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Immunomodulatory Response of Mice Splenocytes Induced by RcaL, a Lectin Isolated from Cobia Fish (Rachycentron canadum) Serum

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Abstract

This work reports the isolation of a serum lectin from cobia fish (Rachycentron canadum) named RcaL. Immunomodulatory activity on mice splenocyte experimental cultures through cytotoxic assays and cytokine production were also performed. RcaL was obtained through precipitation with ammonium sulphate and affinity chromatography on a Concanavalin A-Sepharose 4B column. The ammonium sulphate fraction F3 showed the highest specific hemagglutinating activity and was applied to affinity chromatography. The lectin was eluted with methyl-α-d-mannopyranoside. RcaL showed highest affinity for methyl-α-d-mannopyranoside and d-mannose; eluted fractions of RcaL agglutinated rabbit erythrocytes (titre, 128−1) retained 66 % of chromatographed lectin activity, and the obtained purification factor was 1.14. Under reducing conditions, a polypeptide band of 19.2 kDa was revealed in sodium dodecyl sulphate polyacrylamide gel electrophoresis (PAGE). PAGE confirmed RcaL as an acidic protein revealed in a single band. Cytotoxic and immunomodulatory assays with RcaL in mice splenocyte cultures showed that the lectin was not cytotoxic and induced higher interferon gamma and nitric oxide production in splenocyte cultures. Purified RcaL induced preferential Th1 response, suggesting that it acts as an immunomodulatory compound.

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Acknowledgements

The authors express their gratitude to the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) for research grants and fellowship (LCBBC). In addition, the Fundação de Amparo a Ciência e Tecnologia do Estado de Pernambuco (FACEPE) is acknowledged for its financial support. The authors are deeply grateful for the technical assistance of Mrs. Maria Barbosa Reis da Silva.

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Authors and Affiliations

  1. Departamento de Bioquímica, Laboratório de Glicoproteínas, Universidade Federal de Pernambuco/UFPE, Av. Prof. Moraes Rego s/n, Cidade Universitária, CEP: 50670-420, Recife, Pernambuco, Brazil

    Marília Cavalcanti Coriolano, Cynarha Daysy Cardoso da Silva & Luana Cassandra Breitenbach Barroso Coelho

  2. Departamento de Imunologia, Centro de Pesquisas Aggeu Magalhães (CPqAM/FIOCRUZ), Pernambuco, Brazil

    Cristiane Moutinho Lagos de Melo & Valéria Rêgo Alves Pereira

  3. Departamento de Bioquímica, Laboratório de Enzimologia, Universidade Federal de Pernambuco/UFPE, Pernambuco, Brazil

    Ranilson de Souza Bezerra

  4. Departamento de Engenharia de Pesca, Universidade Federal Rural de Pernambuco/UFRPE, Pernambuco, Brazil

    Athiê Jorge Guerra Santos

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  1. Marília Cavalcanti Coriolano
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  2. Cynarha Daysy Cardoso da Silva
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  3. Cristiane Moutinho Lagos de Melo
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Correspondence to Luana Cassandra Breitenbach Barroso Coelho.

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Coriolano, M.C., da Silva, C.D.C., de Melo, C.M.L. et al. Immunomodulatory Response of Mice Splenocytes Induced by RcaL, a Lectin Isolated from Cobia Fish (Rachycentron canadum) Serum. Appl Biochem Biotechnol 168, 1335–1348 (2012). https://doi.org/10.1007/s12010-012-9861-y

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