Desarrollo de biomodelos para la evaluación de la inmunidad secretora contra M. tuberculosis en ratones Balb/c.

Development of Biomodels for the evaluation of the secretory immunity against M tuberculosis in Balb/c mice.

Annette León1, Armando Acosta1, María Elena Sarmiento1, Pedro Estévez1, Máximo Martínez1, María
Elena Pérez1, Gustavo Falero1, Mildrey Fariñas1, Juan Francisco Infante1, Juraj Ivanyi2, Gustavo Sierra1.

RESUMEN

Poco se ha estudiado acerca del papel de los anticuerpos específicos, presentes en las secreciones del aparato respiratorio, en la defensa contra patógenos intracelulares, como es el caso de las micobacterias causantes de la tuberculosis en el hombre: Micobacterium tuberculosis, bovis y africanum. Con el objetivo de desarrollar modelos adecuados para evaluar el posible papel de la inmunidad secretoria en la defensa contra la tuberculosis, se desarrollaron dos modelos animales con la utilización de un anticuerpo monoclonal IgA dirigido contra la proteína de 16 kD de M. tuberculosis y M. bovis. En el primer modelo se inocularon ratones Balb/c, por vía subcutánea al nivel de la nuca, con diferentes cantidades de células del hibridoma TBA61, productor de la IgA específica. En un segundo modelo, se inoculó por vía intraperitoneal líquido ascítico correspondiente a este hibridoma obtenido en ratón. En ambos casos se determinó, a diferentes tiempos, la concentración del monoclonal en saliva y sólo en suero para el segundo. En los dos modelos se demostró el paso del monoclonal a la saliva, donde alcanzó la máxima concentración: a los 21 días en los animales inoculados con el hibridoma, y a las 2 horas en saliva y suero en los animales inoculados con el líquido ascítico. Se sugiere, por su sencillez y mayor inocuidad, el uso del segundo modelo para la realización de estudios de reto por vía mucosal.

Palabras claves: Tuberculosis, inmunidad de mucosa, anticuerpo, hibridoma, IgA.

ABSTRACT

The role of specific antibodies found in the respiratory tract secretion, in the defense against intracellular pathogens like mycobacterium causing tuberculosis in man (M. tuberculosis, M. bovis, M. africanum), has not been studied deeply enough. With the purpose of developing suitable experimental models aimed at the assessment of the secretory immunity role in the defense against tuberculosis, two experimental models were developed using IgA monoclonal antibodies directed against the M. tuberculosis and M. bovis. 16 kDa protein. In the first model Balb/c mice were subcutaneously inoculated in the neck with different amounts of TBA61 hibridome cells, which produce the specific IgA. In a second model, this hibridome’s ascitic fluid, obtained from mice, was intraperitoneally inoculated. In both cases, we determined, in different moments, the monoclonal concentration in saliva and only in the second one; we determined the concentration in serum. In the two ]]> models we demonstrated the monoclonal’s pass to the saliva, reaching its maximum concentration at 21 days in the animals inoculated with the hibridome and at two hours in saliva and serum in the animals inoculated with ascitic fluid. Due to its simplicity and innocuousness, we recommend the use of the second model for challenging experiments through the mucosal way.

Key words: Tuberculosis, mucosal immunity, Antibodies, hibridome, IgA

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