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17. Search for effective oral adjuvants for rainbow trout (Oncorhynchus mykiss)

Author

Docando, Felix, Arense, Paula, Martín-Martín, Alba, Wang, Tiehui, Tafalla, Carolina, Díaz-Rosales, Patricia, Ministerio de Ciencia e Innovación (España), Comunidad de Madrid, Docando, Felix, Martín-Martín, Alba, Wang, Tiehui, Tafalla, Carolina, and Díaz-Rosales, Patricia

Subjects
Lipopolysaccharides, Mammals, LPS, beta-Glucans, General Medicine, Aquatic Science, CpGs, Fish Diseases, Rainbow trout, Adjuvants, Immunologic, Immunoglobulin M, Oncorhynchus mykiss, Animals, Environmental Chemistry, Adjuvants, Glucans, Flagellin
Abstract

6 Pág. Centro de Investigación en Sanidad Animal (CISA), Disease prevention by vaccination is, on economic, environmental and ethical grounds the most appropriate method for pathogen control currently available to the aquaculture sector. However, vaccine administration in aquatic animals faces obvious technical problems not encountered in other land animals. Thus, oral vaccines are highly demanded by the aquaculture sector that requests alternatives to the labor-intensive injectable vaccines that require individual handling of fish, provoking stress-related immunosuppression and handling mortalities. Despite this, most previous attempts to obtain effective oral vaccines have failed both in fish and mammals. This could be a consequence of very restricted tolerance mechanisms in the intestine given the fact that this mucosa is at the frontline upon antigen encounter and has to balance the delicate equilibrium between tolerance and immunity in a microbe rich aquatic environment. In this context, the search for an optimal combination of antigen and adjuvant that can trigger an adequate immune response able to circumvent intestinal tolerance is needed for each pathogen. To this aim, we have explored potential of molecules such as β-glucans, flagellin, CpG and bacterial lipopolysacharide (LPS) as oral adjuvants. For this, we have determined the effects of these adjuvants ex vivo in rainbow trout intestine tissue sections, and in vitro in leucocytes isolated from rainbow trout spleen and intestine. The effects were evaluated by analyzing the levels of transcription of different genes related to the innate and adaptive immune response, as well as evaluating the number of IgM-secreting cells. LPS seems to be the molecule with stronger immunostimulatory potential, and could safely be used as a mucosal adjuvant in rainbow trout. Moreover, the designed strategy provides a fast methodology to screen adjuvants that are suitable for oral vaccination, providing us with valuable information about how the intestinal mucosa is regulated in fish., This work was financed by the Spanish Ministry of Science, 2016 725061 TEMUBLYM Innovation and Universities (projects AGL2017-85494-C2-1-R and PID2020-113268RB-I00), by the Comunidad de Madrid (grant 2016-T1/BIO-1672 ) and the European Research Council (ERC Consolidator Grant ).

Published
2022

20. Molecular cloning, expression, structural and functional analysis of several immune relevant genes in rainbow trout Oncorhynchus mykiss

Author

Wang, Tiehui

Subjects
597, Rainbow trout diseases
Abstract

Fish diseases endanger the aquaculture industry worldwide. Approaches to combat fish diseases are hampered by the relative poor knowledge of immune relevant genes in fish. This thesis describes the cloning and expression of several immune relevant genes in rainbow trout (Oncorhynchus mykiss). The full-length cDNA and genomic DNA of the inducible nitric oxide synthase (iNOS) gene have been cloned and sequenced. The gene structure was the first to be determined outwith the mammals. The expression of the iNOS gene is induced by virus (VHSV) infection, but its detection by RT-PCR might be impaired by the secondary structure formed in the iNOS mRNA. Screening of a genomic library resulted in the isolation of a second allele of IL-1b1 gene with a large intron III. This allele appears to have resulted from a recent retroposition of a HpaI SINE into intron III. Two other retroposition events have also been recognised in the same intron. The survival mechanism of this SINE is discussed and a model of trout IL-1b gene formation proposed. Both alleles of the IL-1b1 gene are constitutively expressed and induced by LPS and recombinant trout IL-1b1 in heterozygous RTS-11 cells. The promoter of the IL-1b1 allele S gene was isolated and the transcription start site was determined. A series of promoter-reportor gene constructs were transfected into RTG cells to study their activities under different conditions. The first intron is an important part of the promoter of IL-1b1 allele S and NFkB is a transcription factor required for IL-1b1 expression. The cytokine receptor common gamma chain (gC) was also isolated from rainbow trout, the first outwith mammals. Its expression can be detected in blood, gill, kidney, brain and liver, and in macrophage cultures. The expression of gC in macrophage cell cultures is upregulated by recombinant IL-1b1 and LPS stimulation. The expression of gC is also detectable in RTG-2 cells with an increased transcript level after stimulation with recombinant IL-1b1.

Published
2002

23. Infectious pancreatic necrosis virus (IPNV) recombinant viral protein 1 (VP1) and VP2-Flagellin fusion protein elicit distinct expression profiles of cytokines involved in type 1, type 2, and regulatory T cell response in rainbow trout (Oncorhynchus mykiss)

Author

Wong-Benito, Valentina, primary, Barraza, Felipe, additional, Trujillo-Imarai, Agustín, additional, Ruiz-Higgs, Daniela, additional, Montero, Ruth, additional, Sandino, Ana María, additional, Wang, Tiehui, additional, Maisey, Kevin, additional, Secombes, Christopher J., additional, and Imarai, Mónica, additional

Published
2022
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25. Corrigendum: Type I Interferon Regulates the Survival and Functionality of B Cells in Rainbow Trout

Author

Benedicenti, Ottavia [0000-0003-4067-7617], Wang, Tiehui [0000-0002-1148-8959], Morel, Esther [0000-0002-4768-082X], Secombes, Christopher J. [0000-0001-8781-176X], Soleto, Irene [0000-0002-8005-2411], Díaz-Rosales, Patricia [0000-0001-6965-3702], Tafalla, Carolina [0000-0002-0860-2976], Benedicenti, Ottavia, Wang, Tiehui, Morel, Esther, Secombes, Christopher J., Soleto, Irene, Díaz-Rosales, Patricia, Tafalla, Carolina, Benedicenti, Ottavia [0000-0003-4067-7617], Wang, Tiehui [0000-0002-1148-8959], Morel, Esther [0000-0002-4768-082X], Secombes, Christopher J. [0000-0001-8781-176X], Soleto, Irene [0000-0002-8005-2411], Díaz-Rosales, Patricia [0000-0001-6965-3702], Tafalla, Carolina [0000-0002-0860-2976], Benedicenti, Ottavia, Wang, Tiehui, Morel, Esther, Secombes, Christopher J., Soleto, Irene, Díaz-Rosales, Patricia, and Tafalla, Carolina

Abstract

In the original article, there was a mistake in Figure 1 as published. Although the numbers were correct, all dot plots in Figure 1A were the same. The corrected Figure 1 appears below. The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated. © 2021 Benedicenti, Wang, Morel, Secombes, Soleto, Dı́az-Rosales and Tafalla. This corrects the article DOI: 10.3389/fimmu.2020.01494.

Published
2021

28. Distinct modes of action of CD40L and adaptive cytokines IL-2, IL-4/13, IL-10 and IL-21 on rainbow trout IgM+ B cells

Author

European Commission, Ministerio de Ciencia, Innovación y Universidades (España), Agencia Estatal de Investigación (España), Abós, Beatriz (0000-0002-3402-7397), Wang, Tiehui [0000-0002-1148-8959], Secombes, Christopher J. [0000-0001-8781-176X], Tafalla, Carolina [0000-0002-0860-2976], Abós, Beatriz, Wang, Tiehui, Secombes, Christopher J., Tafalla, Carolina, European Commission, Ministerio de Ciencia, Innovación y Universidades (España), Agencia Estatal de Investigación (España), Abós, Beatriz (0000-0002-3402-7397), Wang, Tiehui [0000-0002-1148-8959], Secombes, Christopher J. [0000-0001-8781-176X], Tafalla, Carolina [0000-0002-0860-2976], Abós, Beatriz, Wang, Tiehui, Secombes, Christopher J., and Tafalla, Carolina

Abstract

In mammals, conventional B (B2) cells are activated within lymphoid follicles through a close relationship with T follicular helper (Tfh) cells. The interaction between CD40 expressed on B cells and its ligand (CD40L) expressed on Tfh cells is a key signal that regulates the formation of germinal centers (GCs), B cell survival, proliferation and differentiation to plasma cells (PCs) or memory cells. Additionally, certain soluble cytokines produced by T cells also strongly condition the outcome of this interaction. Despite the many differences found between fish B cells and mammalian B2 cells, and the lack of conventional GCs, rainbow trout IgM+ B cells have been shown to be stimulated by CD40L, however, whether cytokines commonly produced by T cells can further modulate this response has never been addressed to date. Thus, in this study, we determined the effects of recombinant rainbow trout adaptive cytokines interleukin 2B (IL-2B), IL-4/13A, IL-4/13B, IL-10 and IL-21 (cytokines known to activate B cells in mammals) on splenic IgM+ B cells alone or in combination with CD40L. We studied how these cytokines and CD40L cooperated to promote IgM+ B cell survival, proliferation and IgM secretion. The results obtained provide valuable information for the first time in teleost fish on how different T cell signals cooperate to activate B cells in the absence of GCs.

Published
2020

34. Contributors

Author

Accorsi, Alice, primary, Armitage, Sophie A.O., additional, Ballarin, Loriano, additional, Barela Hudgell, Megan A., additional, Bird, Steve, additional, Brites, Daniela, additional, Buonocore, Francesco, additional, Castro, Rosario, additional, Cima, Francesca, additional, Doolittle, Russell F., additional, Franchi, Nicola, additional, Golconda, Preethi, additional, Kasahara, Masanori, additional, Malagoli, Davide, additional, Man Lun, Cheng, additional, Nakao, Miki, additional, Nonaka, Masaru, additional, Nonaka, Mayumi I., additional, Oren, Matan, additional, Ottaviani, Enzo, additional, Scapigliati, Giuseppe, additional, Secombes, Christopher J., additional, Smith, L. Courtney, additional, Smith, Valerie J., additional, Somamoto, Tomonori, additional, Sutoh, Yoichi, additional, Tafalla, Carolina, additional, van Putten, Jos P.M., additional, Vasta, Gerardo R., additional, Voogdt, Carlos G.P., additional, Wang, Tiehui, additional, and Zou, Jun, additional

Published
2016
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