Role of AKT1 & SIVA1 in resistance to avian salmonellosis
Lead Research Organisation:
The Pirbright Institute
Department Name: Avian Infectious Diseases
Abstract
Abstracts are not currently available in GtR for all funded research. This is normally because the abstract was not required at the time of proposal submission, but may be because it included sensitive information such as personal details.
Technical Summary
Avian resistance to systemic salmonellosis is largely mediated by the SAL1 locus. Analysis of the segregation of SNPs in the progeny of a sixth generation back-cross of inbred lines that differ in resistance (6 and 15I), as well as F(13) inter-cross lines, has fine-mapped the QTL to a 0.4Mb region encoding AKT1 and SIVA1. In murine and human cells, these proteins have opposing effects on apoptosis and induction of innate immunity. In mammalian cells, Salmonella activates AKT via injection of SopB. This delays apoptosis and promotes net intracellular replication in vitro and in mice, as evidenced by studies with sopB mutants, null mice and specific inhibitors. By contrast, SIVA1 is pro-apoptotic and is proposed to control Salmonella via release of heterophil extracellular traps upon cell death. No evidence exists that these mechanisms are relevant in chickens and a need exists to unravel how resistance-associated SNPs exert their effect.
We propose to define the kinetics of AKT1 and SIVA1 activation upon Salmonella infection using specific antisera, and associate this with the fate of infected cells ex vivo and in chickens by flow cytometry and immuno-staining. We will examine if levels or activities of AKT1 and SIVA1 explain the differential resistance of lines 6 and 15I. Net replication of Salmonella in such birds will be quantified in cells and tissues by fluorescence dilution, and visualised relative to AKT1 activation and apoptosis. We will further define if AKT1 plays a protective role in avian responses to Salmonella by use of sopB mutants, inhibitors and siRNA-mediated knock-down. Toward marker-assisted selection, we will survey the frequency of informative SNPs in a population of Hy-Line layers directly related to that exposed to a recent fowl typhoid outbreak. Birds of defined genotype will be challenged to associate genetic variation with AKT1/SIVA1 levels or activation, host responses and Salmonella replication.
We propose to define the kinetics of AKT1 and SIVA1 activation upon Salmonella infection using specific antisera, and associate this with the fate of infected cells ex vivo and in chickens by flow cytometry and immuno-staining. We will examine if levels or activities of AKT1 and SIVA1 explain the differential resistance of lines 6 and 15I. Net replication of Salmonella in such birds will be quantified in cells and tissues by fluorescence dilution, and visualised relative to AKT1 activation and apoptosis. We will further define if AKT1 plays a protective role in avian responses to Salmonella by use of sopB mutants, inhibitors and siRNA-mediated knock-down. Toward marker-assisted selection, we will survey the frequency of informative SNPs in a population of Hy-Line layers directly related to that exposed to a recent fowl typhoid outbreak. Birds of defined genotype will be challenged to associate genetic variation with AKT1/SIVA1 levels or activation, host responses and Salmonella replication.
Planned Impact
Please see document submitted by lead applicant (joint ref. M1529404)
People |
ORCID iD |
| Mark Fife (Principal Investigator) |
Publications
Batra A
(2017)
Selection of reference genes for gene expression analysis by real-time qPCR in avian cells infected with infectious bronchitis virus.
in Avian pathology : journal of the W.V.P.A
Crooijmans RP
(2013)
Large scale variation in DNA copy number in chicken breeds.
in BMC genomics
Gibson MS
(2012)
Identification, cloning, and functional characterization of the IL-1 receptor antagonist in the chicken reveal important differences between the chicken and mammals.
in Journal of immunology (Baltimore, Md. : 1950)
Gibson MS
(2012)
Identification, cloning and characterisation of interleukin-1F5 (IL-36RN) in the chicken.
in Developmental and comparative immunology
Kranis A
(2013)
Development of a high density 600K SNP genotyping array for chicken.
in BMC genomics
Psifidi A
(2016)
The genomic architecture of resistance to Campylobacter jejuni intestinal colonisation in chickens.
in BMC genomics
Psifidi A
(2018)
The Genomic Architecture of Fowl Typhoid Resistance in Commercial Layers.
in Frontiers in genetics
Staines K
(2016)
A Versatile Panel of Reference Gene Assays for the Measurement of Chicken mRNA by Quantitative PCR.
in PloS one
Te Pas MF
(2012)
Meta-analysis of chicken--salmonella infection experiments.
in BMC genomics
Thanh-Son T
(2012)
A maximum likelihood QTL analysis reveals common genome regions controlling resistance to Salmonella colonization and carrier-state.
in BMC genomics
| Description | This research is an ongoing collabroation with the Roslin Insititute and has identfied a gene involved in Salmonella disease resistance in chickens. We have identified the host gene in chickens that interacts with the bacterial protien SopB. Patent filed and continuing research with indutrial support. |
| Exploitation Route | Role of AKT1 and SIVA1 in resistance to avian salmonellosis is progressing well. We have identified the host gene in chickens that interacts with the bacterial protien SopB. Patent filed and continuing research with indutrial support. |
| Sectors | Agriculture, Food and Drink,Healthcare,Pharmaceuticals and Medical Biotechnology |
| URL | http://www.pirbright.ac.uk/research/GeneticsGenomics/Default.aspx |
| Description | This research is an ongoing collabroation with the Roslin Insititute and has identfied a gene involved in Salmonella disease resistance in chickens. "SAL1 Patent for Salmonella resistance in chickens"-. PCT/GB2010/000850. The Pirbright Institute. |
| Sector | Agriculture, Food and Drink,Healthcare,Pharmaceuticals and Medical Biotechnology |
| Impact Types | Economic |
| Description | Contributed to Chief Medical Officers report- Genomics section |
| Geographic Reach | Europe |
| Policy Influence Type | Implementation circular/rapid advice/letter to e.g. Ministry of Health |
| URL | https://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/6310... |
| Description | Commercial Partner |
| Amount | £30,000 (GBP) |
| Organisation | HyLine |
| Sector | Private |
| Country | United States |
| Start | 08/2014 |
| End | 06/2015 |
| Title | A Versatile Panel of Reference Gene Assays for the Measurement of Chicken mRNA by Quantitative PCR |
| Description | Quantitative real-time PCR assays are widely used for the quantification of mRNA within avian experimental samples. Multiple stably-expressed reference genes can be used to control random technical variation between samples. It is necessary to select reference genes with the lowest variation in representative samples. The candidate reference gene assays must be reliable. In particular, they should have high amplification specificity and efficiency, and not produce signals from contaminating DNA. Whilst recent research papers identify specific genes that are stable in particular tissues and experimental treatments, here we describe a panel of ten avian gene primer and probe sets that can be used to identify suitable reference genes in many experimental contexts. The panel was tested with TaqMan and SYBR Green systems in two experimental scenarios, a tissue collection, and virus infection of cultured fibroblasts. GeNorm and NormFinder algorithms were able to select appropriate reference gene sets in each case. We show the effects of using the selected genes on the detection of statistically significant differences in expression. The results are compared with those obtained using 28s ribosomal RNA, the present most widely accepted reference gene in chicken work, identifying circumstances where the use of this gene might provide misleading results. Widely used methods for eliminating DNA contamination of RNA reduced, but did not completely remove, detectable DNA. We therefore attached special importance to testing each qPCR assay for absence of signal using DNA template. |
| Type Of Material | Technology assay or reagent |
| Year Produced | 2016 |
| Provided To Others? | Yes |
| Impact | The assays and analyses developed here provide a useful resource for selecting reference genes for investigations of avian biology. |
| Title | SAL Locus |
| Description | SAL1 Patent for Salmonella resistance in chickens"-. PCT/GB2010/000850. The Pirbright Institute. |
| Type Of Material | Model of mechanisms or symptoms - non-mammalian in vivo |
| Year Produced | 2013 |
| Provided To Others? | Yes |
| Impact | SAL1 Patent for Salmonella resistance in chickens"-. PCT/GB2010/000850. The Pirbright Institute. |
| Title | Avian 600k SNP Array |
| Description | Development of a high density 600K SNP genotyping array for chicken. |
| Type Of Material | Data analysis technique |
| Year Produced | 2013 |
| Provided To Others? | Yes |
| Impact | Publication: Kranis A, Almas A Gheyas, Boschiero C, Turner F, Yu L, Smith S, Talbot R, Pirani A, Brew F, Kaiser P, Hocking P, Fife M et al. Development of a high density 600K SNP genotyping array for chicken. (2013) BMC Genomics, 14:59 |
| Description | Roslin |
| Organisation | University of Edinburgh |
| Department | The Roslin Institute |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Joint Grant: 'Role of AKT1 & SIVA1 in resistance to avian salmonellosis' Stevens, Kaiser & Fife |
| Collaborator Contribution | Poultry are a key reservoir of human Salmonella infections owing to the ability of some strains to colonise the avian intestines and reproductive tract. Birds often carry the bacteria in the absence of overt symptoms; however some types of Salmonella cause severe typhoid-like diseases in poultry that exert substantial welfare and economic costs. Global population growth and rising affluence are fuelling demand for poultry meat and eggs, and a need exists to enhance the supply and safety of such. Though vaccines are used in layers in some countries, most of the 55 billion chickens reared annually worldwide lack protection against Salmonella infection. We and others have discovered that some chickens exhibit heritable differences in resistance to Salmonella. It may be feasible to selectively breed for birds with improved resilience to Salmonella infection; however this requires the identification of resistance-associated factors and knowledge of how they act. By analysing the genetic material of birds that differ in resistance, we have located a region of the chicken chromosome that confers protection against typhoidal salmonellosis, both in laboratory studies and commercial poultry populations. Recent studies have now resolved the region associated with resistance to just a handful of genes. It is highly plausible that variation affecting two genes in this region (AKT1 and SIVA1) explains why birds react to Salmonella in different ways, as the encoded proteins control host processes that impact on the fate of bacteria. For example, AKT1 and SIVA1 control the death of infected cells and the induction of immune responses, but have opposing activities. It is not possible for us to predict how the genetic changes affecting these genes will alter their expression or activity. Moreover, it is unclear how such factors may control the growth and spread of Salmonella in birds. We therefore propose to: 1. Examine if birds normally respond to Salmonella infection by activating the expression or function of AKT1 and SIVA1. We will examine this in cells cultured from chickens, as well as in intact birds, and associate any differences with host responses and the fate of the bacteria. 2. Examine if lines of chicken known to differ in resistance to Salmonella infection vary in the levels or activation of AKT1 and SIVA1. 3. Use specific inhibitors and bacterial strains to establish that AKT1 activation is necessary for Salmonella to grow and spread in birds. 4. Define the nature, frequency and consequences of genetic changes affecting AKT1 and SIVA1 in commercial poultry populations. This will aid the selective breeding of chickens that show improved resilience to Salmonella infection. We are fortunate to have the support of one of the world's largest poultry breeding companies (Erich Wesjohann Group), who will provide birds, genome sequences, expertise and 10% of total project costs. This reflects the value of the proposed studies to the industry. The consortium has productively collaborated and the proposed studies are a timely, logical and feasible extension of our recent joint research. |
| Impact | SAL1 Patent for Salmonella resistance in chickens"-. PCT/GB2010/000850. The Pirbright Institute. |
| Start Year | 2012 |
| Title | RESISTANCE TO BACTERIAL INFECTION |
| Description | The present invention provides a method of identifying an animal having a genotype associated with resistance to bacterial infection comprising the steps of: (a) providing a sample from said animal; (b) determining the alleles at one or more markers of the SAL1 locus to identify the genotype of the marker, wherein said SAL1 locus lies between 54.0 MB to 54.8 MB of chicken Chromosome 5 or an equivalent thereof; and (c) determining whether the genotype is a genotype associated with resistance to bacterial infection. |
| IP Reference | WO2010125345 |
| Protection | Patent application published |
| Year Protection Granted | 2010 |
| Licensed | No |
| Impact | Secured further funding and allowed Link grant with commercial company. |
| Description | AB International Nidovirus Symposium |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | Batra, A., Maier, H.J., Britton P., Hiscox, J.A., Fife, M.S., 2014. AKT activation during infectious bronchitis virus infection. XIIIth International Nidovirus Symposium, Salamanca, Spain (Poster presentation) |
| Year(s) Of Engagement Activity | 2014 |
| Description | Chair of organising Committee for Wellcome Trust-Animal Genetics and Diseases 2017 |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | This meeting brought together specialists working on the interface between genomics, genetic engineering and infectious disease with the aims of improving animal and human health and welfare. Scientific sessions included: Genetics of immune responses and disease resistance Genetically engineered livestock (including genome editing) Quantitative genetics and epigenetics applied to disease Epidemiology and pathogen evolution Bioinformatics, comparative and functional genomics Precision medicine of animal companions |
| Year(s) Of Engagement Activity | 2017 |
| URL | https://coursesandconferences.wellcomegenomecampus.org/events/item.aspx?e=635&dm_i=2SUU,HOGH,4R4AW1,... |
| Description | Holt School Surrey. Bee meadow & A-level outreach Talk. |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Schools |
| Results and Impact | I discussed the importance of pollinators for biodiversity and food security with the younger students. I also discussed careers in science with A-Level students. There was good interaction and many questions surrounding both topics. |
| Year(s) Of Engagement Activity | 2018 |
| Description | Innovate Guildford Outreach (12th March) |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Public/other audiences |
| Results and Impact | With amazing exhibits, activities, workshops, talks and competitions, it's FREE to attend. Explore the four zones: Engineering, One Health, Digital Media and Future Living, experience interactive performances, hear inspiring speakers (including leading gaming pioneer Peter Molyneux and animal health pioneer Professor Nick Bacon) and take in an amazing atmosphere. Theatre, music and comedy will feature through the day, with everything from how technology might have affected Shakespeare's plays to risky recycling and photographic secrets. |
| Year(s) Of Engagement Activity | 2017 |
| URL | https://www.guildford.gov.uk/innovateguildford |
| Description | Kingdown School Warminster Wiltshire talk to 6th form students about career in science. |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Schools |
| Results and Impact | I presented my career path to the students to engage them on STEM careers. I had a good level of discussion and many questions. |
| Year(s) Of Engagement Activity | 2018 |
| Description | MF AIRG |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | Avina immunology research group meeting in Canada |
| Year(s) Of Engagement Activity | 2014 |
| Description | MF PAG |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | The Plant and Animal Genome XXIV Conference (PAG) is designed to provide a forum on recent developments and future plans for plant and animal genome projects. Consisting of technical presentations, poster sessions, exhibits and workshops, the conference is an excellent opportunity to exchange ideas and applications on this internationally important project. |
| Year(s) Of Engagement Activity | 2014,2016 |
| Description | Organiser of the Animal Genetics and Diseases conference 08 - 10 May 2019 Wellcome Genome Campus, UK. Highlighting recent advances in animal genetics and genomic technologies. |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | I am on the organising committee for this conference. The second conference in this series will highlight recent advances in animal genetics and genomic technologies. It will bring together specialists working on the interface of genomics, genetic engineering and infectious disease with the aims of improving animal and human health and welfare. Novel genomic technologies, mathematical modelling and quantitative genetics approaches, applied to host animals, as well as their pathogens, have transformed the understanding of animal diseases, host-pathogen interactions and epidemiology and their effects on productivity of farmed animal species and food supply chains. This year's conference will not only put the spotlight on the immune response of host animals and epidemiology but also cover the genetics and genomics of pathogens and the impact of animal-human relationships. We encourage registrations from researchers, breeders and technical specialists interested in learning and disseminating the latest cutting-edge techniques and methodologies across model species, wildlife, farmed animals and companion animals. |
| Year(s) Of Engagement Activity | 2019 |
| URL | https://coursesandconferences.wellcomegenomecampus.org/our-events/animal-genetics-diseases-2019/?dm_... |
| Description | Winston Churchill Careers Fair. Winston Churchill School, Hermitage Road, St Johns, Woking, Surrey, GU21 8TL |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Schools |
| Results and Impact | AS attended and presented at this event to stimulate increased interest in science and research. |
| Year(s) Of Engagement Activity | 2018 |