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

big

bigger

biggest

Broadly neutralizing antibodies (BNAbs) develop in a fifth of HIV infected people

Elite neutralizers (1%)

Broad neutralizers (20%)

No/limited breadth

Screening of chronically infected people shows BCN antibodies develop fairly often...

V2/glycan>12 mAbs

V3/glycan supersite>25 mAbs

CD4bs

>25 mAbs MPER>5 mAbs

Modified from Burton et al., Science 2012

Isolation of monoclonal anti-HIV antibodiesWe know (most of) the targets

gp120-gp41

interface>3 mAbs

Ontogeny of BNAbs: How do BNAbs develop?

years of infection

Breadth

UCA(unmutated

common ancestor)

years of infection

Breadth

UCA

Ontogeny of BNAbs: How do BNAbs develop?

years of infection

Breadth

UCA

Ontogeny of BNAbs: How do BNAbs develop?

V2/glycan>12 mAbs

V3/glycan>25 mAbs

CD4bs

>25 mAbs MPER>5 mAbs

Modified from Burton et al., Science 2012

Long CDRH3 (>25 aa) to penetrate glycan shield

Heavily mutated (30%)

Developmental pathways are likely to differ by epitope

V2/glycan>12 mAbs

V3/glycan>25 mAbs

CD4bs

>25 mAbs MPER>5 mAbs

Modified from Burton et al., Science 2012

Heavily mutated (30%)

Developmental pathways are likely to differ by epitope

Highly mutated away from their ancestor

CD4bs BNAbs – highly affinity matured

years of infection

Mature BNAb

Unmutated common ancestor

V2/glycan>12 mAbs

V3/glycan>25 mAbs

MPER>5 mAbs

Modified from Burton et al., Science 2012

Long CDRH3 (>25 aa) to penetrate glycan shield

Do long CDR H3s similarly develop gradually?

CAP256

Gray et al 2011, Moore et al 2011

Western Cape

Eastern Cape

Northern Cape

North West

Free State

Northern Province

Gauteng

KwaZuluNatal

Mpumalanga

Durban

Vulindlela

CAP256 targets V2

Twelve related mAbs isolated by B cell culture

Doria-Rose, Schramm, Gorman, Moore et al, Nature, 2014

Weeks post-infection

V1V2-directedNAbs

15 30 38 48 59 119 176* 206

Week Antibody

Somatic mutations (nt)CDR H3 length (aa)

Neutralization (46 strains)

Vl1-51*02 VH3-30*18 BreadthPotency

(IC50, μg/ml)

59 CAP256-VRC26.01 3.9% 8.3% 35 15% 2.93

119

CAP256-VRC26.02 4.9% 8.7% 35 17% 0.40CAP256-VRC26.03 7.4% 8.7% 35 30% 0.06CAP256-VRC26.04 8.1% 9.0% 35 28% 0.25CAP256-VRC26.05 5.4% 10.1% 35 22% 0.10CAP256-VRC26.06 7.4% 10.8% 36 15% 0.16CAP256-VRC26.07 7.7% 11.8% 35 13% 2.02CAP256-VRC26.08 9.8% 11.8% 37 46% 0.14CAP256-VRC26.09 9.8% 14.2% 37 46% 0.08

206CAP256-VRC26.10 3.9% 11.8% 35 24% 0.60CAP256-VRC26.11 13.7% 11.9% 35 24% 0.82CAP256-VRC26.12 8.4% 15.3% 35 7% 0.49

Neutralization breadth starts

CAP256-VRC26 has typical features of anti-V2 broad neutralizing antibodies

HL Kim, A Cupo, R Sanders, IA Wilson, JP Moore, AB Ward

Antibody

BG505SOSIP HIV Env

Ideal for defining the developmental pathways of V2 antibodies with long CDR H3

Jason Gorman and Peter Kwong

PG9 CAP256-VRC26

CAP256-VRC26 heavy chain and light chain antibody sequences

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

VH3-30*18 VL1-52*02

UCA_H UCA_L

Heavy Chain longitudinal

phylogenetic tree

Week38

48

59

119

176

206

Evolutionary distance

0.02

Chaim Schramm and Larry Shapiro

Light Chain longitudinal

phylogenetic tree

The UCA had a 35 amino acid CDR H3 fully formed through VDJ

recombination.

Long CDRH3s

The CAP256-VRC26 unmutated common ancestor had a fully formed long CDR H3

Unmutated common ancestor

Mature BNAb

Long CDRH3s

The CAP256-VRC26 unmutated common ancestor had a fully formed long CDR H3

Was this enough for breadth?

Mature BNAb

Unmutated common ancestor

Development of CAP256-VRC26.01

Nicole Doria-Rose, Ryan Staupe, Nancy Longo, Jinal Bhiman

Weeks post-infection

V1V2-directedNAbs

15 30 38 48 59 119 176* 206

Week Antibody

Somatic mutations (nt)CDR H3 length (aa)

Neutralization (46 strains)

Vl1-51*02 VH3-30*18 BreadthPotency

(IC50, μg/ml)

59 CAP256-VRC26.01 3.9% 8.3% 35 15% 2.93

119

CAP256-VRC26.02 4.9% 8.7% 35 17% 0.40CAP256-VRC26.03 7.4% 8.7% 35 30% 0.06CAP256-VRC26.04 8.1% 9.0% 35 28% 0.25CAP256-VRC26.05 5.4% 10.1% 35 22% 0.10CAP256-VRC26.06 7.4% 10.8% 36 15% 0.16CAP256-VRC26.07 7.7% 11.8% 35 13% 2.02CAP256-VRC26.08 9.8% 11.8% 37 46% 0.14CAP256-VRC26.09 9.8% 14.2% 37 46% 0.08

206CAP256-VRC26.10 3.9% 11.8% 35 24% 0.60CAP256-VRC26.11 13.7% 11.9% 35 24% 0.82CAP256-VRC26.12 8.4% 15.3% 35 7% 0.49

Neutralization breadth starts

Rapid development of neutralization breadth within the CAP256-VRC26 lineage

Chaim Schramm

VRC26-I1VRC26-UCA VRC26-I2 VRC26.01

Rapid development of neutralization breadth within the CAP256-VRC26 lineage

VRC26-I1VRC26-UCA VRC26-I2 VRC26.01

The CAP256-VRC26 UCA, despite having a long CDR H3, can ONLY neutralize the virus that infected CAP256 - no neutralization breadth

CAP256 SI

Rapid development of neutralization breadth within the CAP256-VRC26 lineage

Chaim Schramm

VRC26-I1VRC26-UCA VRC26-I2 VRC26.01

CAP256 SII

Only moderate levels of somatic hypermutation needed for breadth

BreadthBreadth

years of infection ✗Weeks….Unmutated

common ancestor

Mature BNAb with moderate

somatic hypermutation

years of infection

Breadth

UCA

Ontogeny of BNAbs: Looking backwards from breadth

Dramatic viral changes immediately precede CAP256 neutralization breadth

CAP256-VRC26 emerges in peripheral B cell repertoire

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

Dur

ation

of i

nfec

tion

160-171

V1V2 mutations

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

Dramatic viral changes immediately precede CAP256 neutralization breadth

CAP256-VRC26 emerges in peripheral B cell repertoire

Plasma heterologous neutralization develops

160-171

V1V2 mutations

Jinal Bhiman, Daniel Sheward, Molati Nonyane, Bronwen Lambson

Dur

ation

of i

nfec

tion

R166, a crucial part of the CAP256 epitope, undergoes selection pressure as bNAbs

mature

CAP256 infecting

virus

166

94 weeks

p.i.

Emergence of escape mutations drives increased breadth

Jinal Bhiman, Daniel Sheward

166

94 weeks

p.i.

166

All subtype C viruses (n=1,005)

Emergence of new immunotypes drives increased breadth – K166

CAP256 infecting

virus

166

94 weeks

p.i.

166

All subtype C viruses (n=1,005)

Emergence of new immunotypes drives increased breadth – K166

CAP256 infecting

virus

CAP256

CAP256 R166K

Percent breadth

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

Summary

Doria-Rose, Schramm, Gorman, Moore et al, Nature, 2014

V1V2 - Long CDRH3s

CD4bs - highly mutated away from

their ancestor

Which pathway is more amenable to HIV vaccine design?

Requires the engagement of a BCR with a long CDR H3 - these B cells are very rare

Once stimulated, V1V2 BNAbs can develop within months, not years

Immunogens may need to recreate antigenic diversity to drive affinity maturation

Immunogens may need to recreate antigenic diversity to drive affinity maturation

No requirement for long CDR H3, but Ig allele skewing may limit viable BCRs

May need high levels of affinity maturation take years – hard to achieve through vaccination

Acknowledgements Participant CAP256

Slim Abdool Karim, Quarraisha Abdool Karim, Nigel Garrett and the CAPRISA staff

Mascola labNicole Doria-RoseR. Staupe, R. RoarkM. ErnandesRebecca Lynch Rui KongNancy LongoSijy O’DellStephen SchmidtKrisha McKeeMark LouderJohn Mascola

BioinformaticsChaim SchrammZhenhai ZhangCinque SotoIvelin GeorgievLarry Shapiro

Williamson labDaniel ShewardCarolyn Williamson

Kwong labJason GormanMarie PanceraTongqing ZhouBaoshan ZhangIvelin GeorgievYoung Do KwonYongping YangPeter Kwong

NIH Sequencing CoreHolly ColemanBrian SchmidtMorgan ParkJim Mullikin

NIAID NVITALEllen TurkBob Bailer

IAVI NAC, CHAVI-ID and ScrippsAndrew WardHelen KimAlbert CupoJohn MooreRogier SandersWayne KoffIan WilsonDennis Burton

Torrey PinesEmma CrooksJames Binley

U. Texas AustinBrandon DeKoskyGeorge Georgiou

NICDJinal BhimanKurt WibnerMolati NonyaneBronwen LambsonThandeka KhozaMashudu MadzivhandilaLynn Morris