Individual monitoring of immune responses in rainbow trout after cohabitation and intraperitoneal injection challenge with Yersinia ruckeri

Acknowledgements This work was funded by the National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs, grant G1100675). The authors are grateful to the aquarium staff at the University of Aberdeen (Karen Massie) and Dr David Smail at Marine Scotland for valuable discussion during the establishment of the experimental design.Peer reviewedPostprin

Plasma proteome responses in salmonid fish following immunization

Data Availability Statement The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found in the article/Supplementary Material. Ethics Statement The animal study was reviewed and approved by UK home office and University of Aberdeen’s Animal Welfare and Ethical Review Body (AWERB). Author Contributions Study conception and design: DM and HD. Animal work: MM and HD. Proteomics lab work: DS. Proteomic data analysis: FB, DC, AD. Data interpretation: FB, DM, and HD. Drafted figures and tables: FB and DM. Drafted manuscript: FB, DM, and HD. All authors contributed to the article and approved the submitted version. Funding This work was supported by the Biotechnology and Biological Sciences Research Council (BBSRC) grant numbers: BB/M010996/1, BB/M026345/1, BBS/E/D/20002174, and BBS/E/D/10002071. Conflict of Interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. Acknowledgments Our thanks to Prof. Chris Secombes (University of Aberdeen) for the 4C10 anti-salmonid IgM mAb used in our ELISAs and for his valuable intellectual contributions during the planning of this project. We also gratefully acknowledge the supervisory support given by Prof. Sam Martin (University of Aberdeen) to FB.Peer reviewedPublisher PD

Causes of blindness and vision impairment in 2020 and trends over 30 years, and prevalence of avoidable blindness in relation to VISION 2020: the Right to Sight: an analysis for the Global Burden of Disease Study

Background: Many causes of vision impairment can be prevented or treated. With an ageing global population, the demands for eye health services are increasing. We estimated the prevalence and relative contribution of avoidable causes of blindness and vision impairment globally from 1990 to 2020. We aimed to compare the results with the World Health Assembly Global Action Plan (WHA GAP) target of a 25% global reduction from 2010 to 2019 in avoidable vision impairment, defined as cataract and undercorrected refractive error. Methods: We did a systematic review and meta-analysis of population-based surveys of eye disease from January, 1980, to October, 2018. We fitted hierarchical models to estimate prevalence (with 95% uncertainty intervals [UIs]) of moderate and severe vision impairment (MSVI; presenting visual acuity from <6/18 to 3/60) and blindness (<3/60 or less than 10° visual field around central fixation) by cause, age, region, and year. Because of data sparsity at younger ages, our analysis focused on adults aged 50 years and older. Findings: Global crude prevalence of avoidable vision impairment and blindness in adults aged 50 years and older did not change between 2010 and 2019 (percentage change −0·2% [95% UI −1·5 to 1·0]; 2019 prevalence 9·58 cases per 1000 people [95% IU 8·51 to 10·8], 2010 prevalence 96·0 cases per 1000 people [86·0 to 107·0]). Age-standardised prevalence of avoidable blindness decreased by −15·4% [–16·8 to −14·3], while avoidable MSVI showed no change (0·5% [–0·8 to 1·6]). However, the number of cases increased for both avoidable blindness (10·8% [8·9 to 12·4]) and MSVI (31·5% [30·0 to 33·1]). The leading global causes of blindness in those aged 50 years and older in 2020 were cataract (15·2 million cases [9% IU 12·7–18·0]), followed by glaucoma (3·6 million cases [2·8–4·4]), undercorrected refractive error (2·3 million cases [1·8–2·8]), age-related macular degeneration (1·8 million cases [1·3–2·4]), and diabetic retinopathy (0·86 million cases [0·59–1·23]). Leading causes of MSVI were undercorrected refractive error (86·1 million cases [74·2–101·0]) and cataract (78·8 million cases [67·2–91·4]). Interpretation: Results suggest eye care services contributed to the observed reduction of age-standardised rates of avoidable blindness but not of MSVI, and that the target in an ageing global population was not reached. Funding: Brien Holden Vision Institute, Fondation Théa, The Fred Hollows Foundation, Bill & Melinda Gates Foundation, Lions Clubs International Foundation, Sightsavers International, and University of Heidelberg

Identification of IL-34 in teleost fish: differential expression of rainbow trout IL-34, MCSF1 and MCSF2, ligands of the MCSF receptor

11 páginas, 8 figuras, 2 tablasThe mononuclear phagocyte system is composed of monocytes, macrophages and dendritic cells and has crucial roles in inflammation, autoimmunity, infection, cancer, organ transplantation and in maintaining organismal homeostasis. Interleukin-34 (IL-34) and macrophage colony stimulating factor (MCSF), both signalling through the MCSF receptor, regulate the mononuclear phagocyte system. A single IL-34 and MCSF gene are present in tetrapods. Two types of MCSF exist in teleost fish which is resulted from teleost-wide whole genome duplication. In this report, we first identified and sequence analysed six IL-34 genes in five teleost fish, rainbow trout, fugu, Atlantic salmon, catfish and zebrafish. The fish IL-34 molecules had a higher identity within fish group but low identities to IL-34s from birds (27.2–33.8%) and mammals (22.2–31.4%). However, they grouped with tetrapod IL-34 molecules in phylogenetic tree analysis, had a similar 7 exon/6 intron gene organisation, and genes in the IL-34 loci were syntenically conserved. In addition, the regions of the four main helices, along with a critical N-glycosylation site were well conserved. Taken together these data suggest that the teleost IL-34 genes described in this report are orthologues of tetrapod IL-34. Comparative expression study of the three trout MCSFR ligands revealed that IL-34, MCSF1 and MCSF2 are differentially expressed in tissues and cell lines. The expression of MCSF1 and MCSF2 showed great variance in different tissues and cell lines, suggesting a role in the differentiation and maintenance of specific macrophage lineages in specific locations. The relatively high levels of IL-34 expression across different tissues suggests a homeostatic role of IL-34 for the macrophage lineage in fish. One striking observation in the present study was the lack of induction of MCSF1 and MCSF2 expression but the quick induction of IL-34 expression by PAMPs and inflammatory cytokines in cell lines and primary head kidney macrophages in rainbow trout. In a parasitic proliferative kidney disease (PKD) model, the expression of IL-34 but not the dominant MCSF2 was affected by PKD, suggesting an involvement of macrophage function in this disease model. Thus IL-34 expression is sensitive to inflammatory stimuli and may regulate macrophage biology once up-regulated.TW received funding from the MASTS pooling initiative (The Marine Alliance for Science and Technology for Scotland). TK was supported financially by the Program “Improvement of Research Environment for Young Researchers” from the Japanese Ministry of Education, Culture, Sports, Science and Technology, a Grant-in-Aid for Young Scientists (23780199) and a grant for Scientific Research on Priority Areas from the University of Miyazaki. MMC thanks the Consejo Superior de Investigaciones Científicas (CSIC, Spain) and the Xunta de Galicia for her “Ángeles Alvariño” postdoctoral contract. Thanks to Dr. Jason Holland (Scottish Fish Immunology Research Centre, University of Aberdeen) for supplying of the PKD samples.Peer reviewe

Identification of two FoxP3 genes in rainbow trout (Oncorhynchus mykiss) with differential induction patterns.

FoxP3 is a master transcription factor for the development and function of regulatory T cells in mammals, but little is known about this molecule in fish. Two paralogues of mammalian FoxP3 that share 83.9% identity at the amino acid level have been identified in rainbow trout (Oncorhynchus mykiss). The C-terminal region containing a Zn_C2H2 domain, a leucine zipper-like domain and a forkhead (FH) domain important for dimerization, nuclear translocation, and DNA binding, is well conserved between fish and other vertebrate FoxP3. However, the N-terminal of FoxP3 that is required for FoxP3-mediated repression of transcription is greatly diverged between fish, amphibians and monotreme mammals compared to eutherian mammals, suggesting that FoxP3 in fish, frog and platypus may have a different role to the human and mouse counterpart that defines the Treg cellular lineage and mediates the immune regulatory function. The expression of both trout (t) FoxP3a and tFoxP3b are detectable in all the 14 tissues examined without any significant difference except in muscle in which the expression of tFoxP3a was higher. Both tFoxP3a and tFoxP3b are highly expressed in thymus and in immune related organs including the spleen, kidney, gills and intestine, and are up-regulated by phytohaemagglutinin (PHA) in splenocytes and thymocytes. Whilst the up-regulated tFoxP3b expression induced by PHA was dose-dependent it required a higher PHA concentration to achieve maximal expression relative to tFoxP3a where the highest expression level was seen using 1 mug/ml PHA with higher concentrations having no further effects. In addition, the tFoxP3b expression increased during development from eyed eggs to fry, when it reached a comparable level to that of tFoxP3a. In contrast, tFoxP3a expression was at a high and almost constant level over all of the developmental stages examined. The high level of tFoxP3a expression in early development may be related to the relatively high constitutive level of tFoxP3a expression seen in muscle, perhaps suggesting novel roles of tFoxP3 in fish muscle. The structural and expression analysis suggests that the tFoxP3a and tFoxP3b are subject to differential modulation of expression and may have evolved novel functions. The identification of the two trout FoxP3 paralogues will help to clarify the existence of Treg cells and to dissect the T cell differentiation pathways in fish

Identification and expression modulation of a C-type lectin domain family 4 homologue that is highly expressed in monocytes/macrophages in rainbow trout (Oncorhynchus mykiss)

Acknowledgements PJ was supported by a PhD studentship from the Marine Alliance for Science and Technology for Scotland (MASTS), and TW was supported by a MASTS postdoctoral fellowship.Peer reviewedPostprintPostprin

Identification of IL-34 in teleost fish: differential expression of rainbow trout IL-34, MCSF1 and MCSF2, ligands of the MCSF receptor.

The mononuclear phagocyte system is composed of monocytes, macrophages and dendritic cells and has crucial roles in inflammation, autoimmunity, infection, cancer, organ transplantation and in maintaining organismal homeostasis. Interleukin-34 (IL-34) and macrophage colony stimulating factor (MCSF), both signalling through the MCSF receptor, regulate the mononuclear phagocyte system. A single IL-34 and MCSF gene are present in tetrapods. Two types of MCSF exist in teleost fish which is resulted from teleost-wide whole genome duplication. In this report, we first identified and sequence analysed six IL-34 genes in five teleost fish, rainbow trout, fugu, Atlantic salmon, catfish and zebrafish. The fish IL-34 molecules had a higher identity within fish group but low identities to IL-34s from birds (27.2-33.8%) and mammals (22.2-31.4%). However, they grouped with tetrapod IL-34 molecules in phylogenetic tree analysis, had a similar 7 exon/6 intron gene organisation, and genes in the IL-34 loci were syntenically conserved. In addition, the regions of the four main helices, along with a critical N-glycosylation site were well conserved. Taken together these data suggest that the teleost IL-34 genes described in this report are orthologues of tetrapod IL-34. Comparative expression study of the three trout MCSFR ligands revealed that IL-34, MCSF1 and MCSF2 are differentially expressed in tissues and cell lines. The expression of MCSF1 and MCSF2 showed great variance in different tissues and cell lines, suggesting a role in the differentiation and maintenance of specific macrophage lineages in specific locations. The relatively high levels of IL-34 expression across different tissues suggests a homeostatic role of IL-34 for the macrophage lineage in fish. One striking observation in the present study was the lack of induction of MCSF1 and MCSF2 expression but the quick induction of IL-34 expression by PAMPs and inflammatory cytokines in cell lines and primary head kidney macrophages in rainbow trout. In a parasitic proliferative kidney disease (PKD) model, the expression of IL-34 but not the dominant MCSF2 was affected by PKD, suggesting an involvement of macrophage function in this disease model. Thus IL-34 expression is sensitive to inflammatory stimuli and may regulate macrophage biology once up-regulated