RESUMO
Skin schistosomula can be prepared by collecting them after isolated mouse skin have been penetrated by cercariae in vitro. The schistosomula can also migrate out of isolated mouse skin penetrated by cercariae in vitro and from mouse skin penetrated by cercariae in vivo. Schistosomula can also be produced from cercariae applied through a syringe or in a vortex. When certain surface properties of the different forms of schistosomula were compared, those migrating from mouse skin penetrated by cercariae in vivo or in vitro had greatly increased permeability to membrane impermeant molecules such as Lucifer yellow and high molecular weight dextrans. These migrating forms also possessed surfaces which showed greatly enhanced uptake into internal membrane vesicles of the dye FM 143, a marker for endocytosis. This greatly enhanced activity and permeability of the surfaces of tissue migrating schistosomula is likely to be of great importance in the adaptation to the new host.
Assuntos
Schistosoma mansoni/fisiologia , Esquistossomose mansoni/parasitologia , Animais , Corantes Fluorescentes , Isoquinolinas/química , Camundongos , Movimento , Permeabilidade , Esquistossomose mansoni/patologia , Pele/parasitologia , Pele/patologiaRESUMO
Introduction The diagnosis of schistosomiasis mansoni on early stages of infection is important to prevent late morbidity. A simple, cheap, sensitive and specific assay for routine diagnosis of schistosome infection based on the detection of specific IgG for schistosomula tegument antigens (ELISA-SmTeg) was developed by our group. Methods We describe here an acute outbreak involving a travel group of 80 individuals from a non-endemic area of the State of Minas Gerais, Brazil. These individuals were in contact with a freshwater pool where Biomphalaria glabrata was found. Results obtained from our new methodology were compared to IgG antibody titers against soluble worm antigenic preparation (SWAP) by ELISA and, also to parasitological examination, nuclear magnetic resonance and clinical findings. Results ELISA-SmTeg was capable of detecting 64 positive cases among the 80 individuals participating at the survey with a positivity ratio of 80% and a higher sensitivity than ELISA-SWAP that was only sensitive for 56% of positive cases. Besides, a significant correlation was found for the severity of the infection and the specific IgG titers against SmTeg. Conclusions Our data showed that ELISA-SmTeg might serve as the initial diagnostic tool for acute stages of the infection in community-based helminth control programs or for the surveillance of individuals from non-endemic areas. .
Assuntos
Animais , Humanos , Anticorpos Anti-Helmínticos/sangue , Antígenos de Helmintos , Surtos de Doenças , Imunoglobulina G , Schistosoma mansoni/imunologia , Esquistossomose mansoni/diagnóstico , Viagem , Doença Aguda , Brasil/epidemiologia , Ensaio de Imunoadsorção Enzimática , Fezes/parasitologia , Espectroscopia de Ressonância Magnética , Contagem de Ovos de Parasitas , Sensibilidade e Especificidade , Esquistossomose mansoni/epidemiologiaRESUMO
If Schistosoma mansoni infection could be detected in its early stages, especially before the egg deposition in the host tissues, the development of severe pathologic lesions could be efficiently prevented. We therefore developed an indirect enzyme-linked immunosorbent assay based on the detection of specific IgG against schistosomula antigens (ELISA-SmTeg). The assay was applied in sera samples from non-infected and infected mice collected seven and 15 days post-infection. The results were compared to the number of adult worms obtained by perfusion of the murine hepatic system 50 days post-infection. The sensitivity and specificity of the ELISA-SmTeg were 100% (p = 0.0032 and 0.0048 respectively for seven and 15 days of infection) with a cutoff value of 0.15 (p = 0.0002). Our findings show a novel low-cost serological assay using antigens which are easy to obtain, which was able to detect all the infected mice as early as seven days post-infection.
A detecção da infecção pelo helminto Schistosoma mansoni quando realizada nas fases iniciais, especialmente antes da oviposição nos tecidos do hospedeiro, pode impedir de forma eficiente o desenvolvimento de graves lesões patológicas. Baseado nisto, foi desenvolvido um ensaio imunoenzimático indireto para detecção de anticorpos IgG específicos contra antígenos de esquistossômulos (ELISA-SmTeg). Este ensaio foi aplicado em amostras sorológicas de camundongos não infectados, da mesma forma que de camundongos recentemente infectados, após sete e 15 dias de infecção. Os resultados foram comparados com o número de vermes adultos obtidos por perfusão do sistema hepático murino 50 dias pós-infecção. A sensibilidade e a especificidade do novo método, denominado ELISA-SmTeg, foram de 100% (p = 0,0032, 0,0048, respectivamente, durante sete e 15 dias de infecção) com um valor de corte de 0,15 (p = 0,0002). Nossos resultados mostraram que um ensaio de baixo custo, que utiliza antígenos de fácil obtenção, é capaz de discriminar a esquistossomose mansoni em modelo experimental de forma precoce, incluindo sete dias pós-infecção.
Assuntos
Animais , Feminino , Camundongos , Anticorpos Anti-Helmínticos/sangue , Antígenos de Helmintos/imunologia , Imunoglobulina G/sangue , Schistosoma mansoni/imunologia , Esquistossomose mansoni/diagnóstico , Anticorpos Anti-Helmínticos/imunologia , Diagnóstico Precoce , Ensaio de Imunoadsorção Enzimática/métodos , Imunoglobulina G/imunologia , Contagem de Ovos de Parasitas , Sensibilidade e EspecificidadeRESUMO
The present investigation involves a reevaluation of previous results obtained after experimental infection of Swiss Webster mice with cercariae and schistosomula of the Schistosoma mansoni LE strain maintained under laboratory conditions. Three experimental groups of mice were considered: the animals of the first group were percutaneously (ring method) infected with cercariae, those of the second were subcutaneously inoculated with cercariae and the mice of the third were inoculated by the same route with schistosomula transformed in vitro. The data obtained so far indicated that the most effective method of infection is the subcutaneous injection with schistosomula, with a mean adult worm burden recovery of 54.1% when compared to the abdominal percutaneous and subcutaneous routes of infection with cercariae, in which the values were 36.7% and 32.4%, respectively. This suggests that, in experimental infections of SW mice with a LE S. mansoni strain, the skin is to be considered an effective attrition site in the percutaneous route, whereas in the case of inoculation with cercariae, a small amount of larvae fails to be transformed into viable schistosomula, possibly due to skin phase avoidance. A brief discussion about attrition sites and elimination of larval S. mansoni worms in mice is presented.
Os objetivos da presente investigação foram os de reavaliar resultados anteriores obtidos através de infecções experimentais de camundongos SW com cercarias e esquistossômulos da cepa LE de Schistosoma mansoni mantida em laboratório. Três grupos de camundongos foram considerados: animais do primeiro grupo foram infectados com cercarias pela via percutânea (método do anel), os do segundo inoculados pela via subcutânea com cercarias e os do terceiro inoculados pela mesma via com esquistossômulos obtidos "in vitro". Os dados obtidos mostraram que a via de infecção mais eficiente é a injeção subcutânea de esquistossômulos transformados "in vitro", com média de recuperação de vermes adultos de 54.1%, quando comparada às vias percutânea abdominal e subcutânea, com médias de 36,7% e 32,4%, respectivamente, sugerindo que em infecções experimentais de camundongos SW com S. mansoni da cepa LE, a pele pode ser considerada um eficaz sítio de atrito na via percutânea, enquanto no caso da inoculação de cercarias, um pequeno número de larvas não se transforma em esquistossômulos viáveis, talvez pelo fato de a barreira da pele haver sido evitada. Breves considerações a respeito dos sítios de atrito e eliminação de formas larvares do S. mansoni em camundongos, são apresentadas.