ICRES integrated Chikungunya Research

Saturday, 1/10/2016

ICRES is an acronym for Integrated Chikungunya Research and is a collaborative project supported by the European Union under the Health Cooperation Work Programme of the 7th Framework programme (grant agreement number 261202). This project will integrate the expertise of EU laboratories with a long and strong track record of research on alpha-virus with EU laboratories that started work on CHIKV following the outbreak in La Reunion and laboratories from South East Asia working on this virus. The project will generate new molecular and cellular tools for research and applied studies, and develop a vaccine which at the end of this project is ready to enter clinical trials.

Principle Objectives :

1. Generate new molecular and cellular tools for research and applied studies including high- throughput screening and vaccines
2. Standardise, quality assure and distribute key diagnostic tests and develop new ones
3. Determine key virus genetic changes across time, geographical regions and species
4. Discover interactions between virus and human cells to inform the rational design of therapeutics
5. Determine pathogenesis of the acute and chronic disease in humans, including whether virus persists in joints, the cell types involved and the relationship to immune responses
6. Characterise rodent and non-human primate models of acute and chronic infection to further study the pathogenesis and to provide models for antiviral and vaccine screens.
7. Screen libraries of small molecular weight compounds for antiviral activity
8. Develop a vaccine which at the end of this project is ready to enter clinical trial.

Background:
• Since 2005 Chikungunya fever has affected millions of people producing a high fever and a debilitating arthralgia which can persist for months and progress to chronic arthritis.
• The current epidemic rose to prominence in 2005/6 following infection of >250,000 people on La Reunion.
• The virus rapidly spread to other islands in the Indian Ocean, India and SE Asia.
• Chikungunya cases in returning travellers have been reported. In summer 2007 a traveler from India to Italy initiated a locally transmitted outbreak which included one death from encephalitis.
• The mosquitoes transmitting this infection are spreading and increasing in Europe and could spread as far north as the British Isles.

ICRES
• To integrate the expertise of EU laboratories with a long and strong track record of research on alpha-viruses with EU laboratories that started work on CHIKV following the 2005 outbreak in La Raunion (France) and laboratories from SE Asia working on this virus in the current epidemic in this region. The consortium has met every 6 months. The Partners and their research group members are working together as a team and all work packages have made good progress.
• An open international meeting, Chikungunya-2013 is being planned for October 2013. A new molecular clone of CHIKV LR2006-OPY1, ICRES1, has been generated and its phenotype verified in macaques. ICRES-1 derived molecular constructs expressing full length virus sequence, virus replicas or virus structural proteins, various mutations and vaccine candidates and additions to these, including biochemical and fluorescent markers have been engineered and tested in vitro, and in some cases in vivo. A virus replicon particle (VRP) system has been established.
• Antibodies to CHIKV have been generated as have cells lines inducible for the expression of CHIKV proteins and cells which persistently express the CHIKV replicase driving a marker gene. The latter has been used to establish a high throughput in vitro screening system for antivirals.

The above viruses and reagents have been disseminated around the consortium members and are being used in the next stages of the programme. Methods for diagnostics and key laboratory assays including, plaque assays, ELISPOT, FACS, ELISA and RT-PCR have been reviewed and standardised across the consortium and cell lines have been exchanged to further ensure standardisation. Studies have started on pathway biology analysis of the interaction of the virus with mammalian cells. This includes transcriptomics, a yeast-two-hybrid assay of virus and cell protein interactions, analysis of cellular proteins which interact with the virus replicase complex, a SiRNA knock down screen to identify important cellular determinates of virus replication and anti-viral screening against libraries of known compounds. The methodologies and optimised parameters for all of these assays have been established at the required scale and preliminary results obtained. A mouse model of infection has been established in three laboratories and in vivo imaging using biochemical (luciferase) marker virus and screening of vaccine candidates carried out. The macaque model has been refined by showing that it is possible to target CHIKV infection to wrist joint and thereby establish a site at which the course of infection and inflammation can be reproducibly studied and a longitudinal study of immune parameters of the acute infection has been carried out and the samples are being analysed.

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