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Fish Shellfish Immunol ; 39(2): 450-7, 2014 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-24929243

RESUMEN

Lectins participate in the immune mechanisms of crustaceans. They have been considered as humoral receptors for pathogen-associated molecular patterns; however, some reports suggest that lectins could regulate crustacean cellular functions. In the present study, we purified and characterized a serum lectin (CqL) from the hemolymph of Cherax quadricarinatus by affinity chromatography and determined its participation in the regulation of hemocytes' oxidative burst. CqL is a 290-kDa lectin in native form, constituted by 108, 80, and 29-kDa subunits. It is mainly composed of glycine, alanine, and a minor proportion of methionine and histidine. It showed no carbohydrates in its structure. CqL is composed of several isoforms, as determined by 2D-electrophoresis, and shows no homology with any crustacean protein as determined by Lc/Ms mass spectrometry. CqL agglutinated mainly rat and rabbit erythrocytes and showed a broad specificity for monosaccharides such as galactose, glucose, and sialic acid, as well as for glycoproteins, such as porcine stomach and bovine submaxillary mucin and fetuin. It is a Mn(2+)-dependent lectin. CqL recognized 8% of crayfish granular hemocytes and increased 4.2-fold the production of hemocytes' superoxide anion in vitro assays when compared with non-treated hemocytes. This effect showed the same specificity for carbohydrates as hemagglutination; moreover, superoxide dismutase and diphenyleneiodonium chloride were effective inhibitors of CqL oxidative-activation. The CqL homoreceptor is a 120-kDa glycoprotein identified in the hemocytes lysate. Our results suggest that CqL participates actively in the regulation of the generation of superoxide anions in hemocytes using NADPH-dependent mechanisms.


Asunto(s)
Astacoidea/química , Astacoidea/inmunología , Hemocitos/inmunología , Lectinas/análisis , Pruebas de Aglutinación , Aminoácidos/análisis , Animales , Western Blotting , Cromatografía de Afinidad , Electroforesis en Gel Bidimensional , Hemocitos/metabolismo , Hemolinfa/metabolismo , Inmunohistoquímica , Lectinas/sangre , Lectinas/inmunología , Espectrometría de Masas , Fagocitosis/inmunología , Especies Reactivas de Oxígeno/metabolismo , Estallido Respiratorio/inmunología , Estadísticas no Paramétricas
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