Monoclonal antibody generated against Nile tilapia (Oreochromis niloticus) IgT heavy chain using a peptide-based strategy

Janet Velázquez; Lynn Cruz; Maylin Pérez-Bernal; Onel Valdivia; Arlette Haidar; Alianet Rodríguez; Fidel Herrera; Osmany González; Antonio Morales; Lisbet Ulloa; Reinaldo Blanco; Joel Pérez; Dayamí Dorta; Yaramis Luna; Hilda Elisa Garay; David Diago Abreu; Yassel Ramos; Vladimir Besada; Yeosvany Cabrera; Mario Pablo Estrada; Yamila Carpio

Fish and Shellfish Immunology Reports (Dec 2023)

Monoclonal antibody generated against Nile tilapia (Oreochromis niloticus) IgT heavy chain using a peptide-based strategy

Janet Velázquez,
Lynn Cruz,
Maylin Pérez-Bernal,
Onel Valdivia,
Arlette Haidar,
Alianet Rodríguez,
Fidel Herrera,
Osmany González,
Antonio Morales,
Lisbet Ulloa,
Reinaldo Blanco,
Joel Pérez,
Dayamí Dorta,
Yaramis Luna,
Hilda Elisa Garay,
David Diago Abreu,
Yassel Ramos,
Vladimir Besada,
Yeosvany Cabrera,
Mario Pablo Estrada,
Yamila Carpio

Affiliations

Janet Velázquez: Animal Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana 10600, Cuba
Lynn Cruz: Animal Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana 10600, Cuba
Maylin Pérez-Bernal: Research and Development Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 83, Sancti Spíritus 60200, Cuba
Onel Valdivia: Research and Development Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 83, Sancti Spíritus 60200, Cuba
Arlette Haidar: Animal Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana 10600, Cuba
Alianet Rodríguez: Animal Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana 10600, Cuba
Fidel Herrera: Animal Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana 10600, Cuba
Osmany González: Animal Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana 10600, Cuba
Antonio Morales: Animal Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana 10600, Cuba
Lisbet Ulloa: Research and Development Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 83, Sancti Spíritus 60200, Cuba
Reinaldo Blanco: Production Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 83, Sancti Spíritus 60200, Cuba
Joel Pérez: Production Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 83, Sancti Spíritus 60200, Cuba
Dayamí Dorta: Production Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 83, Sancti Spíritus 60200, Cuba
Yaramis Luna: Production Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 83, Sancti Spíritus 60200, Cuba
Hilda Elisa Garay: Peptides Synthesis Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana 10600, Cuba
David Diago Abreu: Peptides Synthesis Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana 10600, Cuba
Yassel Ramos: Proteomics Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana, 10600, Cuba
Vladimir Besada: Proteomics Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana, 10600, Cuba
Yeosvany Cabrera: Research and Development Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 83, Sancti Spíritus 60200, Cuba
Mario Pablo Estrada: Animal Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana 10600, Cuba; Corresponding authors.
Yamila Carpio: Animal Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana 10600, Cuba; Corresponding authors.

Journal volume & issue: Vol. 4
p. 100093

Abstract

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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.

Published in Fish and Shellfish Immunology Reports

ISSN: 2667-0119 (Online)
Publisher: Elsevier
Country of publisher: United Kingdom
LCC subjects: Science: Zoology
Website: https://www.journals.elsevier.com/fish-and-shellfish-immunology-reports/

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