Factor H binding protein and vaccine approaches

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


many different fHbps

PorA fHbpdark red is most conserved

blue is most variable going via grey


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


Ligand mimicry

Schneider et al. Nature 2009


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


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 locus


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




CFH locus


1 2 53 4 6 7 108 9 11 12 1513 14 16 17 2018 19

C3bGAGs*

CRPGAGs*

C3b

GAGs*Sialic acid

C3b/d



fHBP

- inactivate function

fHBP

SCR 6 and 7

Hide immunogenic epitopes

Reduce immunogenicity

Less opportunity for ‘blocking’ antibodies


Johnson et al. PLoS Pathog. 2012

Amino acid substitutions generate non-functional fHbps


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


Amino acid substitutions generate non-functional fHbps

Host modification- inactivate function


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


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


fHbp V2 is inherently unstable

no V2 fHbp containing vaccine

Wild-typeM6M5M4aM4b



Differential Scanning calorimetry (DSC)Trypsin digestion

no V2 fHbp containing vaccinestable V2 fHbps



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





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

https://sites.google.com/site/tangslab/people/dr-ilse-jongerius/Ilse%20Jongerius.jpg?attredirects=0

Factor H binding protein and vaccine approaches

Health & Medicine

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