1. HIV broadly neutralizing antibodies: learning lessons from infections
Penny Moore
National Institute for Communicable Diseases, a division of the
National Health Laboratory Service of South Africa,
University of the Witwatersrand, Johannesburg, South Africa
and the Centre for the AIDS Program of Research in South Africa (CAPRISA)
IAS, 2014, Melbourne

2. Broadly neutralizing antibodies (BNAbs) develop in a fifth of HIV infected people
Elite neutralizers (1%)
Screening of chronically infected people shows BCN antibodies develop fairly often...
Broad neutralizers (20%)
No/limited breadth

3. Isolation of monoclonal anti-HIV antibodies
We know (most of) the targets
V2/glycan
>12 mAbs
V3/glycan supersite
>25 mAbs
gp120-gp41 interface
>3 mAbs
CD4bs
>25 mAbs
MPER
>5 mAbs
Modified from Burton et al., Science 2012

4. Ontogeny of BNAbs: How do BNAbs develop?
Breadth
years of infection
UCA
(unmutated common ancestor)

5. Ontogeny of BNAbs: How do BNAbs develop?
Breadth
years of infection
UCA

6. Ontogeny of BNAbs: How do BNAbs develop?
Breadth
years of infection
UCA

7. Developmental pathways are likely to differ by epitope
V2/glycan
>12 mAbs
Long CDRH3 (>25 aa) to penetrate glycan shield
V3/glycan
>25 mAbs
Heavily mutated (30%)
CD4bs
>25 mAbs
MPER
>5 mAbs
Modified from Burton et al., Science 2012

8. Developmental pathways are likely to differ by epitope
V2/glycan
>12 mAbs
V3/glycan
>25 mAbs
Heavily mutated (30%)
CD4bs
>25 mAbs
MPER
>5 mAbs
Modified from Burton et al., Science 2012

9. CD4bs BNAbs – highly affinity matured
Unmutated common ancestor
Mature BNAb
Highly mutated away from their ancestor
years of infection

10. Do long CDR H3s similarly develop gradually?
V2/glycan
>12 mAbs
Long CDRH3 (>25 aa) to penetrate glycan shield
V3/glycan
>25 mAbs
MPER
>5 mAbs
Modified from Burton et al., Science 2012

11. CAP256 targets V2
CAP256
Gray et al 2011, Moore et al 2011

12. Twelve related mAbs isolated by B cell culture
V1V2-directed
NAbs
Neutralization
breadth starts
Weeks
post-infection *
Week
Antibody
Somatic mutations (nt)
CDR H3 length (aa)
Neutralization (46 strains)
Vl1-51*02
VH3-30*18
Breadth
Potency
(IC50, μg/ml) 59
CAP256-VRC26.01
3.9%
8.3% 35
15%
2.93
119
CAP256-VRC26.02
4.9%
8.7%
17%
0.40
CAP256-VRC26.03
7.4%
30%
0.06
CAP256-VRC26.04
8.1%
9.0%
28%
0.25
CAP256-VRC26.05
5.4%
10.1%
22%
0.10
CAP256-VRC26.06
10.8% 36
0.16
CAP256-VRC26.07
7.7%
11.8%
13%
2.02
CAP256-VRC26.08
9.8% 37
46%
0.14
CAP256-VRC26.09
14.2%
0.08
206
CAP256-VRC26.10
24%
0.60
CAP256-VRC26.11
13.7%
11.9%
0.82
CAP256-VRC26.12
8.4%
15.3% 7%
0.49
Doria-Rose, Schramm, Gorman, Moore et al, Nature, 2014

13. CAP256-VRC26 has typical features of anti-V2 broad neutralizing antibodies
PG9
CAP256-VRC26
Antibody
BG505
SOSIP
HIV Env
Jason Gorman and Peter Kwong
HL Kim, A Cupo, R Sanders, IA Wilson, JP Moore, AB Ward
Ideal for defining the developmental pathways of V2 antibodies with long CDR H3

14. CAP256-VRC26 heavy chain and light chain antibody sequences
VH3-30*18
VL1-52*02
UCA_H
UCA_L
CAP256-VRC26.01
CAP256-VRC26.10 CAP256-VRC26.11
CAP256-VRC26.06
CAP256-VRC26.05
CAP256-VRC26.02
CAP256-VRC26.04
CAP256-VRC26.03
CAP256-VRC26.07
CAP256-VRC26.12
CAP256-VRC26.09
CAP256-VRC26.08
The UCA had a 35 amino acid CDR H3 fully formed through VDJ recombination.
Heavy Chain longitudinal phylogenetic tree
Light Chain longitudinal phylogenetic tree
Week 38 48 59
119
176
206
Evolutionary distance
0.02
Chaim Schramm and Larry Shapiro

15. Unmutated common ancestor
The CAP256-VRC26 unmutated common ancestor had a fully formed long CDR H3
Long CDRH3s
Mature BNAb
Unmutated common ancestor

16. Unmutated common ancestor
The CAP256-VRC26 unmutated common ancestor had a fully formed long CDR H3
Long CDRH3s
Was this enough for breadth?
Mature BNAb
Unmutated common ancestor

17. Development of CAP256-VRC26.01
V1V2-directed
NAbs
Neutralization
breadth starts
Weeks
post-infection *
Week
Antibody
Somatic mutations (nt)
CDR H3 length (aa)
Neutralization (46 strains)
Vl1-51*02
VH3-30*18
Breadth
Potency
(IC50, μg/ml) 59
CAP256-VRC26.01
3.9%
8.3% 35
15%
2.93
119
CAP256-VRC26.02
4.9%
8.7%
17%
0.40
CAP256-VRC26.03
7.4%
30%
0.06
CAP256-VRC26.04
8.1%
9.0%
28%
0.25
CAP256-VRC26.05
5.4%
10.1%
22%
0.10
CAP256-VRC26.06
10.8% 36
0.16
CAP256-VRC26.07
7.7%
11.8%
13%
2.02
CAP256-VRC26.08
9.8% 37
46%
0.14
CAP256-VRC26.09
14.2%
0.08
206
CAP256-VRC26.10
24%
0.60
CAP256-VRC26.11
13.7%
11.9%
0.82
CAP256-VRC26.12
8.4%
15.3% 7%
0.49
Nicole Doria-Rose, Ryan Staupe, Nancy Longo, Jinal Bhiman

18. Rapid development of neutralization breadth within the CAP256-VRC26 lineage
VRC26-UCA
VRC26-I1
VRC26-I2
VRC26.01
Chaim Schramm

19. Rapid development of neutralization breadth within the CAP256-VRC26 lineage
VRC26-UCA
VRC26-I1
VRC26-I2
VRC26.01
CAP256 SI
The CAP256-VRC26 UCA, despite having a long CDR H3, can ONLY neutralize the virus that infected CAP no neutralization breadth

20. Rapid development of neutralization breadth within the CAP256-VRC26 lineage
VRC26-UCA
VRC26-I1
VRC26-I2
VRC26.01
CAP256 SII
Chaim Schramm

21. Only moderate levels of somatic hypermutation needed for breadth
Mature BNAb with moderate somatic hypermutation
Unmutated common ancestor
Weeks…. ✗
years of infection

22. Ontogeny of BNAbs: Looking backwards from breadth
years of infection
UCA

23. Dramatic viral changes immediately precede CAP256 neutralization breadth
Maturation of the anti-V2 lineage CAP256-VRC26 was associated with viral replacement in the epitope
Broadly neutralizing antibodies emerge in the context of multiple immunotypes created through viral diversification
V1V2 mutations
Duration of infection
CAP256-VRC26 emerges in peripheral B cell repertoire

24. Dramatic viral changes immediately precede CAP256 neutralization breadth
Maturation of the anti-V2 lineage CAP256-VRC26 was associated with viral replacement in the epitope
Broadly neutralizing antibodies emerge in the context of multiple immunotypes created through viral diversification
V1V2 mutations
Duration of infection
CAP256-VRC26 emerges in peripheral B cell repertoire
Plasma heterologous neutralization develops
Jinal Bhiman, Daniel Sheward, Molati Nonyane, Bronwen Lambson

25. Emergence of escape mutations drives increased breadth
166
CAP256 infecting virus
94 weeks p.i.
R166, a crucial part of the CAP256 epitope, undergoes selection pressure as bNAbs mature
Jinal Bhiman, Daniel Sheward

26. Emergence of new immunotypes drives increased breadth – K166
All subtype C viruses (n=1,005)
166
CAP256 infecting virus
166
94 weeks p.i.

27. Emergence of new immunotypes drives increased breadth – K166
All subtype C viruses (n=1,005)
166
CAP256 infecting virus
166
94 weeks p.i.
CAP256
CAP256 R166K
Percent breadth

28. Summary
Isolated PG9-type V1V2 neutralizing mAbs from seroconverting donor CAP256
CAP256 mAbs have a characteristic long anionic CDRH3 formed by initial gene recombination; i.e., present on naïve BCR
The UCA was capable of strain-specific neutralization, with modest affinity maturation conferring breadth
Viral diversification and co-evolution was associated with breadth, through bNAb tolerance of escape mutations
Doria-Rose, Schramm, Gorman, Moore et al, Nature, 2014

29. Which pathway is more amenable to HIV vaccine design?
CD4bs - highly mutated away from their ancestor
V1V2 - Long CDRH3s
Requires the engagement of a BCR with a long CDR H3 - these B cells are very rare
No requirement for long CDR H3, but Ig allele skewing may limit viable BCRs
Once stimulated, V1V2 BNAbs can develop within months, not years
May need high levels of affinity maturation take years – hard to achieve through vaccination
Immunogens may need to recreate antigenic diversity to drive affinity maturation
Immunogens may need to recreate antigenic diversity to drive affinity maturation

30. Acknowledgements NICD Jinal Bhiman IAVI NAC, CHAVI-ID and Scripps
Participant CAP256
Slim Abdool Karim, Quarraisha Abdool Karim, Nigel Garrett and the CAPRISA staff
NICD Jinal Bhiman
Kurt Wibner Molati Nonyane Bronwen Lambson
Thandeka Khoza Mashudu Madzivhandila
Lynn Morris
Mascola lab
Nicole Doria-Rose
R. Staupe, R. Roark M. Ernandes
Rebecca Lynch Rui Kong
Nancy Longo
Sijy O’Dell
Stephen Schmidt
Krisha McKee Mark Louder
John Mascola
Kwong lab
Jason Gorman Marie Pancera
Tongqing Zhou
Baoshan Zhang
Ivelin Georgiev
Young Do Kwon
Yongping Yang
Peter Kwong
Bioinformatics
Chaim Schramm
Zhenhai Zhang
Cinque Soto Ivelin Georgiev
Larry Shapiro
IAVI NAC, CHAVI-ID and Scripps
Andrew Ward
Helen Kim
Albert Cupo John Moore Rogier Sanders Wayne Koff Ian Wilson
Dennis Burton
U. Texas Austin Brandon DeKosky George Georgiou
Williamson lab
Daniel Sheward
Carolyn Williamson
Torrey Pines Emma Crooks James Binley
NIH Sequencing Core Holly Coleman
Brian Schmidt
Morgan Park
Jim Mullikin
NIAID NVITAL
Ellen Turk
Bob Bailer