Gestation and breastfeeding in schistosomotic mothers differently modulate the immune response of adult offspring to postnatal Schistosoma mansoni infection

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.

Ultrastructural and biochemical detection of biotin and biotinylated polypeptides in Schistosoma mansoni

Biotinylation is proposed for the identification of surface proteins in Schistosoma mansoni using the streptavidin-HRP conjugate for the detection of labeled polypeptides. However, control samples also showed several endogenous biotinylated polypeptides. In an attempt to determine the possibility of nonspecific binding between the streptavidin-HRP conjugate and polypeptides from S. mansoni, the conjugate was blocked with biotinamidecaproate-N-hydroxysuccinimide ester (BcapNHS) before biotin-streptavidin blotting. No bands were detected on the nitrocellulose sheet, demonstrating the specific recognition of biotin by the streptavidin present in the conjugate. Whole cercariae and cercarial bodies and tails showed several endogenous biotinylated polypeptides. The biotin concentration was 13 µg/190,000 cercariae. Adult worms presented less endogenous biotinylated polypeptides than cercariae. These results may be due to changes in the environment from aerobic to anaerobic conditions when cercarial bodies (schistosomula) are transformed into adult worms and a decrease in CO2 production may occur. Cercariae, cercarial bodies and adult male worms were examined by transmission electron microscopy employing an avidin-colloidal gold conjugate for the detection of endogenous biotin. Gold particles were distributed mainly on the muscle fibers, but dispersed granules were observed in the tegument, mitochondria and cytosol. The discovery of endogenous biotin in S. mansoni should be investigated in order to clarify the function of this vitamin in the parasite

Cytokines as determinants of resistance and pathology in human Schistosoma mansoni infection

The role of different cytokines in the peripheral blood mononuclear cell (PBMC) proliferative response and in in vitro granuloma formation was evaluated in a cross-sectional study with patients with the different clinical forms and phases of Schistosoma mansoni infection, as well as a group of individuals "naturally" resistant to infection named normal endemic (NE). The blockage of IL-4 and IL-5 using anti-IL-4 and anti-IL-5 antibodies significantly reduced the PBMC proliferative response to soluble egg (SEA) and adult worm (SWAP) antigens in acute (ACT), chronic intestinal (INT) and hepatosplenic (HS) patients. Similar results were obtained in the in vitro granuloma formation. Blockage of IL-10 had no significant effect on either assay using PBMC from ACT or HS. In contrast, the addition of anti-IL-10 antibodies to PBMC cultures from INT patients significantly increased the proliferative response to SEA and SWAP as well as the in vitro granuloma formation. Interestingly, association of anti-IL-4 and anti-IL-10 antibodies did not increase the PBMC proliferative response of these patients, suggesting that IL-10 may act by modulating IL-4 and IL-5 secretion. Addition of recombinant IL-10 decreased the proliferative response to undetectable levels when PBMC from patients with the different clinical forms were used. Analysis of IFN-g in the supernatants showed that PBMC from INT patients secreted low levels of IFN-g upon antigenic stimulation. In contrast, PBMC from NE secreted high levels of IFN-g. These data suggest that IL-10 is an important cytokine in regulating the immune response and possibly controlling morbidity in human schistosomiasis mansoni, and that the production of IFN-g may be associated with resistance to infection.

Collagen arrangement in hepatic granuloma in mice infected with Schistosoma mansoni: dependence on fiber radiation centers

The collagen structure of isolated and in situ liver granuloma from Swiss Webster mice infected with Schistosoma mansoni was sequentially and three-dimensionally analyzed during different times of infection (early acute, acute, transitional acute-chronic, and chronic phases) by laser scanning confocal microscopy and electron scanning variable vacuum microscopy. The initial granuloma structure is characterized by vascular collagen residues and by anchorage points (or fiber radiation centers), from where collagenous fibers are angularly shed and self-assembled. During the exudative-productive stage, the self-assembly of these fibers minimizes energy and mass through continuous tension and focal compression. The curvature or angles between collagen fibers probably depends on the fibroblastic or myofibroblastic organization of stress fibers. Gradually, the loose unstable lattice of the exudative-productive stage transforms into a highly packed and stable architecture as a result of progressive compactness. The three-dimensional architecture of granulomas provides increased tissue integrity, efficient distribution of soluble compounds and a haptotactic background to the cells.

Sm14 gene expression in different stages of the Schistosoma mansoni life cycle and immunolocalization of the Sm14 protein within the adult worm

Sm14 is a 14-kDa vaccine candidate antigen from Schistosoma mansoni that seems to be involved in cytoplasmic trafficking of fatty acids. Although schistosomes have a high requirement for lipids, they are not able to synthesize fatty acids and sterols de novo. Thus, they must acquire host lipids. In order to determine whether Sm14 is present in different stages of the life cycle of the parasite, we performed RT-PCR. Sm14 mRNA was identified in all stages of the life cycle studied, mainly schistosomulum, adult worm and egg. Additionally, we used a rabbit anti-Sm14 polyclonal antibody in an indirect immunofluorescence assay to localize Sm14 in adult worm sections. The basal lamella of the tegument and the gut epithelium were strongly labeled. These tissues have a high flow of and demand for lipids, a finding that supports the putative role of Sm14 as an intracellular transporter of fatty acids from host cells.

Indirect effects of oral tolerance to ovalbumin interfere with the immune responses triggered by Schistosoma mansoni eggs

The objective of the present study was to investigate whether the injection of a tolerated protein (indirect effects) affects the formation of granulomas around Schistosoma mansoni eggs trapped in the lungs after intravenous (iv) injection into normal (noninfected) C57BL/6 mice (6 animals per group). To induce oral tolerance to chicken egg ovalbumin a 1/5 dilution of egg white in water was offered ad libitum in a drinking bottle for 3 days. Control mice received water. After 7 days, control and experimental animals were injected iv with 2,000 S. mansoni eggs through a tail vein. In some mice of both groups the iv injection of eggs was immediately followed by intraperitoneal (ip) immunization with 10 µg of dinitrophenylated conjugates of ovalbumin (DNP-Ova) emulsified in complete Freund's adjuvant (CFA) or only CFA; 18 days later, mice were bled and killed by ether inhalation. The lungs were fixed in formalin and embedded in paraffin. Serial sections of 5 µm were stained with Giemsa, Gomori's silver reticulin and Sirius red (pH 10.2). Granuloma diameters were measured in histological sections previously stained with Gomori's reticulin. Anti-DNP and anti-soluble egg antigen (SEA) antibodies were analyzed by ELISA. In mice orally tolerant to ovalbumin the concomitant ip injection of DNP-Ova resulted in significantly lower anti-SEA antibodies (ELISA*: 1395 ± 352 in non-tolerant and 462 ± 146 in tolerant mice) and affected granuloma formation around eggs, significantly decreasing granuloma size (area: 22,260 ± 2478 to 12,993 ± 3242 µm²). Active mechanisms triggered by injection of tolerated antigen (ovalbumin) reduce granuloma formation.

Effect of praziquantel administration on hepatic stereology of mice infected with Schistosoma mansoni and fed a low-protein diet

A study was undertaken to investigate the effect of administering praziquantel (PZQ), focusing on the liver stereological findings of malnourished mice infected with Schistosoma mansoni. Thirty female Swiss Webster mice (age: 21 days; weight: 8-14 g) were fed either a low-protein diet (8%) or standard chow (22% protein) for 15 days. Five mice in each group were infected with 50 cercariae each of the BH strain (Brazil). PZQ therapy (80 mg/kg body weight, per day) was started on the 50th day of infection and consisted of daily administration for 5 days. Volume density (hepatocytes, sinusoids and hepatic fibrosis) was determined by stereology using a light microscope. Body weight gain and total serum albumin levels were always lower in undernourished mice. Our stereological study demonstrated that treatment increased both volume density of hepatocytes in mice fed standard chow (47.56%, treated group and 12.06%, control) and low-protein chow (30.98%, treated group and 21.44%, control), and hepatic sinusoids [standard chow (12.52%, treated group and 9.06%, control), low-protein chow (14.42%, treated group and 8.46%, control)], while hepatic fibrosis was reduced [standard chow (39.92%, treated group and 78.88%, control) and low-protein chow (54.60%, treated group and 70.10%, control)]. On the other hand, mice fed low-protein chow decreased density volume of hepatocytes and hepatic fibrosis. In conclusion, our findings indicate that treatment with PZQ ameliorates hepatic schistosomiasis pathology even in mice fed a low-protein diet.

Focos ativos de esquistossomose mansoni no Vale do Ribeira, Estado de São Paulo, Brasil

São apresentados os resultados das investigações sobre os focos de esquistossomose nas localidades de Pedro de Toledo e Itariri, com referência especial aos índices de infecção natural obtidos para a espécie Biomphalaria tenagophila. Êste molusco, transmissor de esquistossomose no Vale do Ribeira já foi assinalado nos seguintes municípios daquela região: Eldorado, Iguape, Itariri, Jacupiranga, Juquiá, Miracatu, Pariqueraçu, Pedro de Toledo, Registro e Sete Barras. Os índices cercáricos de 68,1%, 75,0% e 81,3% obtidos em alguns lotes de B. tenagophila, os mais altos já verificados para esta espécie, vem mais uma vez comprovar sua grande suscetibilidade à infecção pelo Schistosoma mansoni.

Colonização de Pomacea haustrum (Reeve, 1856) em localidade com esquistossomose mansoni: Baldim, MG (Brasil). (Prosobranchia, Pilidae)

Na localidade de Baldim, MG, Brasil, foram introduzidos, em agosto de 1972, 5.421 exemplares de Pomacea haustrum (Prosobranchia, Pilidae) em 5 córregos e 2 valas, nos quais predominavam Biomphalaria glabrata (Say, 1818) e, secundariamente, B. straminea (Dunker, 1848). Entre 1968 e 1971, os índices de infecção da espécie B. glabrata por Schistosoma mansoni oscilaram de 2,1% a 11,9%. Em nenhum momento foram capturados B. straminea liberando cercárias daquele trematódeo. Após a introdução do pilídeo, apenas uma única vez detectou-se 2 (0,8%) B. glabrata positivas. Observou-se decréscimo populacional de planorbineos e aumento de densidade de pomácea até 20,0 e 121,6 exemplares/m² em córregos e valas, respectivamente. A estimativa da densidade de F. haustrum foi feita através do método dos "quadrats". Foram coletados, de junho de 1968 a julho de 1972, 65,2% (1.526) dos planorbíneos. Porém, após a introdução do predador-competidor, foram registrados os seguintes dados: 1976, 15% (352); em 1977, 16,1% (377) e, em 1978, apenas 3,7% (87) do total dos exemplares capturados. As pomáceas, transferidas do ambiente lenítico (Sete Lagoas, MG), adaptaram-se às coleções lóticas de Baldim e foram capazes de substituir as populações originais de B. glabrata em vários biótopos, ou tornaram-se, pelo menos, dominantes, sem danos visíveis para os novos ecossistemas. Acredita-se que em outras situações análogas, Pomacea haustrum (Reeve, 1956) - e, por extensão, P. lineata (Spix, 1827), P. canaliculata (Lamark, 1822) e outras do mesmo táxon - poderão ser utilizadas, com sucesso, no controle biológico dos hospedeiros intermediários de Schistosoma mansoni.

Ação inibidora de termofosfato magnesiano sobre a fecundidade de planorbídeos e seu possível significado no controle da esquistossomose mansoni

A eficácia de termofosfato magnesiano (fertilizante e corretivo do solo) na alimentação de planorbídeos, em laboratório, foi testada ao longo de cinco semanas, verificando-se o crescimento da concha de populações experimentais de Bimphalaria glabrata (Say, 1818) e Helisoma duryi (Wetherby, 1879). Reversamente observou-se que aquele produto mineral bloqueou a oviposição de ambas as espécies, a partir da segunda semana do experimento, quando proporcionado, isoladamente ou associado, a latoso ou a folhas de alface. Em conseqüência, supõe-se que sua ampla utilização nas novas áreas de cultivo, obtidas através de métodos modernos de irrigação, poderá, adicionalmente - ao ser carreado pelas chuvas, para ecossistemas lênticos ou lóticos - reduzir as populações pioneiras de diferentes espécies de Bimphalaria constituindo-se útil meio de controle da esquistossomose mansoni, em zonas rurais. Ademais, poderá ser eficaz no controle dos hospedeiros intermediários de Schistosoma mansoni que, usualmente, se estabelecem em lagoas de criação de peixes - cuja multiplicação, em áreas tropicais, vem sendo incentivada, quer como atividade econômica alternativa para o pequeno proprietário rural quer como exigência de necessidade nutricional das comunidades humanas mais pobres. Supõe-se, ainda, que o material testado poderá controlar populações de limneídeos, hospedeiros intermediários de Fasciola hepatica. São sugeridos testes de campo que possam efetivamente comprovar a hipótese levantada, e também estudos destinados ao conhecimento do mecanismo de ação do elemento magnésio no bloqueio da fertilidade de moluscos. Salienta-se a necessidade de pesquisas especiais sobre o efeito de diferentes concentrações de magnésio em ovos e formas juvenís de planorbídeos, bem como sobre miracídios e cercárias (intra-caramujos e livre-natantes) de trematódeos.

Estudo clínico-epidemiológico da esquistossomose mansoni em escolares da Ilha, município de Arcos, MG (Brasil), 1983

Foi realizado um estudo clínico-epidemiológico da esquistossomose em escolares (6 - 14 anos) da Ilha, em Minas Gerais (Brasil). Foram feitos exame parasitológico de fezes pelo método de KATO-KATZ e exame clínico em, respectivamente, 86,7 e 85,4% da população escolar. Foi realizado levantamento sócio-econômico e foram pesquisados os sinais e sintomas apresentados pelos pacientes e seus contatos com águas naturais. O índice de infecção pelo Schistosoma mansoni foi 32,7%, predominaram as baixas contagens de ovos nas fezes (89,0% dos pacientes eliminavam menos de 500 ovos/grama de fezes) e o índide de esplenomegalia foi 7,7%. Os pacientes positivos (com ovos de S. mansoni nas fezes) foram comparados aos negativos (sem ovos nas fezes e com intradermoreação negativa), verificando-se que a infecção pelo S. mansoni era significativamente mais freqüente entre os trabalhadores rurais, entre os que residiam em casas de pior qualidade e entre aqueles cujos chefes de família eram analfabetos. Constituíram os maiores fatores de risco para a infecção pelo S. mansoni nesta área: a) ter contatos com águas naturais para trabalhar na lavoura (Odds Ratio = 18,08); b) ter contatos diários com águas naturais (OR = 13,82) e c) ter contatos com águas naturais para pescar, nadar e/ou brincar (OR = 7,75 e 5,51, respectivamente). Os autores levantam a hipótese de que a transmissão da esquistossomose nesta localidade não ocorre no peridomicílio, mas sim nas lagoas próximas à Ilha e nas plantações agrícolas, provavelmente nas culturas de arroz de várzea.

Situação atual da esquistossomose mansoni no Lago da Pampulha, Belo Horizonte, MG, Brasil

Descreve-se a atual distribuição dos planorbídeos hospedeiros intermediários do Schistosoma mansoni no Lago da Pampulha, Belo Horizonte, MG (Brasil) e a rápida dispersão de Biomphalaria tenagophila por todo o perímetro lacustre. Relata-se o primeiro encontro de B. tenagophila naturalmente infectada por S. mansoni, naquela coleção hídrica. Discute-se, a importância epidemiológica destas observações e a possibilidade da B. tenagophila vir a substituir a primitiva população de B. glabrata no Lago da Pampulha.

Primeiros casos autóctones de esquistossomose mansoni em região do Noroeste do Estado de Minas Gerais (Brasil)

São relatados os primeiros casos autóctones de esquistossomose mansoni, originários da cidade de Paracatu, Noroeste do Estado de Minas Gerais, Brasil, região até o momento considerrada indene para esta parasitose. Trata-se de oito pacientes com idade entre 12 e 14 anos, e que nunca haviam se ausentado da localidade. Os levantamentos malacológicos proporcionaram a coleta de 11.471 exemplares de Biomphalaria straminea, todos negativos para Schistosoma mansoni. Foram sugeridas medidas que, se colocadas em prática, poderiam controlar a parasitose na cidade.

The epidemiology and control of schistosomiasis mansoni where Biomphalaria tenagophila is the snail host

The epidemiology and control of schistosomiasis mansoni in the Municipality of Pedro de Toledo (State of S. Paulo, Brazil) since 1980, has been studied. In 1980 the prevalence evaluated by stool exams (Kato-Katz method) was 22.8% and no statistical difference at 5.0% level was observed between rural and urban zones. The intensity of infection was low (58.5 eggs/g of faeces); the highest prevalence and intensity of infection rates were observed within the group of from 5 to 29 years of age, respectively. The transmission of schistosomiasis usually occurred during leisure time. The majority of the carriers of the parasite were asymptomatic. Of the B. tenagophila examined only 0.4% were found to be infected. The control programme has been intensified from 1981 on resulting in a sharp decrease in the prevalence from 22.8% in 1980 to 6% at the present time. This result shows that, in spite of the control programme there is a residual human prevalence. A beginning has been made on the investigation into the possible causes of this residual prevalence (6.0% was maintained through out 1987).