Factor H binding protein and vaccine approaches
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Professor Christoph Tang's presentation at Meningitis Research Foundation's 2013 conference, Meningitis & Septicaemia in Children & Adults
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Factor H binding protein and vaccine approaches
- 1. Factor H binding protein Christoph Tang Sir William Dunn School of Pathology University of Oxford
- 2. TpmA - elicits killing +/- functional antibodies i) ii) iii) iv) v) vi) expressed by all strains sequence is conserved understand structure:function inactivate its function elicits herd immunity easy/cheap to manufacture
- 3. fHbp- a key meningococcal antigen TpmA The perfect meningococcal Antigen - elicits killing +/- functional antibodies i) ii) iii) iv) v) vi) expressed by all strains sequence is conserved understand structure:function inactivate its function elicits herd immunity easy/cheap to manufacture MimA Most interesting meningococcal Antigen vaccine antigen pathogenesis host susceptibility
- 4. fHbp- a key meningococcal antigen Tan L et al. N Engl J Med 2010
- 5. - expressed by all strains? Yes, for > 99% of UK MenB isolates 923 UK isolates ST-11 T366 frame shift cc162 A650 frame shift cc286 fHbp deletion 4 have frame shifts 3 have a deletion
- 6. - sequence conservation? 923 UK isolates V1 101 fHbps Family B V2 Family A Masignani et al., J Exp Med 2003 Fletcher et al., Infect Immun 2004 Brehony et al. Microbiology 2009 V3 many different fHbps
- 7. - sequence conservation? PorA fHbp dark red is most conserved blue is most variable going via grey many different fHbps
- 8. - sequence conservation? 923 UK isolates V1 101 fHbps Family B V2 Family A Masignani et al., J Exp Med 2003 Fletcher et al., Infect Immun 2004 Brehony et al. Microbiology 2009 V3 many different fHbps
- 9. - sequence conservation? Novartis Pfizer x1 fhbp + other antigens x2 lipidated fhbps 1 144 110 238 312 89 2 78 306 61 236 276 37 249 254 295 318 4 5 313 303 298 62 14 309 220 307 258 10 299 13 311 297 292 86 87 210 305 15 68 118 310 109 302 19 83 119 16 106 296 308 24 25 301 95 147 23 104 22 202 34 21 102 300 30 174 29 293 212 294 45 185 59 85 94 84 160 47 304 10 5 8 11 18 22 23 32 35 60 103 162 167 174 198 213 226 254 269 364 461 865 1157 41/44 N/A V1 V2 V3 0 10 20 30 40 50 60 70 80 90 100 110 2008 data Family B Family A 120 many different fHbps some immunological cross reactivity within families Lucidarme et al. Clin Vacc Immunol 2010 some frequent peptides/correlate with ST
- 10. - understand structure:function CFH SCR6/7 Glu 304 Lys 351 His 371 Glu 341 Glu 283 Mascioni et al., J Biol Chem 2009 Cantini et al., J Biol Chem 2009 fHbp Schneider et al. Nature 2009
- 11. - understand structure:function Ligand mimicry Schneider et al. Nature 2009
- 12. - understand structure:function GAGs* CRP C3b 1 CFH 2 3 4 5 6 7 GAGs* Sialic acid C3b/d GAGs* C3b 8 9 10 11 12 13 14 15 16 17 Negative complement regulator - co-factor for fI cleavage of C3b - accelerates decay of C3bBb - binds fB multiple partners, multiple roles GWAS highlights CFH and CFHRs CFHRs antagonise CFH Understand molecular basis of host susceptibility 18 19 20
- 13. - understand structure:function GAGs* CRP C3b 1 2 3 4 5 CFH 6 7 GAGs* Sialic acid C3b/d GAGs* C3b 8 9 CFHR3 10 11 12 CFHR1 13 14 15 CFHR4 16 17 CFHR2 CFH locus multiple partners, multiple roles GWAS highlights CFH and CFHRs CFHRs antagonise CFH Understand molecular basis of host susceptibility Davilla et al. Nat Genet 2010 18 19 20 CFHR5
- 14. - understand structure:function GAGs* CRP C3b 1 2 3 4 5 CFH 6 7 GAGs* Sialic acid C3b/d GAGs* C3b 8 9 CFHR3 10 11 12 CFHR1 13 14 15 CFHR4 16 17 18 19 20 CFHR2 CFHR5 CFH locus multiple partners, multiple roles GWAS highlights CFH and CFHRs CFHRs antagonise CFH Understand molecular basis of host susceptibility Goicoechea de Jorge et al., PNAS 2013
- 15. - understand structure:function GAGs* CRP C3b 1 2 3 4 5 6 7 GAGs* Sialic acid C3b/d GAGs* C3b 8 9 10 11 12 13 14 15 16 17 18 19 20 fHBP CFH CFHR3 CFHR1 CFHR4 CFHR2 CFH locus multiple partners, multiple roles GWAS highlights CFH and CFHRs CFHRs antagonise CFH Understand molecular basis of host susceptibility CFHR5
- 16. - inactivate function fHBP Hide immunogenic epitopes Less opportunity for ‘blocking’ antibodies SCR 6 and 7 Reduce immunogenicity
- 17. - inactivate function Amino acid substitutions generate non-functional fHbps Johnson et al. PLoS Pathog. 2012
- 18. - inactivate function Amino acid substitutions generate non-functional fHbps Schneider et al. Nature 2009 B V1 double Glu Beernik et al. J Immunol 2010 Johnson et al. PLoS Pathog 2012 Arg-Ser 8 Ala subs. V2 14 Ala subs. V3 9 Ala subs. Jongerius et al. PLoS Pathog 2013 Rossi et al. Vaccine 2013 (two subs.) A one sub. Johnson et al. PLoS Pathog. 2012
- 19. - inactivate function Host modification human mouse human mouse human mouse fHbp Murine SCR6/7 Human SCR6/7 - humanising the binding site insufficient to allow murine fH to bind fHbp - SCR 6 and 7 have distinct orientations in mfH and hfH Johnson et al. PLoS Pathog. 2012
- 20. - inactivate function X C3b Host modification C3b SAP/GAGs mfH mfH SAP/GAGs ? C3b ? SAP/GAGs extra hfH chimeric fH fHbp + fHbp Non-functional fHbp as or more immunogenic than w/t fHbps Johnson et al. PLoS Pathog. 2012 Beernik et al. J Immunol. 2011
- 21. - easy to manufacture Distribution of fHbps in MRF collection 2010-2012 (n=916) 10% 34% v1 56% v2 v3 no V2 fHbp containing vaccine
- 22. - easy to manufacture fHbp V2 is inherently unstable Trypsin digestion Differential Scanning calorimetry (DSC) V1 V2 V3 no V2 fHbp containing vaccine Crystal structure
- 23. - easy to manufacture fHbp V2 is inherently unstable Trypsin digestion Differential Scanning calorimetry (DSC) Wild-type M6 M5 M4a M4b no V2 fHbp containing vaccine stable V2 fHbps
- 24. - easy to manufacture fHbp V2 is inherently unstable 5 of 10 single amino acid substitutions de-stabilise the antigen M6 R85A L135A V136A A204P V211A E222A T225A E252A no V2 fHbp containing vaccine stable V2 fHbps non-functional stable V2 fHbps
- 25. - ? elicits herd immunity N. meningitidis Temp. (°C) CssA fHbp (alpha-2,3-sialyltransferase) 30 37 42 Temperature acts as a danger signal Lst RmpM ? less fHbp at lower temp in the nasopharynx ? less effect of vaccine on carriage Oriente et al., J Bact 2010 Loh et al. Nature 2013
- 26. - What next for fHbp? Vaccine enhanced immunogenicity cross variant/family protection non-functional fHbps - Serum bactericidal activity
- 27. - What next for fHbp? Vaccine enhanced immunogenicity cross variant/family protection non-functional fHbps Naturally occurring variants Seib et al., Infect Immun 2009 Jongerius et al., PLoS Pathog 2013 Hybrid fHbp Scarselli et al., Sci Trans Med 2011
- 28. - What next for fHbp? Vaccine enhanced immunogenicity cross variant/family protection non-functional fHbps
- 29. - What next for fHbp? Vaccine enhanced immunogenicity cross variant/family protection non-functional fHbps Host susceptibility fHbp:fH interactions fHbp:CFHR interactions interactions with polymorphic proteins
- 30. Susan Lea Steven Johnson Joe Caesar Phil Ward Rachel Everitt Martin Maiden Odile Harrison - University of Oxford Ray Borrow Jay Lucidarme - Meningococcal Reference Laboratory Matthew Pickering - Imperial College London Elena Goicoechea de Jorge Hayley Lavender Rachel Exley Ilse Jongerius Stijn van den Veen Edmund Loh
Notes de l'éditeur
- Binding at SCR67 and E283/304Previous work in our lab has determined the crystal structure of fHbp shown here in turquoise, green and yellow in complex with fH shown in blueWe have shown that fHbp binds to a specific region of fH with high affinity, There are amino acid residues in fHbp which appear to be crucial, Glutamate at 283 and 304As fHbp binds to fH with high affinity, we hypothesised that this could affect its immunogenicity as a vaccine by (1) hiding immunogenic epitopes within the binding site and (2) also result in the development of autoantibodies as the immune system sees fH in complex with a foreign antigenI thus generated modified fHbps with mutations at these critical residues to assess whether these potentially non-fH binding proteins are immunogenic
- Binding at SCR67 and E283/304Previous work in our lab has determined the crystal structure of fHbp shown here in turquoise, green and yellow in complex with fH shown in blueWe have shown that fHbp binds to a specific region of fH with high affinity, There are amino acid residues in fHbp which appear to be crucial, Glutamate at 283 and 304As fHbp binds to fH with high affinity, we hypothesised that this could affect its immunogenicity as a vaccine by (1) hiding immunogenic epitopes within the binding site and (2) also result in the development of autoantibodies as the immune system sees fH in complex with a foreign antigenI thus generated modified fHbps with mutations at these critical residues to assess whether these potentially non-fH binding proteins are immunogenic
- Binding at SCR67 and E283/304Previous work in our lab has determined the crystal structure of fHbp shown here in turquoise, green and yellow in complex with fH shown in blueWe have shown that fHbp binds to a specific region of fH with high affinity, There are amino acid residues in fHbp which appear to be crucial, Glutamate at 283 and 304As fHbp binds to fH with high affinity, we hypothesised that this could affect its immunogenicity as a vaccine by (1) hiding immunogenic epitopes within the binding site and (2) also result in the development of autoantibodies as the immune system sees fH in complex with a foreign antigenI thus generated modified fHbps with mutations at these critical residues to assess whether these potentially non-fH binding proteins are immunogenic
- Binding at SCR67 and E283/304Previous work in our lab has determined the crystal structure of fHbp shown here in turquoise, green and yellow in complex with fH shown in blueWe have shown that fHbp binds to a specific region of fH with high affinity, There are amino acid residues in fHbp which appear to be crucial, Glutamate at 283 and 304As fHbp binds to fH with high affinity, we hypothesised that this could affect its immunogenicity as a vaccine by (1) hiding immunogenic epitopes within the binding site and (2) also result in the development of autoantibodies as the immune system sees fH in complex with a foreign antigenI thus generated modified fHbps with mutations at these critical residues to assess whether these potentially non-fH binding proteins are immunogenic
- Binding at SCR67 and E283/304Previous work in our lab has determined the crystal structure of fHbp shown here in turquoise, green and yellow in complex with fH shown in blueWe have shown that fHbp binds to a specific region of fH with high affinity, There are amino acid residues in fHbp which appear to be crucial, Glutamate at 283 and 304As fHbp binds to fH with high affinity, we hypothesised that this could affect its immunogenicity as a vaccine by (1) hiding immunogenic epitopes within the binding site and (2) also result in the development of autoantibodies as the immune system sees fH in complex with a foreign antigenI thus generated modified fHbps with mutations at these critical residues to assess whether these potentially non-fH binding proteins are immunogenic