Inmunogenicidad de la proteína recombinante ASP1R de ancylostoma caninum en un modelo murino.

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Maria Giraldo G Jhon Castaño

Resumen

Objetivo. Construir un plásmido recombinante que exprese la proteína ASP1r de Ancylostoma caninum y evaluar su capacidad inmunogénica en un modelo murino. Materiales y métodos. Se realizó extracción de ARN de parásitos adultos de Ancylostoma caninum, se amplificó por RT –PCR el gen de la proteína ASP1. Este gen fue insertado en el vector pcDNA3. El inserto fue digerido con Bamh1 y EcoR1 y clonado direccionalmente. Posteriormente, se llevó a cabo transformación y selección de las células de E. coli DH5a competentes con el producto de la ligación. Se realizó un tamizaje por PCR confirmando la presencia del gen ASP1. El vector pcDNA3-ASP1 fue administrado vía intraglandular en la parotida e intramuscular en ratones Balb/c. En estos animales se les realizó determinación de anticuerpos en suero y saliva mediante las técnicas de ELISA e inmunohistoquímica. Resultados. Se determinó que el plásmido pcDNA3-ASP1 fue incorporado y expresado células E. coli DH5a. Este plásmido recombinante indujo la producción de anticuerpos Anti-ASP1 específicos en ratones Balb/c. Conclusiones. Se logró demostrar que la utilización de pcDNA3-ASP1 no produjo reacciones desfavorables en ratones Balb/c, además indujo respuesta humoral contra la proteína pcDNA3-ASP1 de excreción/secreción de Ancylostoma caninum en ratones.

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