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      RNA sequencing-based analysis of the spleen transcriptome following infectious bronchitis virus infection of chickens selected for different mannose-binding lectin serum concentrations

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          Abstract

          Background

          Avian infectious bronchitis is a highly contagious disease of the upper-respiratory tract caused by infectious bronchitis virus (IBV). Understanding the molecular mechanisms involved in the interaction between innate and adaptive immune responses to IBV infection is a crucial element for further improvements in strategies to control IB. To this end, two chicken lines, selected for high (L10H line) and low (L10L line) serum concentration of mannose-binding lectin (MBL) were studied. In total, 32 birds from each line were used. Sixteen birds from each line were infected with IBV and sixteen were left uninfected. Eight uninfected and infected birds from each line were euthanized at 1 and 3 weeks post infection. RNA sequencing was performed on spleen samples from all 64 birds and differential gene expression analysis was performed for four comparisons: L10L line versus L10H line for uninfected birds at weeks 1 and 3, respectively, and in the same way for infected birds. Functional analysis was performed using Gene Ontology (GO) Immune System Process terms specific for Gallus gallus.

          Results

          Comparing uninfected L10H and L10L birds, we identified 1698 and 1424 differentially expressed (DE) genes at weeks 1 and 3, respectively. For the IBV-infected birds, 1934 and 866 DE genes were identified between the two lines at weeks 1 and 3, respectively. The two most enriched GO terms emerging from the comparison of uninfected birds between the two lines were “Lymphocyte activation involved in immune response” and “Somatic recombination of immunoglobulin genes involved in immune response” at weeks 1 and 3, respectively. When comparing IBV-infected birds between the two lines, the most enriched GO terms were “Alpha-beta T cell activation” and “Positive regulation of leukocyte activation” at weeks 1 and 3, respectively.

          Conclusions

          Healthy birds from the two lines showed significant differences in expression profiles for subsets of adaptive and innate immunity-related genes, whereas comparison of the IBV-infected birds from the two lines showed differences in expression of immunity-related genes involved in T cell activation and proliferation. The observed transcriptome differences between the two lines indicate that selection for MBL had influenced innate as well as adaptive immunity.

          Electronic supplementary material

          The online version of this article (doi:10.1186/s12864-016-2403-1) contains supplementary material, which is available to authorized users.

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          Controlling the False Discovery Rate: A Practical and Powerful Approach to Multiple Testing

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            Integration of biological networks and gene expression data using Cytoscape.

            Cytoscape is a free software package for visualizing, modeling and analyzing molecular and genetic interaction networks. This protocol explains how to use Cytoscape to analyze the results of mRNA expression profiling, and other functional genomics and proteomics experiments, in the context of an interaction network obtained for genes of interest. Five major steps are described: (i) obtaining a gene or protein network, (ii) displaying the network using layout algorithms, (iii) integrating with gene expression and other functional attributes, (iv) identifying putative complexes and functional modules and (v) identifying enriched Gene Ontology annotations in the network. These steps provide a broad sample of the types of analyses performed by Cytoscape.
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              FastQC: a quality-control tool for high-throughput sequence data.

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                Author and article information

                Contributors
                edin.hamzic@jouy.inra.fr
                rikke.kjaerup@anis.au.dk
                nuria.mach@jouy.inra.fr
                giulietta.minozzi@unimi.it
                francesco.strozzi@ptp.it
                valentina.gualdi@ptp.it
                john.williams01@adelaide.edu.au
                jun.chen@cobb-vantress.com
                eva.wattrang@sva.se
                bart.buitenhuis@mbg.au.dk
                helle.juulmadsen@anis.au.dk
                tina.dalgaard@anis.au.dk
                Journal
                BMC Genomics
                BMC Genomics
                BMC Genomics
                BioMed Central (London )
                1471-2164
                27 January 2016
                27 January 2016
                2016
                : 17
                : 82
                Affiliations
                [ ]UMR1313 Animal Genetics and Integrative Biology Unit, AgroParisTech, Université Paris-Saclay, 16 rue Claude Bernard, 75005 Paris, France
                [ ]UMR1313 Animal Genetics and Integrative Biology Unit, INRA, Université Paris-Saclay, Domaine de Vilvert, 78350 Jouy-en-Josas, France
                [ ]Department of Molecular Biology and Genetics, Center for Quantitative Genetics and Genomics, Aarhus University, Blichers Allé 20, P.O. Box 50, 8830 Tjele, Denmark
                [ ]Department of Animal Science, Aarhus University, Blichers Allé 20, P.O. Box 50, 8830 Tjele, Denmark
                [ ]Parco Tecnologico Padano, Via Einstein, 26900 Lodi, Italy
                [ ]University of Milan, DIVET, Via Celoria 10, 20133 Milan, Italy
                [ ]School of Animal and Veterinary Sciences, University of Adelaide, SA 5371 Roseworthy, Australia
                [ ]Cobb-Vantress Inc, US-412 Road, Siloam Springs, AR 72761 USA
                [ ]National Veterinary Institute, Ulls väg 2B, 751 89 Uppsala, Sweden
                Article
                2403
                10.1186/s12864-016-2403-1
                4729133
                d8845fc1-1601-4009-953c-92149d671fe3
                © Hamzić et al. 2016

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 24 August 2015
                : 18 January 2016
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/501100000785, Education, Audiovisual and Culture Executive Agency (BE);
                Award ID: European Graduate School in Animal Breeding and Genetics
                Funded by: Cobb-Vantress, Inc.
                Funded by: FundRef http://dx.doi.org/10.13039/100007398, Strategiske Forskningsråd (DK);
                Award ID: 10-093534
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/100007398, Strategiske Forskningsråd (DK);
                Award ID: 10-093534
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/100007398, Strategiske Forskningsråd (DK);
                Award ID: 10-093534
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/501100004963, Seventh Framework Programme (BE);
                Award ID: FP7-228394
                Award Recipient :
                Categories
                Research Article
                Custom metadata
                © The Author(s) 2016

                Genetics
                ibv,coronavirus,infectious bronchitis,chicken,rna sequencing,transcriptome,spleen,mannose-binding lectin,immune response

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