title: Purification and characterization of a mannose recognition lectin from Oreochromis niloticus (tilapia fish): cytokine production in mice splenocytes.
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.
authors.affiliation: 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.
authors.forename: Cynarha Daysy Cardoso
authors.initials: CD
authors.lastname: da Silva
authors.forename: Marília Cavalcanti
authors.initials: MC
authors.lastname: Coriolano
authors.forename: Mércia Andréa
authors.initials: MA
authors.lastname: da Silva Lino
authors.forename: Cristiane Moutinho Lagos
authors.initials: CM
authors.lastname: de Melo
authors.forename: Ranilson
authors.initials: R
authors.lastname: de Souza Bezerra
authors.forename: Elba Verônica Matoso Maciel
authors.initials: EV
authors.lastname: de Carvalho
authors.forename: Athiê Jorge Guerra
authors.initials: AJ
authors.lastname: Dos Santos
authors.forename: Valéria Rêgo Alves
authors.initials: VR
authors.lastname: Pereira
authors.forename: Luana Cassandra Breitenbach Barroso
authors.initials: LC
authors.lastname: Coelho
mesh_terms: D000818:Animals; D002846:Chromatography, Affinity; D023681:Cichlids / Q000097:blood*; D006384:Hemagglutination / Q000187:drug effects; D007155:Immunologic Factors / Q000097:blood / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D007371:Interferon-gamma / Q000096:biosynthesis*; D008297:Male; D037601:Mannose-Binding Lectin / Q000097:blood / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009573:Nitrites / Q000378:metabolism; D013154:Spleen / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism*
chemical_list: D007155:Immunologic Factors; D037601:Mannose-Binding Lectin; D009573:Nitrites; D007371:Interferon-gamma