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Fish Shellfish Immunol ; 39(2): 450-7, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24929243

RESUMO

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.


Assuntos
Astacoidea/química , Astacoidea/imunologia , Hemócitos/imunologia , Lectinas/análise , Testes de Aglutinação , Aminoácidos/análise , Animais , Western Blotting , Cromatografia de Afinidade , Eletroforese em Gel Bidimensional , Hemócitos/metabolismo , Hemolinfa/metabolismo , Imuno-Histoquímica , Lectinas/sangue , Lectinas/imunologia , Espectrometria de Massas , Fagocitose/imunologia , Espécies Reativas de Oxigênio/metabolismo , Explosão Respiratória/imunologia , Estatísticas não Paramétricas
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