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Appl Biochem Biotechnol ; 166(2): 424-35, 2012 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-22081327

RESUMEN

The aim of this work was to purify and partially characterize a mannose recognition lectin from Nile tilapia (Oreochromis niloticus) serum, named OniL. OniL was isolated through precipitation with ammonium sulfate and affinity chromatography (Concanavalin A-Sepharose 4B). In addition, we evaluated carbohydrate specificity, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) profiles, and in vitro immunomodulatory activity on mice splenocyte experimental cultures through cytotoxic assays and cytokine production. The ammonium sulfate fraction F2 showed the highest specific hemagglutinating activity (331) and was applied to affinity matrix. Adsorbed proteins (OniL) were eluted with methyl-α-D: -mannopyranoside. OniL, a 17-kDa protein by SDS-PAGE constituted by subunits of 11 and 6.6 kDa, showed highest affinity for methyl-α-D: -mannopyranoside and D: -mannose. Immunological assays, in vitro, showed that OniL did not show cytotoxicity against splenocytes, induced higher IFN-γ production and lower IL-10 as well as nitrite release. In conclusion, OniL lectin was successfully purified and showed a preferential Th1 response in mice splenocytes.


Asunto(s)
Cíclidos/sangre , Interferón gamma/biosíntesis , Lectina de Unión a Manosa/aislamiento & purificación , Lectina de Unión a Manosa/farmacología , Bazo/efectos de los fármacos , Bazo/metabolismo , Animales , Cromatografía de Afinidad , Hemaglutinación/efectos de los fármacos , Factores Inmunológicos/sangre , Factores Inmunológicos/aislamiento & purificación , Factores Inmunológicos/metabolismo , Factores Inmunológicos/farmacología , Masculino , Lectina de Unión a Manosa/sangre , Lectina de Unión a Manosa/metabolismo , Ratones , Ratones Endogámicos BALB C , Nitritos/metabolismo , Bazo/inmunología
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