Expansion of fish CCL20_like chemokines by genome and local gene duplication : Characterisation and expression analysis of 10 CCL20_like chemokines in rainbow trout (Oncorhynchus mykiss)

Acknowledgements Fuguo Liu was supported by a Newton International Fellowship funded by the Academy of Medical Sciences, UK (AMS, NIF004\1036). Tingyu Wang was supported by the Ministry of Science and Technology, Republic of China (Taiwan) (MOST 107-2917-I-564-019). YH was supported by a PhD Studentship from the Ministry of Education, Republic of China. Guangming Tian was supported financially by the State Scholarship Fund organised by the China Scholarship Council (201808420042).Peer reviewedPostprin

The expression of brown fat associated proteins in colorectal cancer and the relationship of uncoupling protein 1 with prognosis

© 2019 UICC Funding Innovate UK, NHS Grampian endowment funds and Vertebrate Antibodies Ltd. Acknowledgements The immunohistochemistry was performed with the support of the Grampian Biorepository (www.biorepository.nhsgrampian.org/). The antibodies were developed in collaboration with Vertebrate Antibodies Ltd (www.vertebrateantibodies.com) from whom they are now available commercially.Peer reviewedPostprin

Interleukin (IL)-2 Is a Key Regulator of T Helper 1 and T Helper 2 Cytokine Expression in Fish : Functional Characterization of Two Divergent IL2 Paralogs in Salmonids

This work was funded by the Biotechnology and Biological Sciences Research Council (BBSRC, BB/N024052/1) under the Newton Fund RCUK-CONICYT Research Partnerships call. YH was supported by a PhD Studentship from the Ministry of Education, Republic of China (Taiwan). EW was supported financially by the Faculty of Technology, Mahasarakham University Grant Year 2018. FL was supported by a Newton International Fellowship funded by the Academy of Medical Sciences, UK (AMS, NIF004\1036). ML and QX were supported financially by the National Scholarship Council of China. This work was partially supported financially by European Commission contract No. 311993 (TargetFish).Peer reviewedPublisher PD

Dysregulation of B Cell Activity During Proliferative Kidney Disease in Rainbow Trout

This work was supported by the European Research Council (ERC Consolidator Grant 2016 725061 TEMUBLYM) and the European Commission under the H2020 Programme (Grant H2020-634429 ParaFishControl). IE was recipient of APOSTD/2016/037 grant by the “Generalitat Valenciana” and YH was recipient of a PhD Studentship from the Ministry of Education, Republic of China (Taiwan). JWH was supported by BBSRC grant BB/K009125/1 and SNSF grant CRSII3_147649-1. PDR was funded by grant T1-BIO-1672 from the “Comunidad de Madrid”.Peer reviewedPublisher PD

Characterisation of rainbow trout peripheral blood leucocytes prepared by hypotonic lysis of erythrocytes, and analysis of their phagocytic activity, proliferation and response to PAMPs and proinflammatory cytokines

This work was funded by the Biotechnology and Biological Sciences Research Council (BBSRC, BB/N024052/1) under the Newton Fund RCUK-CONICYT Research Partnership. YH was supported by a PhD Studentship from the Ministry of Education, Republic of China (Taiwan). PAS was supported by the Newton-Bhabha PhD placement programme (268692473) and FL was supported by a Newton International Fellowship funded by the Academy of Medical Sciences (AMS, NIF004\1036). MI was funded by Fondecyt 1161015 and KM was funded by Fondecyt 11171057.Peer reviewedPostprin

Induction of IL-22 protein and IL-22-producing cells in rainbow trout Oncorhynchus mykiss

This work was funded by the Biotechnology and Biological Sciences Research Council (BBSRC, BB/N024052/1 and BB/R008442/1). This research was also funded by the European Commission under the 7th Framework Programme for Research and Technological Development (FP7) of the European Union (grant agreement No. 311993 TARGETFISH). YH was supported by a PhD Studentship from the Ministry of Education, Republic of China (Taiwan). Tingyu Wang was supported by the Ministry of Science and Technology, Republic of China (Taiwan) (MOST 107-2917-I-564-019). Fuguo Liu was supported by a Newton International Fellowship funded by the Academy of Medical Sciences, UK (AMS, NIF004\1036). Thanks also go to Dr. Dawn Shewring for excellent technical assistance and to Dr. Alex Douglas and Ms. Anna Harte for statistical advice.Peer reviewedPublisher PD

Characterisation and analysis of IFN-gamma producing cells in rainbow trout Oncorhynchus mykiss

11 Pág. Centro de Investigación en Sanidad Animal (CISA)IFN-γ is one of the key cytokines involved in Th1 immune responses. It is produced mainly by T cells and NK cells, which drive both innate and adaptive responses to promote protection against infections. IFN-γ orthologues have been discovered to be functionally conserved in fish, suggesting that type I immunity is present in early vertebrates. However, few studies have looked at IFN-γ protein expression in fish and its role in cell mediated immunity due to a lack of relevant tools. In this study, four monoclonal antibodies (mAbs) V27, N2, VAB3 and V91 raised against short salmonid IFN-γ peptides were developed and characterised to monitor IFN-γ expression. The results show that the IFN-γ mAbs specifically react to their peptide immunogens, recognise E. coli produced recombinant IFN-γ protein and rainbow trout IFN-γ produced in transfected HEK 293 cells. The mAb VAB3 was used further, to detect IFN-γ at the cellular level after in vitro and in vivo stimulation. In flow cytometry, a basal level of 3-5% IFN-γ secreting cells were detected in peripheral blood leucocytes (PBL), which increased significantly when stimulated in vitro with PAMPs (Aeromonas salmonicida bacterin), a mitogen (PHA) and recombinant cytokine (IL-2). Similarly, after injection of live bacteria (Aeromonas salmonicida) or poly I:C the number of IFN-γ+ cells increased in the lymphoid population of PBL, as well as in the myeloid population after infection, with the myeloid cells increasing substantially after both treatments. Immunohistochemistry was used to visualise the IFN-γ+ cells in spleen and head kidney following vaccination, which increased in intensity of staining and number relative to tissue from saline-injected control fish. These results show that several types of cells can produce IFN-γ in trout, and that they increase following infection or vaccination, and likely contribute to immune protection. Hence monitoring IFN-γ producing cells/protein secretion may be an important means to assess the effectiveness of Th1 responses and cell mediated immunity in fish.This research was funded by the European Commission under the 7th Framework Programme for Research and Technological Development (FP7) of the European Union (grant agreement No. 311993 TARGETFISH). YH was supported by a PhD Studentship from the Ministry of Education, Republic of China. TW was supported by the MASTS pooling initiative ( The Marine Alliance for Science and Technology for Scotland). MASTS is funded by the Scottish Funding Council (grant reference number HR09011) and contributing institutions.Peer reviewe

Dysregulation of B cell activity during proliferative kidney disease in rainbow trout

Funding This work was supported by the European Research Council (ERC Consolidator Grant 2016 725061 TEMUBLYM) and the European Commission under the H2020 Programme (Grant H2020-634429 ParaFishControl). IE was recipient of APOSTD/2016/037 grant by the “Generalitat Valenciana” and YH was recipient of a PhD Studentship from the Ministry of Education, Republic of China (Taiwan). JWH was supported by BBSRC grant BB/K009125/1 and SNSF grant CRSII3_147649-1. PDR was funded by grant T1-BIO-1672 from the “Comunidad de Madrid”.Proliferative kidney disease (PKD) is a widespread disease caused by the endoparasite Tetracapsuloides bryosalmonae (Myxozoa: Malacosporea). Clinical disease, provoked by the proliferation of extrasporogonic parasite stages, is characterized by a chronic kidney pathology with underlying transcriptional changes indicative of altered B cell responses and dysregulated T-helper cell-like activities. Despite the relevance of PKD to European and North American salmonid aquaculture, no studies, to date, have focused on further characterizing the B cell response during the course of this disease. Thus, in this work, we have studied the behavior of diverse B cell populations in rainbow trout (Oncorhynchus mykiss) naturally infected with T. bryosalmonae at different stages of preclinical and clinical disease. Our results show a clear upregulation of all trout immunoglobulins (Igs) (IgM, IgD, and IgT) demonstrated by immunohistochemistry and Western blot analysis, suggesting the alteration of diverse B cell populations that coexist in the infected kidney. Substantial changes in IgM, IgD, and IgT repertoires were also identified throughout the course of the disease further pointing to the involvement of the three Igs in PKD through what appear to be independently regulated mechanisms. Thus, our results provide strong evidence of the involvement of IgD in the humoral response to a specific pathogen for the first time in teleosts. Nevertheless, it was IgT, a fish-specific Ig isotype thought to be specialized in mucosal immunity, which seemed to play a prevailing role in the kidney response to T. bryosalmonae. We found that IgT was the main Ig coating extrasporogonic parasite stages, IgT+ B cells were the main B cell subset that proliferated in the kidney with increasing kidney pathology, and IgT was the Ig for which more significant changes in repertoire were detected. Hence, although our results demonstrate a profound dysregulation of different B cell subsets during PKD, they point to a major involvement of IgT in the immune response to the parasite. These results provide further insights into the pathology of PKD that may facilitate the future development of control strategiesDepto. de Genética, Fisiología y MicrobiologíaFac. de Ciencias BiológicasTRUEpu