Rapid typing of carbohydrate receptors of influenza viruses pathogenic for humans

Lead Research Organisation: Imperial College London
Department Name: Dept of Medicine

Abstract

We are trying to find out which structures in the human respiratory airways influenza viruses bind to. We hope to be able to identify avian H5N1 viruses that could cause a human pandemic.

We know that to start infection, influenza viruses first need to bind to carbohydrate structures (receptors) on cells. Normally, human and avian viruses bind to different carbohydrates.

When the genes of avian viruses change or become mixed with genes of human viruses, they can switch to have preference for human cells. Human pandemic viruses can emerge in this way.

It is very challenging to detect and determine particular carbohydrate structures that the viruses bind to. This is because carbohydrates on cells are very diverse. They are like a forest containing innumerable trees with only a few specimens of each.

We have been developing a unique approach to identifying receptors among this vast range of carbohydrates. This is Carbohydrate Microarray Technology that involves spotting minute amounts of carbohydrates from cells onto a glass slide.

We are adapting the microarray technology to detect binding of the proteins known as haemagglutinins of different influenza viruses. We hope by this approach to be able to predict the pandemic potential of a given virus.

Technical Summary

The emerging pandemic of influenza A H5N1 in feral and domestic avian species has affected some 30 countries to date from eastern Asia to Western Europe and northern Africa. The 169 confirmed human cases, with 91 fatalities, in 7 countries (to 13/02/06) emphasize the potential to cause a devastating pandemic. It is a high priority therefore to understand the mechanisms of emergence and risk of pandemic. The initial step in infection by the influenza virus is attachment of the virus particle to the surface of target cells through the interaction of the viral surface protein, haemagglutinin, with cell surface oligosaccharides that have terminal sialic acids. Understanding details of the oligosaccharide-binding specificities of influenza A viruses that infect humans and avians and have different tissue targets in the human will provide information crucial to gauging the pandemic risk posed by H5N1 and other subtypes. This project is focused on characterizing the range of host cell oligosaccharides that are attachment sites for prevailing and emergent influenza viruses that are pathogenic in the human in contradistinction to those of the avian and other mammalian influenza viruses that are not readily transmissible from an animal host to the human or from one person to another. Our laboratory has pioneered a carbohydrate microarray technology, and we have shown that it can be used for rapid screening of the interactions of soluble proteins, as well as virus particles, with the carbohydrate chains of glycoproteins and glycolipids. Coupled with mass spectrometry it is a validated approach for assigning novel carbohydrate ligands. Moreover our technology is unique in that it has provision for generating ?designer? microarrays from targeted tissues and secretions. Thus the specificities of binding (or adhesive footprints) of viruses can be determined for carbohydrate sequences that occur in the target host organs. Our aim is, in collaboration with Dr Alan Hay (World Influenza Centre NIMR) and Drs Steven Wharton and John Skehel (Virology Division NIMR), to apply our microarray technology with advanced mass spectrometry to characterize the distinctive carbohydrate sequences in human respiratory tract that are bound by influenza A viruses that are pathogenic in the human. An ultimate objective is to produce a microchip of arrayed oligosaccharide probes for the rapid typing of the binding ?footprints? of influenza virus isolates in order to survey for those that have preferred tropisms for the human and thus are a potential epidemiological hazard.

Publications

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Campanero-Rhodes MA (2007) N-glycolyl GM1 ganglioside as a receptor for simian virus 40. in Journal of virology

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Campanero-Rhodes MA (2006) Carbohydrate microarrays reveal sulphation as a modulator of siglec binding. in Biochemical and biophysical research communications

 
Description Our Nature Biotchnolgy paper on receptor binding by the pandeminc H1N1 2009 influenza virus was tabled for information at a September 2009 meeting of the Scientific Advisory Group for Emergencies (SAGE) in Whitehall
Geographic Reach National 
Policy Influence Type Participation in advisory committee
Impact Discussions are under way with HPA regarding our carrying out receptor-binding analyses on pandemic H1N1 2009 influenza viruses that have caused fatal infaction
 
Description Platform comprisons of carbohdrayte microarrays
Geographic Reach North America 
Policy Influence Type Participation in advisory committee
Impact Our carbohydrate microarray system is internationally recognized as making an impact in the unravelling of protein carbohydrate interactions of medical and biological importance. It is acknowledged as having a sufficient number of desirable features in the field, and discussions are under way to set up a formal comparative study of our system with that of the NIH sponsored US Consortium of Functional Glycomics.
 
Description 'Exploitation of Glycoarrays - Translation to End-Use'
Amount £149,400 (GBP)
Funding ID EP/G037604/1 
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Academic/University
Country United Kingdom
Start 01/2009 
End 12/2010
 
Description BBSRC Project Grant (Novel Fluorescent Oligosaccharide Probes for Quantitative Microarray Analyses of Carbohydrate-Protein Interactions)
Amount £98,633 (GBP)
Funding ID BB/G000735/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 10/2008 
End 09/2009
 
Description BBSRC Project Grant (Trehalose signalling: understanding and exploiting an emerging small molecule carbonhydrate paradigm)
Amount £320,840 (GBP)
Funding ID BB/D524240/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 01/2006 
End 03/2011
 
Description Immunogenic sugar moieties of prostate cancers (To NIH Call for 'Alliance of Glycobiologists for Detection of Cancer and Cancer Risk U01')
Amount £220,110 (GBP)
Organisation National Institutes of Health (NIH) 
Sector Public
Country United States
Start 08/2007 
End 07/2012
 
Description Studies of host cell carbohydrate receptors for adhesion proteins of Toxoplasma gondii
Amount £125,550 (GBP)
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 09/2007 
End 08/2010
 
Description Wellcome Trust Project Grant (Changes in receptor usage by influenza A (H5N1) viruses during infection of humans - monitoring pandemic potential using oligosaccharide microarrays from human airway tissues)
Amount £463,845 (GBP)
Funding ID 085572/A/08/Z 
Organisation Wellcome Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 06/2008 
End 05/2011
 
Title Carbohydrate microarray system 
Description Approximately 660 oligosaccharide probes robotically arrayed on nitocellulose coated glass slides 
Type Of Material Technology assay or reagent 
Year Produced 2008 
Provided To Others? Yes  
Impact A state-of-the-art carbohydrate microarray system. This is one of the two most comprehensive carbohydrate microarray systems in the world and it is the most diverse on account of its high content of natural oligosaccharide sequences that are difficult to synthesize and it is intended to encompass entire glycomes i.e. the carbohydrate repertoires of whole organisms. 
 
Description Glycan microarrays for bacterial adhesion studies 
Organisation Finnish Red Cross Blood Service
Country Finland 
Sector Public 
PI Contribution Preparation of oligosaccharide probes and fabrication of oligosaccharide microarray for bacterial adhesion studies. The on-going work also involves data analysis and presentation
Impact Led to a collaborative Research Contract; On going
Start Year 2009
 
Description Immunogenic sugar moieties of prostate cancers 
Organisation Stanford University
Country United States 
Sector Academic/University 
PI Contribution Our team is the analytical compnent of the this internaional collaboration
Impact Led to a NIH grant (NIH Call for 'Alliance of Glycobiologists for Detection of Cancer and Cancer Risk U01') Our team is carrying out analyses to define a prostate cancer-associated carbohydrate antigen
Start Year 2007
 
Description Studies of host cell carbohydrate receptors for adhesion proteins of Toxoplasma gondii 
Organisation Imperial College London
Country United Kingdom 
Sector Academic/University 
PI Contribution We have identifed by carbohydrate microarray analyses host-cell receptors for the apicomplexan parasite Toxoplasma gondii which infects warm blooded animals including humans, and for Neospora caninum which infects dogs and cattle
Impact Led to a BBSRC grant; On going; Three publications so far: 17491595 19750531 19593815
Start Year 2007
 
Description Trehalose signalling 
Organisation Rothamsted Research
Country United Kingdom 
Sector Academic/University 
PI Contribution To analyse, qualititively and quantitatively, synthetic probes of the unsual non-reducing sugar trehalose, and to set up carbohdrydrate microarrays of trehalose-containing sugars for microarray analyses of trehalose-binding signalling proteins
Impact Led to a BBSRC's 'SCIBS Initiative' Research Grant; ongoing
Start Year 2006
 
Description UK Glycoarrays Consortium 
Organisation University of Manchester
Country United Kingdom 
Sector Academic/University 
PI Contribution Our team is one of seven in the UK that applied and gained a UK Research Councils basic Technology Grant 2004-2008 entitled 'Glycoarrays' - strategies for high-throughput analysis of the glycome (co-ordinated by the University of Manchester). This grant has provided strong support for our microarray-related outputs, and has led to a succsful applcation for the present follow-up collaborative grant 'Exploitation of Glycoarrays - Translation to End-Use'
Impact 19901027 19750531 19741625 19593815 19426131 19399183 19284289 18524852 17855525 17656321 17491595 17116483 16647038 16371356
 
Title Carbohdryate microarrays for studies of endogenous carbohydrate recogntion systems as well as pathogen host interatctions 
Description Optimization of carbohydrate (oligosaccharide) presentation on microarrays 
Type Support Tool - For Fundamental Research
Current Stage Of Development Initial development
Year Development Stage Completed 2009
Development Status Actively seeking support
Impact Currently this carbohdryate microarray sytem is the only one we are aware of that can discern the distinctive receptor binding property of the pandemic H1N1 2009 influneza virus compared with seasonal H1N1 virus. 
 
Description Press Relase from Imperial College Press Office highlighting our demonstration of the potential of the 2009 pandemic H1N1 influenza virus to bind to receptors deep in the lung 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Primary Audience Media (as a channel to the public)
Results and Impact Interviews with many newspaper reporters for example from France, Portugal , Turkey, USA, Canada, China, India and Pakistan

News Feature in Nature, recorded inertvew in 'The Voice of America', articles the London Times, Daily Mail, numerous newpapers overseas
Year(s) Of Engagement Activity 2009