Characterisation of the Pattern Recognition Receptors required for the development of protective immunity against Salmonella infection

Lead Research Organisation: University of Cambridge
Department Name: Veterinary Medicine

Abstract

Salmonella enterica causes a wide range of diseases in many animals. Economic losses to the farming industry through Salmonella infection are potentially very high, but also important is the fact that a number of serovars that infect animals can also cause food poisoning and gastroenteritis in humans. In mice, infection with Salmonella enterica serovar Typhimurium (S. Typhimurium) causes clinical signs that are very similar to those seen in invasive infections of humans and chickens. Infection studies in mice will therefore allow us to understand how infection is likely to develop and spread within other species of animals. Vaccination of animals to lower the levels of Salmonella in meat and eggs will reduce the chances of infection spreading to people. The mouse model of infection is also excellent for studying vaccines. A successful vaccine must be generate a strong immune response but must also be safe for application in young and older individuals. Understanding how Salmonella interacts with its host to generate an immune response is critical when trying to design new vaccines. The animal detects the presence of these bacterial molecules through specialised proteins called Pattern Recognition Receptors (PRRs) which then initiate host defence mechanisms to clear the infection. These receptors are also believed to be important in generating a good response to vaccines, but precisely how this happens is currently unclear. The knowledge on whether chickens, for example, use PRRs to control Salmonella infection is too poorly understood to perform these studies in this species, but we can use mice without PRRs to determine whether these proteins are important for the control of, and protection against, Salmonella. Here we will determine which PRRs are important in protecting mice against Salmonella infections. The impact of this work will be to determine which PRRs are important in generating successful protective immune responses against Salmonella and these data will therefore allow us to design better vaccines for chickens, other domestic animals and humans.

Technical Summary

Recognition of invading microorganisms by the host is an essential first step in the generation of protective immunity. Various microbial components are known to stimulate a group of receptors collectively known as the Pattern Recognition Receptors (PRR) that include the Toll-like Receptors (TLRs) and Nucleotide-binding domain, Leucine-rich repeat containing Receptors (NLRs). TLRs and NLRs respond to microbial components in different cellular sites and co-ordinated engagement of these receptors leads to intense induction of innate and adaptive immunity. Salmonella enterica is an important bacterial pathogen that causes disease in humans and livestock species. Transmission of some serovars (e.g. Enteritidis and Typhimurium) to humans is in the form of a food-borne zoonosis. To combat human gastroenteritis caused by salmonellae, vaccination of livestock animals to remove the major source of this organism is a more suitable approach than vaccination of people. Current vaccines are not particularly effective so understanding how to induce the most appropriate host immune responses to drive protective anti-Salmonella immunity is critical to optimise vaccination strategies against Salmonella spp. Salmonella can be located in intracellular (in vacuoles) and extracellular compartments but it also uses specialist structures to translocate products directly into the host cell cytoplasm. Hence, molecules derived from Salmonella are available to both the TLR and NLR pathways. PRR biology in domestic animals is in its infancy whereas PRR biology and the tools available to study these receptors in mice are well developed. In this project we will define which PRRs are important in controlling bacterial growth and mounting a protective immune response against infection with S. Typhimurium by infecting mouse strains with engineered deficiencies in the PRR pathways. The impact of this work will be to clearly define which PRRs drive protective anti-Salmonella immunity.

Publications

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Bryant CE (2010) The molecular basis of the host response to lipopolysaccharide. in Nature reviews. Microbiology

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Kenny EF (2009) MyD88 adaptor-like is not essential for TLR2 signaling and inhibits signaling by TLR3. in Journal of immunology (Baltimore, Md. : 1950)

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Man SM (2014) Actin polymerization as a key innate immune effector mechanism to control Salmonella infection. in Proceedings of the National Academy of Sciences of the United States of America

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Man SM (2014) Inflammasome activation causes dual recruitment of NLRC4 and NLRP3 to the same macromolecular complex. in Proceedings of the National Academy of Sciences of the United States of America

 
Description Salmonella enterica causes a wide range of diseases in many animals. Economic losses to the farming industry through Salmonella infection are potentially very high, but also important is the fact that a number of serovars that infect animals can also cause food poisoning and gastroenteritis in humans. Vaccination of animals to lower the levels of Salmonella in meat and eggs will reduce the chances of infection spreading to people. A successful vaccine must generate a strong adaptive immune response but must also be safe for application in young and older individuals. Understanding how Salmonella interacts with its host to generate an immune response is critical when trying to design new vaccines. Animals detect the presence of molecules on the bacteria through specialised proteins called Pattern Recognition Receptors (PRRs) which then initiate host defence mechanisms to clear the infection. These receptors are also believed to be important in generating a good response to vaccines, but precisely how this happens is currently unclear. In this grant we aimed to determine which PRRs are important for initiating bacterial clearance and for driving adaptive immune responses against Salmonella infections. The following key findings were made during this project: 1. The PRR Toll-like receptor 4 (TLR4) plays an important role in clearance of, and the induction of effective adaptive TH1 immune responses against, Salmonella Typhimurium. The TH1 response developed by cells without TLR4 is markedly reduced because of defective dendritic cell function. 2. The PRRs Nod-like receptors NLRP3 and NLRC4, through activation of the effector protein casapse 1, contribute to controlling the numbers of salmonellae. 3. Activation of NLRC4 dampens the adaptive TH1 immune responses against Salmonella Typhimurium. NLRC4 dampens TH1 immunity independently of caspase-1 identifying a novel mechanism by which anti-Salmonella adaptive immune responses are controlled. Our work suggests that PRR activity makes an important contribution to the immune responses generated against Salmonella. Some PRRs enhance adaptive immunity whilst others suppress it therefore these data will allow us to design better vaccines for chickens, other domestic animals and humans.
Exploitation Route 1. Generation of an optimal live attenuated vaccine 2. Data have been used for multiple presentations to the public to increase understanding of how animals detect bacteria. This work has clarified how PRRs contribute to the protective immune response against Salmonella infections. The information gained in this grant will allow us to design genetically modified salmonellae to give a live attenuated vaccine with modified bacterial PRR ligands such that the maximum protective immunity against this bacterum can be achieved.
Sectors Education,Pharmaceuticals and Medical Biotechnology

URL http://www.vet.cam.ac.uk/directory/ceb27@cam.ac.uk
 
Description Our findings have generated impact in several ways 1. Many of the observations made during the grant were used in communications to both the scientific community and the general public 2. Our findings were published in scientific journals 3. Our findings were the basis of two follow up grants: one to further understand how Salmonella vaccines may be generated for chickens and as a basis for an BBSRC Research Development fellowship to Bryant to develop collaborations with mathematicians and physicists 4. We hosted several school children in the lab teaching them about Salmonella.
First Year Of Impact 2009
Sector Agriculture, Food and Drink,Education
Impact Types Societal,Economic

 
Description A mathematical and biophysical analysis of salmonella-macrophage interactions
Amount £255,019 (GBP)
Funding ID BB/H021930/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 10/2010 
End 09/2013
 
Description Effects of Nod-like receptor activity on protective immunity against Salmonella infection
Amount £917,208 (GBP)
Funding ID BB/K006436/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 01/2013 
End 12/2016
 
Description Elion and Black Immunology Catalyst Sabbatical Award
Amount £226,000 (GBP)
Organisation GlaxoSmithKline (GSK) 
Sector Private
Country Global
Start 11/2016 
End 10/2019
 
Description Royal Society Wolfson Refurbishment Scheme
Amount £267,000 (GBP)
Organisation The Royal Society 
Sector Academic/University
Country United Kingdom
Start 03/2016 
End 02/2018
 
Description Continued Collaboration with University of Massachusetts Medical Schoo 
Organisation University of Massachusetts
Department University of Massachusetts Medical School
Country United States 
Sector Academic/University 
PI Contribution Continuation of existing collaboration
Collaborator Contribution Provision of cell lines and reagents. Intellectual discussions about our research work.
Impact On going provision of reagents has enabled our publications for this grant.
Start Year 2009
 
Description GSK Immunology Catalyst Sabbatical Award 
Organisation GlaxoSmithKline (GSK)
Country Global 
Sector Private 
PI Contribution GSK have bought out my teaching and administration duties for 3 years. I spend 3 days a week at GSK and have 2 postdoctoral researchers at GSK. I am working on inflammasome biology and Pattern Recognition Receptor research. I intellectually input into several of the GSK therapeutic units.
Collaborator Contribution GSK bought out my teaching and administration duties for 3 years and fund 2 postdoctoral researchers to work with me at GSK
Impact None yet
Start Year 2016
 
Description Genentech Visiting Professorship 
Organisation Genetech, Inc
Country United States 
Sector Private 
PI Contribution I visited Genentech South San Francisco site and performed research in the laboratory there for 9 months. Two of my PhD students also had 3 month internships working with myself and my Genentech collaborators. We learnt new techniques and gained access to new reagents. I intellectually contributed to their inflammasome research program.
Collaborator Contribution My collaborators gave me novel reagents and supported my research during my time in the USA
Impact None yet as the work is still in progress
Start Year 2016
 
Description Luke O'Neill, Trinity College, Dublin 
Organisation Trinity College Dublin
Country Ireland 
Sector Academic/University 
PI Contribution Collaboration with Trinity College, Dublin
Collaborator Contribution Intellectual input Collaborative discussion Transfer of new techniques
Impact Publications
Start Year 2007
 
Company Name Polypharmakos 
Description Spin out company with Monique Simmonds (Kew), Mark Holmes (Cambridge), Duncan Maskell (Cambridge) to screen natural products for antimicrobial and innate immune activity. 
Year Established 2016 
Impact None Yet
Website http://www.polypharmakos.com/
 
Description ANU, Canberra, Australia, "Pattern Recognition Receptor Signalling in response to Infection" 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact Talk to faculty and students at ANU with associated questions
Year(s) Of Engagement Activity 2018
 
Description Cambridge Science Fair Panel Discussion 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact I chaired a session and talks and a question and answer session on receptors at the Cambridge Science Fair
Year(s) Of Engagement Activity 2018
 
Description Discussion slot on the Naked Scientists Radio Show 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact A media interview where different aspects of immunology were reviewed and discussed
Year(s) Of Engagement Activity 2018
 
Description EMBO Inflammasome meeting Munich "Bacterial recognition by Pattern Recognition Receptors" 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact Talk at the EMBO international inflammasome meeting and associated questions
Year(s) Of Engagement Activity 2018
 
Description Hudson Innate Immunity Institute, Melbourne, Australia "Pattern Recognition Receptor Signalling in response to Infection" 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact Talk to faculty and students at the Hudson Institute and associated questions
Year(s) Of Engagement Activity 2018
 
Description International Cytokine Meeting, Boston, USA "Bacterial recognition by Pattern Recognition Receptors" 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact Talk at the international cytokine meeting and questions after.
Year(s) Of Engagement Activity 2018
 
Description Lafferty Debate participant 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact Participated in a public debate on "Adaptive Immunity is innately redundant"
Year(s) Of Engagement Activity 2016
 
Description Laser tweezers picking up bacteria 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach regional
Primary Audience Public/other audiences
Results and Impact Interview on the Naked Scientists Radio Show, Radio Cambridgeshire

no actual impacts realised to date
Year(s) Of Engagement Activity 2010
URL http://www.thenakedscientists.com/HTML/content/interviews/interview/1358/
 
Description NIH, Bethesda, USA "Pattern Recognition Receptor Signalling in response to Infection" 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Talk to staff and researcher at the NIH and responded to the associated questions
Year(s) Of Engagement Activity 2018
 
Description Pattern recognition receptors: the key to host recognition of infection or a load of old PAMPs 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Participants in your research and patient groups
Results and Impact Invited talk

no actual impacts realised to date
Year(s) Of Engagement Activity 2009
 
Description Pattern recognition receptors: the key to host recognition of infection or a load of old PAMPs 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Participants in your research and patient groups
Results and Impact Invited talk Institute for Child Health

no actual impacts realised to date
Year(s) Of Engagement Activity 2010
 
Description Pattern recognition receptors: the key to host recognition of infection or a load of old PAMPs 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Participants in your research or patient groups
Results and Impact Invited talk

no actual impacts realised to date
Year(s) Of Engagement Activity 2009
 
Description Pattern recognition receptors: the key to host recognition of infection or a load of old PAMPs 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Participants in your research and patient groups
Results and Impact Invited talk

no actual impacts realised to date
Year(s) Of Engagement Activity 2010
 
Description Pattern recognition receptors: therapeutic targets for allergic as well as infectious diseases 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Participants in your research or patient groups
Results and Impact Invited talk CIMR

no actual impacts realised to date
Year(s) Of Engagement Activity 2011
 
Description Recognition of bacterial infection by Pattern recognition receptors 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Participants in your research or patient groups
Results and Impact Invited talk NIAID/NIH

no actual impacts realised to date
Year(s) Of Engagement Activity 2011
 
Description Recognition of bacterial infection by Pattern recognition receptors 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Participants in your research or patient groups
Results and Impact Invited talk

no actual impacts realised to date
Year(s) Of Engagement Activity 2011
 
Description Talk Young Microbiologists Belfast 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Postgraduate students
Results and Impact Talk to young microbiologists (post graduate and post doctoral researchers) and associated questions
Year(s) Of Engagement Activity 2018
 
Description The Lorne Innate Immunity Meeting, Lorne, Australia "Pattern Recognition Receptor Signalling in response to Infection" 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact Talk to the iInnate Immunity meeting and the associated questions
Year(s) Of Engagement Activity 2018
 
Description The University of Brisbane, Australia "Pattern Recognition Receptor Signalling in response to Infection" 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact Talk to faculty and students at the The University of Brisbane with associated questions
Year(s) Of Engagement Activity 2018
 
Description There are 100 trillion Bacteria in Your Gut: How do we protect ourselves against infection? 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Schools
Results and Impact Lecture to year 12 school children Public lecture including research funded by this BBSRC grant

no actual impacts realised to date
Year(s) Of Engagement Activity 2013
 
Description There are 100 trillion Bacteria in Your Gut: How do we protect ourselves against infection? 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Talk to public at Queens' College Academic Saturday alumni event Public communication on how bacteria are recognised by the host: includes the research work on this BBSRC funded project

no actual impacts realised to date
Year(s) Of Engagement Activity 2012
 
Description There are 100 trillion Bacteria in Your Gut: How do we protect ourselves against infection? 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Public talk to the Society of Biology Public lecture to the Society for Biology in Cambridge

no actual impacts realised to date
Year(s) Of Engagement Activity 2013
 
Description Toll2018, Porto, Portugal "Bacterial recognition by Pattern Recognition Receptors" 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact Talk at the international Toll receptor meeting and associated questions
Year(s) Of Engagement Activity 2018
 
Description Trinity College Dublin "Pattern Recognition Receptor Signalling in response to Infection" 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact Talk to the faculty members and students at Trinity College Dublin plus associated questions
Year(s) Of Engagement Activity 2018
 
Description University of Baltimore "Pattern Recognition Receptor Signalling in response to Infection" 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact Talk to faculty and students at the University of Baltimore and responded to associated questions
Year(s) Of Engagement Activity 2018
 
Description University of Maryland, USA: Pattern recognition receptors: the key to host recognition of infection or a load of old PAMPs 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact Invited talk at University of Maryland, USA

no actual impacts realised to date
Year(s) Of Engagement Activity 2011
 
Description University of Strasbourg, France "Bacterial recognition by Pattern Recognition Receptors" 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact Talk to postgraduate students at the University of Strasberg
Year(s) Of Engagement Activity 2018
 
Description University of Trondheim, Norway "Bacterial recognition by Pattern Recognition Receptors" 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact Talk (and answered questions in response to talk) to students and researchers at the University of Trondheim in Norway.
Year(s) Of Engagement Activity 2018
 
Description Walter and Eliza Hall Institute for Medical Research, Melbourne, Australia "Pattern Recognition Receptor Signalling in response to Infection" 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact Talk to faculty members and students at Walter and Eliza Hall Institute for Medical Research with associated questions
Year(s) Of Engagement Activity 2018
 
Description What can maths and physics do for infection biology? 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact Invited talk

no actual impacts realised to date
Year(s) Of Engagement Activity 2012