Purification of a cathepsin L-like proteinase secreted by adult Fasciola hepatica

Smith, Amanda M.; Dowd, Andrew; McGonigle, Sharon; Keegan, Paul S.; Brennan, Gerard; Trudgett, Alan; Dalton, John P.

doi:10.1016/0166-6851(93)90171-s

Cited by 136 publications

(77 citation statements)

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“…Indeed, it has been demonstrated that F. h e p a tic a shows perm anent turnover of its surface antigens, inducing the formation o f immune com plex pre cipitates on the juvenile fluke surface (Hanna, 1980;Sandeman & Howell, 1980;Glauert et al, 1985 ;Tkalevic et al, 1996) which prevents effector cells adhering to juvenile flukes. F. h e p a tic a excretorysecretory products (FhESP) contain proteolytic enzymes such as cysteine proteases (cathepsin L-like) (Chapman & Mitchell, 1982;Smith et al, 1993a;Smith et al, 1993b) w hich cleave in vitro immunoglobulins and inhibit the in vitro adherence o f eosinophils to juve nile flukes in the presence o f immune serum (Carmona et al, 1993). It has also been observed that flukes migrating in the hepatic parenchyma are covered by IgM (Chauvin & Boulard, 1996), w hen eosinophils do not possess a Fcp-receptor.…”

Section: Introductionmentioning

confidence: 99%

IFNγ and IL-10 production by hepatic lymph node and peripheral blood lymphocytes inFasciola hepaticainfected sheep

1998

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Section: Introductionmentioning

confidence: 99%

IFNγ and IL-10 production by hepatic lymph node and peripheral blood lymphocytes inFasciola hepaticainfected sheep

1998

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“…Several functions have been suggested for these proteinases, including tissue invasion, feeding and immune evasion [3,32]. In the present work, two cysteine proteinases of 28 and 34 kDa were purified, by means of preparative electrophoresis and electroelution in polyacrylamide gels.…”

Section: Discussionmentioning

confidence: 96%

“…There was no significant correlation between anti-P28 or anti-ESP IgG levels and fluke burdens and no differences could be detected between primarily and secondarily infected goats. This similarity may be explained by the fact that the ESP of adult F. hepatica included cysteine proteinases [32], secreted or excreted from the tegument or the gut of the flukes [2].…”

Section: Discussionmentioning

confidence: 99%

Humoral response (IgG) of goats experimentally infected with Fasciola hepatica against cysteine proteinases of adult fluke

et al. 2003

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-The use of cysteine proteinases from Fasciola hepatica adult flukes for the serodiagnosis of caprine fasciolosis by means of an indirect ELISA test was studied. Two proteolytic fractions from adult fluke homogenates, with apparent molecular weights of 28 and 34 kDa (P28 and P34 respectively), were characterised as cysteine proteinases using azocasein assays and gelatin gel analysis. Both P28 and P34 fractions were electroluted and used as antigens in two different indirect ELISA tests. Serum IgG levels against P28 and P34 in goats given an experimental primary infection with 200 metacercariae or in goats given two experimental infections with 200 metacercariae were determined and compared with those observed in an uninfected control group. ELISA tests using both cysteine proteases showed a rapid and consistent detection of specific IgG in all experimentally infected goats. The IgG response to P28 was the first to be detected as early as 2-3 weeks post-infection and remained elevated throughout the experiment. The response to P34 was detected later (4-6 wpi) and disappeared in some animals at 18 wpi, while flukes were still present in the bile ducts. No significant differences were observed between the anti-P28 and anti-P34 IgG responses between animals receiving a primary or a challenge infection. The results of our study, although preliminary, are promising since the P28 ELISA described here may be a reliable method for the immunodiagnosis of F. hepatica infection in goats.Fasciola hepatica / cysteine proteinases / IgG / ELISA / goat

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“…Las fracciones de pesos moleculares aproximados 27 KDa y 25-26 KDa son de naturaleza proteinasa como catepsina L-like, según el análisis de las secuencias N-terminal, se trata de una de las proteínas más importantes excretadas y secretadas por F. hepatica in vitro y son las moléculas predominantes en un 80%, también se pudo demostrar por microscopía que se encuentran en vesículas a nivel de las células epiteliales del intestino de F. hepatica (13,14) . Si solo nos basaríamos en el peso molecular de la fracción purificada en este trabajo, se podría decir que se trata de la misma proteinasa que tiene mucha importancia en el inmunodiagnóstico de la fascioliasis humana; para demostrar esto es recomendable realizar un trabajo de caracterización molecular y bioquímica de esta fracción; ya que esta proteinasa, catepsina L-like, se usa en otros países en pruebas de ELISA y Western Blot, obteniéndose buenos resultados y que ahora podrían ser utilizados como una proteína de importancia inmunodiagnóstica (15)(16)(17) .…”

Section: Discussionunclassified

Purificación de la fracción antigénica 27-28 KDA a partir del antígeno metabólico secretado excretado de Fasciola hepatica

Antitupa

Quispe

Mayo

et al. 2014

Rev Peru Med Exp Salud Publica

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Citar como: Antitupa I, Quispe W, Mayo J, Valverde F, Sanchez E. Purificación de la fracción antigénica 27-28 KDa a partir del antígeno metabólico secretadoexcretado de Fasciola hepatica. Rev Peru Med Exp Salud Publica. 2014;31(2) RESUMENEn el presente estudio, las fracciones antigénicas de 27-28 KDa de Fasciola hepatica fueron purificadas por cromatografía de exclusión molecular para su aplicación en el diagnóstico de la fascioliasis humana. Se obtuvieron antígenos de excreción y secreción a partir de fasciolas adultas vivas obtenida de hígado de ovino y bovino, y cultivados en medio mínimo esencial. La reactividad y eficacia del antígeno purificado fueron evaluadas por la prueba de inmunoblot empleando cuatro sueros con fascioliasis humana; cuatro sueros con otras parasitosis, y dos sueros negativos. Se concluye que las fracciones antigénicas purificadas no presentan reacción cruzada con otras parasitosis, por inmunoblot, por lo que se considera a las proteínas purificadas como potenciales candidatas a ser utilizadas para el diagnóstico de fascioliasis humana.Palabras clave: Fasciola hepatica; Purificación; Cromatografía (fuente: DeCS BIREME). PURIFICATION OF ANTIGENIC FRACTION 27-28 kDa from the metabolic ANTIGEN FROM METABOLIC secreted-excreted from Fasciola hepatica ABSTRACTAntigenic fractions of 27-28 kDa from Fasciola hepatica were purified by size-exclusion chromatography for use in the diagnosis of human fasciolosis. Excretion and secretion antigens were obtained from living adult flukes collected from sheep and cattle liver, and cultured in minimum essential medium. The reactivity of the purified antigen and efficacy were assessed by immunoblot test using four sera with human fascioliasis; four sera with other parasites, and two negative sera. We conclude that the purified antigenic fractions do not cross-react with other parasites by immunoblot. Therefore, purified proteins are considered as potential candidates to be used for the diagnosis of human fascioliasis.

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Purification of a cathepsin L-like proteinase secreted by adult Fasciola hepatica

Cited by 136 publications

References 14 publications

IFNγ and IL-10 production by hepatic lymph node and peripheral blood lymphocytes inFasciola hepaticainfected sheep

IFNγ and IL-10 production by hepatic lymph node and peripheral blood lymphocytes inFasciola hepaticainfected sheep

Humoral response (IgG) of goats experimentally infected with Fasciola hepatica against cysteine proteinases of adult fluke

Purificación de la fracción antigénica 27-28 KDA a partir del antígeno metabólico secretado excretado de Fasciola hepatica

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