enzimas ALT, AST, GGT e ALKP, alteração no metabolismo glicídico, presença de infiltrado inflamatório, alterações hematológicas significativas, além disso promoveu alterações histopatológicas no parênquima pulmonar demonstraram nos roedores a ocorrência de áreas de necrose e extensa fibrose, podendo estar diretamente relacionada com o desenvolvimento de hipóxia celular. Os moluscos infectados apresentaram alterações nos níveis das aminotransferases e aumento na excreção de produtos nitrogenados, os resultados histopatológicos mostraram uma mudança na distribuição de A. cantonensis na presença de E. paraensei, indicando que a presença deste trematódeo pode interferir na dinâmica do A. cantonensis. A infecção por A. cantonensis causou alterações metabólicas e histopatológicas nos roedores / E osinophilic meningitis is a disease characterized by increased eosinophils in the cerebrospinal fluid (CSF), which is the most common ly cause d by invasion of the central nervous system by helminths , as occurs in Angiostrongylus cantonensis infections . The rodent Rattus norvegicus is the definitive natural host and humans act as accidental host s and can become infected by eating raw or undercooked snail s or food contaminated with infective L 3 larvae. Recently in Brazil there have been four cases of eosinophilic meningitis due to ingestion of infected Achatina fulica . An evaluation of metabolic, p hysiological and pathological changes in R. norvegicus experimentally infected with 100 L 3 larvae of A. cantonensis and study the effect of co - infection with Echinostoma paraensei and A. cantonensis on the activity of aminotransferases ALT and AST was perf ormed in the concentration total protein, uric acid, urea and histopathologic analysis to understand the effect of co - infection in Biomphalaria glabrata . The results showed that the infection caused by A. cantonensis in R. norvegicus promoted histopathological changes in liver metabolism is characterized by increased activity of ALT, AST, GGT and ALKP enzymes in glucose metabolism, inflammatory infiltrate, hematological changes significant in addition in addition histopathological alt erations in lung parenchyma in rodents have shown the occurrence of extensive areas of necrosis and fibrosis, which can be directly related to the development of cellular hypoxia. The infected snails showed altered levels of aminotransferases and increased excretion of nitrogen products, histopathological findings showed a change in the distribution of A. cantonensis in the presence of E. paraensei, indicating that the presence of this trematode may interfere with the dynamics of A. cantonensis .

Abstract In order to determine whether antigens prepared from adult intestine of Angiostrongylus cantonensis have potential to induce a protective immunity in the rodent hosts, somatic antigens extracted from male and female adults as well as gut antigens isolated from female adults were used to immunize rats against A. cantonensis infection. A 14% reduction in L5 recorvery from brain as well as a 15% reduction in adult recovery from pulmonary artery were achieved in the immunized rats when compared with the control group after infection with 50 larvae. The length of worms recovered from immunized rats was shorter than that in other groups. The larvae recovered from fecal materials in immunized rats were also reduced. In cell proliferation test, the stimulation index of gut antigens increased with times of immunization and exhibited the highest values. However, serum IgG titers were not correlated with protective immune responses. A 84 kDa protein contained in all antigen preparations was recognized by immune serum against gut antigens. Strong positive reactions were detected by indirect immunofluorescent assay in the internal musculature of the body, gut and reproductive tract wall and gut lumen. The composition of gut antigens was similar to that of gut membrane proteins. Immune sera recognized several major gut proteins were also appeared on gut membrane proteins. Further studies are required to provide evidence that gut membrane proteins play in the protective immune response against A. cantonensis infections, including the 84 kDa protein.

Made available in DSpace on 2016-02-26T13:38:42Z (GMT). No. of bitstreams: 2 taina_monte_ioc_mest_2014.pdf: 2518433 bytes, checksum: 7691ab89b672f7a5b9084cbb9979957c (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2016-02-23 / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil / Angiostrongylus cantonensis é responsável por causar meningoencefalite eosinofílica em humanos e casos já foram registrados em diversas partes do mundo incluindo o Brasil (ES, PE e SP). Nesse estudo, relatamos a variabilidade genética entre isolados de A. cantonensis do Brasil utilizando sequências do gene mitocondrial COI. Foram identificados três haplótipos brasileiros de A. cantonensis, baseados em oito haplótipos conhecidos (ac1-ac8). O haplótipo brasileiro ac5 ficou agrupado com isolados do Japão e o haplótipo brasileiro ac8 (isolados do RJ, SP, PA e PE) formaram um clado distinto. Foi relatado um novo haplótipo brasileiro, haplótipo ac9, o qual se encontra intimamente relacionado com os haplótipos da China (ac6) e do Japão (ac7). Dois isolados brasileiros de A. cantonensis, Olinda e Caju (haplótipos ac8 e ac9, respectivamente) relatados no presente estudo, tiveram sua biologia e morfologia caracterizadas após infecção experimental. Foi observada diferença significativa com maior carga parasitária recuperada nos isolados de Caju e um número significativamente maior de larvas L1 eliminadas nas fezes no início do período patente. Entretanto, quando comparado o total de larvas eliminadas não foi verificada diferença significativa entre os dois isolados O isolado de Caju apresentou diferença significativa na proporção entre espécimes fêmeas e machos (0,64:1), enquanto que o mesmo não foi observado para o isolado de Olinda (1,16:1). A análise morfométrica revelou que os espécimes machos e fêmeas do isolado de Olinda foram significativamente maiores com relação aos caracteres analisados quando comparados com os espécimes de Caju. A análise morfológica evidenciou pequena variação no nível das bifurcações que unem os raios laterais no lobo direito da bolsa copuladora, entre os dois isolados. A variação genética observada apoia a hipótese que o aparecimento do parasito no Brasil é um resultado de múltiplas introduções de ratos parasitados e pelo molusco Achatina fulica, o qual contribui para a dispersão. As variações biológicas, morfológicas e morfométricas entre os dois haplótipos estudados, reforçam a variação observada pelo marcador COI e pela possível influência do isolamento geográfico. Estudos futuros devem ser realizados para verificar a possível presença do haplótipo recentemente relatado, ac9, em outras áreas do país, além da zona portuária da cidade do Rio de Janeiro / Angiostrongylus cantonensis is respons ible for causing eosinophili c meningoencephalitis in humans and cases have been recorded in various part s of the world including Brazil (ES, PE and SP) . In this study, we report the geneti c variability among Brazilian isolates of A. cantonensis using seque nces of the mitochondrial COI gen e . We identified three Brazilian haplotypes of A. cantonensis , based o n eight known haplotypes (ac1 - ac8) . The Brazilian haplotype ac 5 , was clustered with isolates from Japan and the Brazilian haplotype ac8 ( is olates from R J, SP, PA and PE) formed a distinct clade . It was reported a new Brazilian haplotype , haplotype ac9 , which is closely re lated to haplotype from China (ac6) and Japan (ac7) . Two Brazilian isolates of A. cantonensis , Olinda and Caju (haplotypes ac8 and ac9 , respectively ) reported in this st udy, had their biology and morphology characterized after experimental infection . Significant differences were observed with higher parasite load recovered in the isolates from Caju and a significantly greater number of L1 larvae eliminated in the feces at the beg inning of the patent pe riod . However , when compared to the total larvae eliminated there was no significant difference between the two isolates . The isolates from Caju showed significant difference in the proportion of female and male spec imens (0,64 :1) , but it was not observe d for isolates from Olinda (1,16 :1 ) . The m orphom etric analysis showed that male and female spec imens from Olinda were significantly higher with respect to the analyzed characters when c ompared w it h specimens from Caju. The m orphologic al analysis showed little variation in the level of bifurcations that unite the lateral rays in the right lobe of copulatory bursa , betwe en the two isolates . Genetic variation among isolates supports the hypothesis t hat the appearance of the parasite in Brazil is a result of multiple introd uctions of infected rodents and by the mollusc , Achatina fulica , which contributes to the dispersion . Biological, morphological and morphometric variation between the two haplotype s of A. cantonensis studied , reinforce the observed v ariation by the COI marker and the possible influence of geographical isolation. Future studies should be performed to verify the possible presence of the haplotype recently reported, ac 9, in other areas of the country, beyond the port area of the city of Rio de Janeiro.

A angiostrongilíase abdominal (AA) é uma doença causada pelo nematódeo Angiostrongylus costaricensis, tendo como hospedeiros definitivos roedores silvestres e hospedeiros intermediários moluscos terrestres. No homem, hospedeiro acidental, pode causar infartos intestinais e pseudotumorações, que podem complicar com peritonite e sepse. O diagnóstico definitivo é realizado pelo estudo histopatológico de peças cirúrgicas, ao serem identificadas estruturas parasitárias como vermes, ovos ou larvas. Este processo demanda tempo e experiência do patologista. Até o presente momento não existe tratamento disponível contra o parasito ou para evitar as complicações, apesar de várias tentativas realizadas com o uso de antiparasitários. O presente estudo teve dois objetivos: 1. Testar a reação em cadeia da polimerase (PCR) em bloco parafinado para o diagnóstico de angiostrongilíase abdominal em seres humanos; e 2. Avaliar o efeito da enoxaparina na prevenção de complicações isquêmicas intestinais em camundongos com AA. Para a técnica da PCR utilizamos iniciadores de DNA do Angiostrongylus cantonensis. Blocos parafinados de 4 grupos foram divididos em: (1) casos confirmados (n=20), onde haviam estruturas parasitárias; (2) casos suspeitos (n=20), sendo alvos os granulomas e infiltrado eosinofílico; (3) controle negativo (n=3), proveniente de pacientes operados por adenocarcinomas; e (4) controle de especificidade (n=7), outras parasitoses, representados por estrongiloidíase, enterobíase, ascaridíase e esquistossomose. Para avaliar do tratamento foram utilizados 24 camundongos Swiss infectados com Angiostrongylus costaricensis (10L3/animal), sendo 12 tratados com enoxaparina subcutânea (40mg/kg/dia) e 12 com água destilada (placebo) por 50 dias. Os resultados mostraram que a sensibilidade da PCR foi de 55%, a especificidade de 100% e valor preditivo positivo de 100% para confirmaçãoo da AA. O tratamento dos camundongos com enoxaparina não mostrou diferença com o grupo placebo, em relação a prevenção das lesões tissulares e óbito. Concluindo, a PCR pode tornar-se uma ferramenta promissora para auxiliar na definição diagnóstica da AA, em especial quando o patologista não está familiarizado com esta doença. A enoxaparina na dose profilática não preveniu as lesões e os óbitos causados pelo parasito. / Abdominal angiostrongyliasis (AA) is a disease caused by the nematode Angiostrongylus costaricensis. The parasite has wild rodents as definitive hosts and molusks as intermediate hosts. Incidental contamination in men can provoke bowel infarction and pseudotumoral lesions, clinically manifested as acute abdomen. The diagnosis has been confirmed after histopathological examination of surgical specimens, with the identification of parasitic structures such as worms, eggs and larvae. Diagnostic workup demands high expertise from the pathologist. Hitherto there is no available treatment either for parasite eradication or avoidance of complications, despite several studies addressing the effect of anti-parasitic drugs. The current study aimed: 1. To test polimerase chain reaction (PCR) in formalin-fixed paraffin-embedded (FFPE) tissue for the diagnosis of AA in humans; and 2. To assess the effect of enoxaparin in preventing intestinal ischemic complications in mice with AA. For PCR we used DNA primers from Angiostrongylus cantonensis. Four groups of FFPE intestinal tissue from surgically treated patients were tested with PCR: (1) confirmed cases (n = 20), in which AC structures were present in the target tissue; (2) presumptive cases (n = 20), containing eosinophilic infiltrates and/or granulomatous reaction in the absence of AC structures; (3) negative controls (n = 3), consisting of normal colonic tissue surrounding resected colonic adenocarcinoma; and (4) tissue affected by other parasitosis (n = 7), including strongiloidiasis, ascaridiasis, schistosomiasis, and enterobiasis. In the study with enoxaparin, 24 mice infected with Angiostrongylus costaricensis (10L3/animal) were allocated to receive subcutaneous enoxaparin (40mg/kg/day, n = 12) or sterile water (n = 12) for 50 days. Regarding results, PCR showed sensitivity of 55%, specificity of 100% and positive predictive value of 100% in confirming AA. Treatment with enoxaparin was not superior to placebo in preventing AA related complications or death. In conclusion, the PCR technique might be a promising tool to be used in the diagnostic workup of AA, particularly in the setting of non-experienced pathologists. Enoxaparin in prophylactic doses did not prevent lesions or death in mice suffering from AA.