Development of a predictive model for vaccine matching for serotype O FMDV from serology and capsid sequence
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1 Development of a predictive model for vaccine matching for serotype O FMDV from serology and capsid sequence D. Borley, S. Upadhyaya, D. Paton, R. Reeve and Mana Mahapatra Pirbright Laboratory United Kingdom
2 FMDV Genome Organization AUG AUG Lab Lb Cleavage sites L 3C 2A Unknown VPg 3B 1-3 L VP0 VP3 VP1 2A 2B 2C 3A 3C 3D AAA n P1-2A P2 P3 3B 1-3 VP0 VP3 VP1 2A 2B 2C 3A 3C 3D 3B 1-3 VP0 VP3 VP1 2A 2B 2C 3A 3C 3D 2B 2C 3A 3C 3D VP4 VP2 3A 3C 3D Structural Non-Structural Proteins Proteins Capsid RNA Synthesis/Replication, Helicase /polymerase/protease Antigenicity, receptor binding activity, virulence
3 Main Objectives - Develop a predictive model for vaccine matching for serotype O FMDV - Genetic and/or structural determinants of antibodymediated protection a) b) 5x To develop/improve FMDV vaccine strain selection, and design novel vaccine 3x 2x 3x VP1- blue VP2- red VP3- green
4 Selection of suitable vaccine strains Vaccination using killed viral capsid antigens is very important for disease control Seven serotypes and multiple subtypes of FMDV Antigenic mis-match is one important cause for vaccine failure Vaccine strains must be selected from the available pool and the need for new vaccines must be identified eg. muiltiple outbreak in S. Korea Current vaccine selection - VNT or ELISA tests to measure the cross-reaction of a bovine vaccinal sera with the field strain in question Few studies have been carried out to measure cross-protection directly SAT2 SAT1 C SAT3 Asia1 A O
5 Problems with serological methods Time needed to grow up field viruses and not all grow equally well Need for panels of vaccine strains and antisera Antisera are inherently variable Difficult to standardise tests or to have full confidence in results without many repetition Vaccine selection based on serological methods always not give same results as in vivo
6 Alternative approaches I. Matching ELISA using type specific monoclonal antibodies (mabs) -Need panel of well defined mabs able to recognise differences between vaccine and field viruses -Need to know which antigenic sites are the most important II. Antigenic Cartography (Mahapatra et al., 2008) III. Sequencing of viral capsid and correlation of amino acid changes to antigenic matching -Still need to know which sites are the most significant
7 Viruses used -serotype O EURO-SA Cathay Serotype O viruses (n = 80) Bovine serum (n = 5) Asia Africa
8 Heatmap of viruses/antisera
9 III- Correlating capsid sequence to serology 5 sera X~80 viruses - Capsid sequence determined - Surface accessibility of each capsid residue determined * 48 discrete regions (1-40 aa) * Optimised serology result * LME prediction model developed VNT ELISA O BFS Multimer: VP1- Red, VP2 Blue, VP3 - Green (Borley et al. - manuscript in prep.)
10 Prediction model using VNT and Capsid sequence For the first time serotype O vaccine strain can be selected without recourse to serology
11 Predicted regions using model VNT ELISA Blue: VP Green: VP Red: VP Blue: VP Green: VP Red: VP
12 Neu. Ab Titre Individual aa tested VNT ELISA VP2 191 VP1 138 VP1 174 VP2 194 Site 2 Site 1 VP2 191 VP1 198 VP1 138 VP3 219 O1 BFS Ab. repertoire of bovine sera Epitope prediction (structure) 300 O BFS O1K Red. O BFS Red. O1K A C SAT ASN 190 ASN 190 ASN 190 ASN 190 SER 190 ASN 190 ASP 190 THR 191 THR 191 THR 191 THR 191 ASN 191 THR 191 GLN 191 VP2 100 GLU 192 GLU 192 GLU 192 GLU 192 ALA 192 THR 192 ASP 69 ASP 69 ASP 69 ASP 69 ASP 69 SER 69 0 Site1 Site 2 Site3 Site 4 Site 5 mab escape mutants SER 70 ASP 70 GLY 70 ASP 71 ASP 71 ASP 71 ASP 71 THR 71 SER 71 ARG 218 ARG 218 GLU 221 GLN 219 ARG 220 ALA 219 GLU 220 GLN 220 GLN 221 VP3 (Mahapatra et al., 2012) (Borley et al., submitted)
13 Serum antibody titre Reverse genetics to test the residues Type O cdna clone Mutations introduced VP2 191 and VP3 219 Recombinant virus - recovered, characterised Antigenic properties of the virus - serology Parent VP3 219 VP2 191
14 Conclusion Serological methods- time-consuming, still useful mab-based assay- vaccine-sp, not feasible (limited resources) Sequence vs serology model: -promising, needs further work - better rep. sera/viruses - testing and validation - may change in future VNT and ELISA different results (aa) New epitope VP2 191, VP3 219
15 What we would like to do next Refine the prediction model (type O): -Include additional sera against antigenically distant vaccine -Develop further model to confirm the residues predicted in the current model Test and validate the type O model: - Introduce in to use by FMD Ref. labs - Cross-protection studies Test the predicted aa residues using a cdna clone Extend the work to serotype A
16 Pirbright Daryl Borley Sasmita Upadhyaya Amin Asfor Fufa Bari David Paton Jef Hammond Acknowledgements Univ. of Glasgow Richard Reeve Dan Haydon Oxford Elizabeth Fry Dave Stuart Nico Visser Danny Gooverts IZSLER, Brescia Emi Brocchi Santina Grazioli
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