factor h binding protein and vaccine approaches
DESCRIPTION
Professor Christoph Tang's presentation at Meningitis Research Foundation's 2013 conference, Meningitis & Septicaemia in Children & AdultsTRANSCRIPT
Factor H binding protein
Christoph TangSir William Dunn School of PathologyUniversity of Oxford
- elicits killing +/- functional antibodies
i) expressed by all strainsii) sequence is conservediii) understand structure:function iv) inactivate its functionv) elicits herd immunityvi) easy/cheap to manufacture
TpmA
- elicits killing +/- functional antibodies
i) expressed by all strainsii) sequence is conservediii) understand structure:function iv) inactivate its functionv) elicits herd immunityvi) easy/cheap to manufacture
TpmA MimA
fHbp- a key meningococcal antigen
The perfect meningococcal Antigen Most interesting meningococcal Antigen
vaccine antigen
pathogenesis
host susceptibility
Tan L et al. N Engl J Med 2010
fHbp- a key meningococcal antigen
- expressed by all strains?
Yes, for > 99% of UK MenB isolates
ST-11 DT366 frame shiftcc162 DA650 frame shift
cc286 fHbp deletion
923 UK isolates
4 have frame shifts
3 have a deletion
- sequence conservation?
Masignani et al., J Exp Med 2003Fletcher et al., Infect Immun 2004Brehony et al. Microbiology 2009
Family A
Family B
V1
V2
V3many different fHbps
923 UK isolates
101 fHbps
- sequence conservation?
many different fHbps
PorA fHbpdark red is most conserved
blue is most variable going via grey
- sequence conservation?
Masignani et al., J Exp Med 2003Fletcher et al., Infect Immun 2004Brehony et al. Microbiology 2009
Family A
Family B
V1
V2
V3many different fHbps
923 UK isolates
101 fHbps
0 10 20 30 40 50 60 70 80 90 100 110 120
30447
16084948559
18545
294212293
29174
30300102
2134
20222
10423
14795
3012524
308296106
16119
8319
302109310118
6815
305210
8786
292297311
13299
10258307220309
1462
298303313
54
318295254249
37276236
61306
782
89312238110144
1
5811182223323560103162167174198213226254269364461865115741/44N/A
10
Family A
Family B
V1
V2
V3
- sequence conservation?Novartisx1 fhbp
+ other antigens
Pfizerx2 lipidated fhbps
many different fHbpssome immunological cross reactivity within families
some frequent peptides/correlate with STLucidarme et al. Clin Vacc Immunol 2010
2008 data
- understand structure:function
Schneider et al. Nature 2009Mascioni et al., J Biol Chem 2009Cantini et al., J Biol Chem 2009
Glu 341
Glu 304
His 371
Lys 351
Glu 283
CFH SCR6/7
fHbp
- understand structure:function
Ligand mimicry
Schneider et al. Nature 2009
- understand structure:function
1 2 53 4 6 7 108 9 11 12 1513 14 16 17 2018 19
C3bGAGs*
CRPGAGs*
C3b
GAGs*Sialic acid
C3b/d
CFH Negative complement regulator- co-factor for fI cleavage of C3b- accelerates decay of C3bBb- binds fB
multiple partners, multiple rolesGWAS highlights CFH and CFHRs
CFHRs antagonise CFHUnderstand molecular basis of host susceptibility
Davilla et al. Nat Genet 2010
CFH CFHR3 CFHR1 CFHR4 CFHR2 CFHR5
CFH locus
- understand structure:function
1 2 53 4 6 7 108 9 11 12 1513 14 16 17 2018 19
C3bGAGs*
CRPGAGs*
C3b
GAGs*Sialic acid
C3b/d
multiple partners, multiple rolesGWAS highlights CFH and CFHRs
CFHRs antagonise CFHUnderstand molecular basis of host susceptibility
CFH CFHR3 CFHR1 CFHR4 CFHR2 CFHR5
CFH locus
- understand structure:function
1 2 53 4 6 7 108 9 11 12 1513 14 16 17 2018 19
C3bGAGs*
CRPGAGs*
C3b
GAGs*Sialic acid
C3b/d
Goicoechea de Jorge et al., PNAS 2013
multiple partners, multiple rolesGWAS highlights CFH and CFHRs
CFHRs antagonise CFHUnderstand molecular basis of host susceptibility
CFH CFHR3 CFHR1 CFHR4 CFHR2 CFHR5
CFH locus
- understand structure:function
1 2 53 4 6 7 108 9 11 12 1513 14 16 17 2018 19
C3bGAGs*
CRPGAGs*
C3b
GAGs*Sialic acid
C3b/d
multiple partners, multiple rolesGWAS highlights CFH and CFHRs
CFHRs antagonise CFHUnderstand molecular basis of host susceptibility
fHBP
- inactivate function
fHBP
SCR 6 and 7
Hide immunogenic epitopes
Reduce immunogenicity
Less opportunity for ‘blocking’ antibodies
- inactivate function
Johnson et al. PLoS Pathog. 2012
Amino acid substitutions generate non-functional fHbps
- inactivate function
Schneider et al. Beernik et al. Johnson et al. Jongerius et al. Rossi et al. Nature 2009 J Immunol 2010 PLoS Pathog 2012 PLoS Pathog 2013 Vaccine 2013
V1 double Glu Arg-Ser 8 Ala subs.
V2 14 Ala subs. (two subs.)
V3 9 Ala subs. one sub.
B
A
Johnson et al. PLoS Pathog. 2012
Amino acid substitutions generate non-functional fHbps
Host modification- inactivate function
Johnson et al. PLoS Pathog. 2012
fHbp
Human SCR6/7
Murine SCR6/7
humanmouse
humanmousehumanmouse
- humanising the binding site insufficient to allow murine fH to bind fHbp- SCR 6 and 7 have distinct orientations in mfH and hfH
Host modification- inactivate function
fHbp
chimericfH
XC3b SAP/GAGs
Johnson et al. PLoS Pathog. 2012
C3b SAP/GAGsmfH
fHbp
? C3b ? SAP/GAGsextrahfH
+
Beernik et al. J Immunol. 2011
mfH
Non-functional fHbp as or more immunogenic than w/t fHbps
- easy to manufacture
no V2 fHbp containing vaccine
56%34%
10%
Distribution of fHbps in MRF collection 2010-2012 (n=916)
v1 v2 v3
Differential Scanning calorimetry (DSC)Trypsin digestion
V1
V2
V3
Crystal structure
- easy to manufacture
fHbp V2 is inherently unstable
no V2 fHbp containing vaccine
Wild-typeM6M5M4aM4b
fHbp V2 is inherently unstable
- easy to manufacture
Differential Scanning calorimetry (DSC)Trypsin digestion
no V2 fHbp containing vaccinestable V2 fHbps
fHbp V2 is inherently unstable
- easy to manufacture
no V2 fHbp containing vaccinestable V2 fHbps
non-functional stable V2 fHbps
M6R85AL135AV136AA204PV211AE222AT225AE252A
5 of 10 single amino acid substitutions de-stabilise the antigen
N. meningitidis
30 37 42
fHbp
Temp. (°C)
RmpM
CssA
Lst (alpha-2,3-sialyltransferase)
- ? elicits herd immunity
Temperature acts as a danger signal
Loh et al. Nature 2013Oriente et al., J Bact 2010
? less fHbp at lower temp in the nasopharynx? less effect of vaccine on carriage
Vaccine enhanced immunogenicitycross variant/family protectionnon-functional fHbps
- What next for fHbp?
- Serum bactericidal activity
Naturally occurring variants
Seib et al., Infect Immun 2009Jongerius et al., PLoS Pathog 2013
Hybrid fHbp
Scarselli et al., Sci Trans Med 2011
- What next for fHbp?
Vaccine enhanced immunogenicitycross variant/family protectionnon-functional fHbps
- What next for fHbp?
Vaccine enhanced immunogenicitycross variant/family protectionnon-functional fHbps
- What next for fHbp?
Vaccine enhanced immunogenicitycross variant/family protectionnon-functional fHbps
Host susceptibility fHbp:fH interactionsfHbp:CFHR interactionsinteractions with polymorphic
proteins
Susan Lea - University of OxfordSteven JohnsonJoe CaesarPhil WardRachel Everitt
Martin MaidenOdile Harrison
Ray Borrow - Meningococcal Reference LaboratoryJay Lucidarme
Matthew Pickering - Imperial College LondonElena Goicoechea de Jorge
Stijn van den Veen
Rachel Exley
Edmund Loh
HayleyLavender
Ilse Jongerius