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Monoclonal antibody generated against Nile tilapia (Oreochromis niloticus) IgT heavy chain using a peptide-based strategy.
Velázquez, Janet; Cruz, Lynn; Pérez-Bernal, Maylin; Valdivia, Onel; Haidar, Arlette; Rodríguez, Alianet; Herrera, Fidel; González, Osmany; Morales, Antonio; Ulloa, Lisbet; Blanco, Reinaldo; Pérez, Joel; Dorta, Dayamí; Luna, Yaramis; Garay, Hilda Elisa; Abreu, David Diago; Ramos, Yassel; Besada, Vladimir; Cabrera, Yeosvany; Estrada, Mario Pablo; Carpio, Yamila.
Afiliación
  • Velázquez J; Animal Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana 10600, Cuba.
  • Cruz L; Animal Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana 10600, Cuba.
  • Pérez-Bernal M; Research and Development Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 83, Sancti Spíritus 60200, Cuba.
  • Valdivia O; Research and Development Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 83, Sancti Spíritus 60200, Cuba.
  • Haidar A; Animal Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana 10600, Cuba.
  • Rodríguez A; Animal Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana 10600, Cuba.
  • Herrera F; Animal Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana 10600, Cuba.
  • González O; Animal Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana 10600, Cuba.
  • Morales A; Animal Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana 10600, Cuba.
  • Ulloa L; Research and Development Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 83, Sancti Spíritus 60200, Cuba.
  • Blanco R; Production Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 83, Sancti Spíritus 60200, Cuba.
  • Pérez J; Production Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 83, Sancti Spíritus 60200, Cuba.
  • Dorta D; Production Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 83, Sancti Spíritus 60200, Cuba.
  • Luna Y; Production Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 83, Sancti Spíritus 60200, Cuba.
  • Garay HE; Peptides Synthesis Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana 10600, Cuba.
  • Abreu DD; Peptides Synthesis Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana 10600, Cuba.
  • Ramos Y; Proteomics Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana, 10600, Cuba.
  • Besada V; Proteomics Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana, 10600, Cuba.
  • Cabrera Y; Research and Development Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 83, Sancti Spíritus 60200, Cuba.
  • Estrada MP; Animal Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana 10600, Cuba.
  • Carpio Y; Animal Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana 10600, Cuba.
Fish Shellfish Immunol Rep ; 4: 100093, 2023 Dec.
Article en En | MEDLINE | ID: mdl-37122444
ABSTRACT
Teleost IgT/Z plays a principal role in the defense mechanisms against infectious agents in the mucosal compartments and in systemic immunity. Previously, Nile tilapia (Oreochromis niloticus) IgT was discovered and characterized at transcription level. In this work, we generated a monoclonal antibody (mAb) that specifically recognized the Nile tilapia IgT. BALB/c mice were immunized with three synthetic peptides conjugated to KLH. The sequences of these peptides derived from the constant region of the Nile tilapia IgT heavy chain. ELISA and Western blotting confirmed the specificity of the polyclonal sera and the culture supernatant from a positive hybridoma clone. We observed immunoreactivity against a recombinant IgT fragment and native IgT in skin mucus. The anti-IgT mAb did not cross-react with purified tilapia IgM. Direct ELISA analysis allowed the quantification of skin mucus IgM and IgT concentrations. Flow cytometry analysis revealed differences in the percentage of IgT+ B cell populations between juveniles and adults in peripheral blood, head kidney and spleen lymphocytes and among the tissues analyzed. For further validation of the anti-IgT mAb utility, a recombinant vaccine candidate against sea lice (TT-P0 Ls) was injected into juvenile tilapia. Direct ELISA results revealed a differential secretion of skin mucus IgT and IgM after immunostimulation. In addition, the percentages of IgT+ B cells were determined at 7 days after booster and ex-vivo stimulation by flow cytometry. This mAb constitutes an important immunological tool to study the biological function and structural characteristics of tilapia IgT.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Fish Shellfish Immunol Rep Año: 2023 Tipo del documento: Article País de afiliación: Cuba
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Fish Shellfish Immunol Rep Año: 2023 Tipo del documento: Article País de afiliación: Cuba