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Purification and Characterization of a Mannose Recognition Lectin from Oreochromis niloticus (Tilapia Fish): Cytokine Production in Mice Splenocytes

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Abstract

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.

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Acknowledgments

The authors express their gratitude to the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) for research grants, fellowships (RSB and LCBBC), and a postdoctoral fellowship (EVMMC). Also, the Fundação de Amparo a Ciência e Tecnologia do Estado de Pernambuco (FACEPE) is acknowledged for its financial support and the Estação de Aqüicultura Continental Johei Koike, Departamento de Pesca e Aqüicultura (UFRPE) for fish donation. The authors are deeply grateful for the technical assistance of Mrs. Maria Barbosa Reis da Silva and Lucas Ferreira da Rocha.

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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

    Cynarha Daysy Cardoso da Silva, Marília Cavalcanti Coriolano, Mércia Andréa da Silva Lino, Elba Verônica Matoso Maciel de Carvalho & Luana Cassandra Breitenbach Barroso Coelho

  2. Departamento de Imunologia, Centro de Pesquisas Aggeu Magalhães–CPqAM/FIOCRUZ, Recife, Pernambuco, Brazil

    Cristiane Moutinho Lagos de Melo & Valéria Rêgo Alves Pereira

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

    Ranilson de Souza Bezerra

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

    Athiê Jorge Guerra dos Santos

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  1. Cynarha Daysy Cardoso da Silva
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  2. Marília Cavalcanti Coriolano
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  3. Mércia Andréa da Silva Lino
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  6. Elba Verônica Matoso Maciel de Carvalho
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  7. Athiê Jorge Guerra dos Santos
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  8. Valéria Rêgo Alves Pereira
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  9. Luana Cassandra Breitenbach Barroso Coelho
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Correspondence to Luana Cassandra Breitenbach Barroso Coelho.

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da Silva, C.D.C., Coriolano, M.C., da Silva Lino, M.A. et al. Purification and Characterization of a Mannose Recognition Lectin from Oreochromis niloticus (Tilapia Fish): Cytokine Production in Mice Splenocytes. Appl Biochem Biotechnol 166, 424–435 (2012). https://doi.org/10.1007/s12010-011-9438-1

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  • DOI: https://doi.org/10.1007/s12010-011-9438-1

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