Plasma proteome responses in salmonid fish following immunization

Fiona K. Bakke; Milena M. Monte; David A. Stead; Dwight R. Causey; Alex Douglas; Daniel J. Macqueen; Helen Dooley

doi:10.3389/fimmu.2020.581070

Plasma proteome responses in salmonid fish following immunization

Fiona K. Bakke, Milena M. Monte, David A. Stead, Dwight R. Causey, Alex Douglas, Daniel J. Macqueen^*, Helen Dooley^* (Corresponding Author)

^*Corresponding author for this work

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Abstract

Vaccination plays a critical role in the protection of humans and other animals from infectious diseases. However, the same vaccine often confers different protection levels among individuals due to variation in genetics and/or immunological histories. While this represents a well-recognized issue in humans, it has received little attention in fish. Here we address this knowledge gap in a proteomic study of rainbow trout (Oncorhynchus mykiss, Walbaum), using non-lethal repeated blood sampling to establish the plasma protein response of individual fish following immunization. Six trout were immunized with adjuvanted hen egg-white lysozyme (HEL) and peripheral blood sampled at ten time points from day 0 to day 84 post-injection. We confirm that an antigen-specific antibody response to HEL was raised, showing differences in timing and magnitude among individuals. Using label-free liquid chromatography-mass spectrometry, we quantified the abundance of 278 plasma proteins across the timecourse. As part of the analysis, we show that this approach can distinguish many (but not all) duplicated plasma proteins encoded by paralogous genes retained from the salmonid-specific whole genome duplication event. Global variation in the plasma proteome was predominantly explained by individual differences among fish. However, sampling day explained a major component of variation in abundance for a statistically defined subset of 41 proteins, representing 15% of those detected. These proteins clustered into five groups showing distinct temporal responses to HEL immunization at the population level, and include classical immune (e.g. complement system members) and acute phase molecules (e.g. apolipoproteins, haptoglobins), several enzymes and other proteins supporting the immune response, in addition to evolutionarily conserved molecules that are as yet uncharacterized. Overall, this study improves our understanding of the fish plasma proteome, provides valuable marker proteins for different phases of the immune response, and has implications for vaccine development and the design of immune challenge experiments.

Original language	English
Article number	581070
Pages (from-to)	1-20
Number of pages	20
Journal	Frontiers in Immunology
Volume	11
DOIs	https://doi.org/10.3389/fimmu.2020.581070
Publication status	Published - 8 Oct 2020

Bibliographical note

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.

Keywords

immunity
immunoglobulin M
liquid chromatography-mass spectrometry
plasma
proteome
salmonid
trout

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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Cite this

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title = "Plasma proteome responses in salmonid fish following immunization",

abstract = "Vaccination plays a critical role in the protection of humans and other animals from infectious diseases. However, the same vaccine often confers different protection levels among individuals due to variation in genetics and/or immunological histories. While this represents a well-recognized issue in humans, it has received little attention in fish. Here we address this knowledge gap in a proteomic study of rainbow trout (Oncorhynchus mykiss, Walbaum), using non-lethal repeated blood sampling to establish the plasma protein response of individual fish following immunization. Six trout were immunized with adjuvanted hen egg-white lysozyme (HEL) and peripheral blood sampled at ten time points from day 0 to day 84 post-injection. We confirm that an antigen-specific antibody response to HEL was raised, showing differences in timing and magnitude among individuals. Using label-free liquid chromatography-mass spectrometry, we quantified the abundance of 278 plasma proteins across the timecourse. As part of the analysis, we show that this approach can distinguish many (but not all) duplicated plasma proteins encoded by paralogous genes retained from the salmonid-specific whole genome duplication event. Global variation in the plasma proteome was predominantly explained by individual differences among fish. However, sampling day explained a major component of variation in abundance for a statistically defined subset of 41 proteins, representing 15% of those detected. These proteins clustered into five groups showing distinct temporal responses to HEL immunization at the population level, and include classical immune (e.g. complement system members) and acute phase molecules (e.g. apolipoproteins, haptoglobins), several enzymes and other proteins supporting the immune response, in addition to evolutionarily conserved molecules that are as yet uncharacterized. Overall, this study improves our understanding of the fish plasma proteome, provides valuable marker proteins for different phases of the immune response, and has implications for vaccine development and the design of immune challenge experiments.",

keywords = "immunity, immunoglobulin M, liquid chromatography-mass spectrometry, plasma, proteome, salmonid, trout",

author = "Bakke, {Fiona K.} and Monte, {Milena M.} and Stead, {David A.} and Causey, {Dwight R.} and Alex Douglas and Macqueen, {Daniel J.} and Helen Dooley",

note = "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{\textquoteright}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.",

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AU - Stead, David A.

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AU - Macqueen, Daniel J.

AU - Dooley, Helen

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Plasma proteome responses in salmonid fish following immunization

Abstract

Bibliographical note

Keywords

UN SDGs

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David Stead

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Plasma proteome responses in salmonid fish following immunization

Abstract

Bibliographical note

Keywords

UN SDGs

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David Stead

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