shaping ctl immunodominance with abstract #320 conserved

*MembersoftheBCN01studygroup§  IrsiCaixa-HIVACAT,FundacióLluitacontralasida,HospitalGermansTriasiPujol,

Badalona,Spain:BeatrizMothe,PepColl,PatriciaCobarsi,RoserEscrig,NuriaPérez-Alvarez,AlvaroSanchez-Bernabeu,MiriamRosàs,M.CarmenPuertas,SaraMorón,

JavierMartinez-Picado,BonaventuraClotet,ChristianBrander.§  HospitalClínic-HIVACAT,IDIBAPS,Barcelona,Spain:ChristianManzardo,CristinaRovira,

IreneRuiz,J.MariaGatell,J.MariaMiró.

§  BCN-Checkpoint,ProjectedelsNoms,Barcelona,Spain:PepColl,JorgeSaz,MichaelMeulbroek,FerranPujol§  NuffieldDept.Medicine(NDM)&TheJennerInstitute,UniversityofOxford,Oxford,

UK:LucyDorrell,NicolaBorthwick.AlisonCrook,TomášHanke.FundingThisstudyhasbeenpartiallyfundedbyHIVACATprogramandFundacióGloriaSoler.BMisaJoanRodésinvestigatorfromtheISCIII(JR13/00024),Madrid,Spain.

Painsc

ale

Life-threatening

Severe

Moderate

Low

Nosymptoms

ChAdV63 MVA

B.Mothe1,2,3,C.Manzardo4,P.Coll1,2,SaraMorón1,L.Dorrell5,B.Clotet1,2,3,J.Martinez-Picado1,3,6,T.Hanke7andC.Brander1,3,6fortheBCN01studygroup*

1IrsiCaixaAIDSResearchInstitute-HIVACAT,Badalona,Spain;2FundacióLluitacontralaSida,Badalona,Spain;3UniversitatdeVic-CentraldeCatalunya,Uvic-UCC,Spain;4HospitalClinic-IDIBAPS,Barcelona,Spain;5NuffieldDept.Medicine(NDM),UniversityofOxford,UK;6ICREA,Catalunya,Spain;7TheJennerInstitute,Oxford,UK

ShapingCTLImmunodominanceWithConservedHIVVaccinesAfterEarlyTreatment(BCN01)

BackgroundTherapeutic T-cell vaccines targeting the most conserved regions of the HIV-1 proteome have the potential toenhancehostimmunecontrolandfacilitateclearanceofthelatentreservoir.HIVconsvvaccines(Fig1)vectoredbychimpanzeeadenovirus(ChAdV63)andmodifiedvacciniavirusAnkara(MVA)havebeenshowntoinducehighlevelsofeffectorTcellsinhealthyindividuals(HIVCORE02trial;Borthwick,2014).

Fig2.FlowChartofStudyDesign

BCN01 (NCT01712425) is a phase I, multicenter trial toevaluatethesafety, immunogenicityandimpactonthelatentreservoirof acombinedChAdV63-MVA.HIVconsvvaccine in early-treated individuals (<6m from HIV-1infection, n=24) who initiated TDF/FTC/RAL 1 wk afterdiagnosis.(Table1)

2.Localandsystemiceventsaftervaccinationoccurredin 22/24 individuals, mostly severity grade 1-2 andtransiently (48 hours). Local pain was more oftenreportedwithMVAthanChAdV63vaccination.

Fig1.SchematicrepresentationofHIVconsvImmunogenRepresentationofthehumanizedgenecodingfortheHIVconsvimmunogen,whichcodesforthe14mostconservedregionsoftheconsensusHIV-1Gag,Pol,Vif,andEnvproteins.Capitallettersabovebarsindicatethecladeusedforthederivationoftheconsensusaasequenceforeachconservedsegment.Totallenghtoftheinsert778aaHIVconsvimmunogenisvectoredbychimpanzeeadenovirus63,ChAdV63.HIVconsv(C)andmodifiedvacciniavirusAnkara,MVA.HIVconsv(M).AdaptedfromLetourneau,2007

Immunogenicity to theHIVconsvvaccine insertandtherest of the HIV-1 proteome was assessed by IFNgELISPOT in cryopreserved PBMC. Proviral DNA wasquantifiedinpurifiedCD4+T-cellsbydropletdigitalPCR.Single-copy assay was performed to investigatepotentialviralreactivationduringvaccinations.

Individuals received ChAdV63.HIVconsv (5x1010vp, im)and a MVA.HIVconsv boost (2x108pfu, im) 8 or 24 wkafterwards(ShortvsLongregimen)andwerefollowedfor6months. 24unvaccinatedcontrolswere includedtocompareviralreservoirdecayduring1styearofearly-treatmentinitiation.(Fig2.FlowChart)

Tabe1.PopulationCharacteristics.

ShortArm(0-8w;n=12)

LongArm(0-24w;n=12)

UnVax(Controls;n=24)

MedianAgeatBSL(range) 38(27-48) 41(30-54) 34(19-62)

Sexratio(men/female) 12/0 11/1 12/0

HTS/HMS 12/0 9/3 12/0

MediandayssinceHIVtransmission(&range)tocARTinitiation

91(51-120) 97(65-206) 70(15-101)

V0HIV-1Dx

V2(4w)

V3(12w)

V4(24w)

ChAdprime MVAboost

V8(48w)

V12(+24)

V1(0w)

24 week

V11(+12)

VACCINE Long Arm (n=12)

V0HIV-1Dx

V2(4w)

V3(12w)

V4(24w)

ChAdprime

V7(32w)

MVAboost

V11(+24)

V1(0w)

8 week

VACCINE Short Arm(n=12)

V10(+12)

UNVACCINATED CONTROLS (n=24)

V2(4w)

V3(12w)

V4(24w)

V5(36w)

V6(48w)

V8(72w)

V1(0w)

V0HIV-1Dx

V7(60w)

cART(TDF/FTC+RAL)iniNaNon(within1weeka@erdiagnosis)infecNon

confirmed<6months

confirmed<6months

confirmed<6months

0 4 12 24 36 48 600

100200300400500600700800900

0

1

2

3

4

5

6

Short Arm Short ArmLong Arm Long ArmControls Controls

Viral load CD4 count and Ratio CD4/CD8

ChAdMVAshort MVAlong

Time (weeks)

Med

ian

CD

4 T

cell

coun

t (c

ells

/ul) M

edian log10 (viral load, cp/m

l)

0 4 12 24 36 48 60

0.5

1.0

1.5

2.0

Time (weeks)

Med

ian

ratio

CD

4/C

D8


0 4 12 24 36 48 600

100200300400500600700800900

0

1

2

3

4

5

6




Time (weeks)

Med

ian

CD

4 T

cell

coun

t (c

ells

/ul) M


l)

0 4 12 24 36 48 60

0.5

1.0

1.5

2.0

Time (weeks)

Med

ian

ratio

CD

4/C

D8


0 4 12 24 36 48 600

100200300400500600700800900

0

1

2

3

4

5

6




Time (weeks)

Med

ian

CD

4 T

cell

coun

t (c

ells

/ul) M


l)

0 4 12 24 36 48 60

0.5

1.0

1.5

2.0

Time (weeks)

Med

ian

ratio

CD

4/C

D8


CD4countorRatioCD4/CD8

0 4 12 24 36 48 600

100200300400500600700800900

0

1

2

3

4

5

6




Time (weeks)

Med

ian

CD

4 T

cell

coun

t (c

ells

/ul) M


l)

0 4 12 24 36 48 60

0.5

1.0

1.5

2.0

Time (weeks)

Med

ian

ratio

CD

4/C

D8


1. Patients Characteristics, Immune Recovery and Viralsuppression dynamics are consistent with an earlytreatedHIV+population(Integrase-InhibitorbasedART)

ViralLoad(pVL)

Heterologous prime/boost vaccination with ChAdV63 andMVA.HIVconsv was a safe strategy to induce new and/or shift pre-existing immune response towards conserved regions of HIV-1 in acohortofearly-treatedindividuals.Reservoirdecayduringfirstyearofearly-ARTwasnotfurtherimpactedbyHIVconsvvaccinations.

ConclusionsandFuturework

This is the first therapeutic vaccine trial able to demonstrate arefocusingoftheCTLimmunodominancepatternstowardsconservedregions of HIV-1 andmay provide the base for effective kick and killstrategies(Roll-overstudyBCN02-Romi,NCT02616874,enrolling2016)

Methods Results

3. HIVconsv induced responses over vaccination. 22 individuals (92%) mounted de-novo HIVconsv-specific Tcellresponses during vaccination (not detectable before cART initiation). Responses were increased in 50% ofparticipantsafterChAdV63primeandin100%ofparticipantsafterMVAboostervaccination.

ShortArm LongArm

0%#

10%#

20%#

30%#

40%#

50%#

60%#

70%#

80%#

90%#

100%#

P1# P2# P3# P4# P5# P6#

Percentage)of)responders)by)HIVconsv)regions)

Responders#pre6vax# responders#at#peak#

TotalMagnitudeofHIVconsv-specificIFN-gpositiveTcellsovervaccinationperiodbyvaccinationstudygroups

PercentatgeofvaccinatedindividualswithanincreaseintotalHIVconsv-specificreponsesbeforeandaftervaccination(left)andbypeptidepools(right).

ChangeintotalMagnitudeofHIVconsv-specificTcellsbeforeandaftervaccination

0%#

10%#

20%#

30%#

40%#

50%#

60%#

70%#

80%#

90%#

100%#

BSL$ ChAd$ MVA$ 24w$a/er$last$vax$

Percentage)of)responders)

5.ChangeindominancepatternsofHIVresponses.NosignificantexpansionofT-cellstargetingHIV-1regionsoutsidethevaccineinsertwasnoted,reflectiveofaneffectiveshiftofCTLimmunitytowardsconservedregions(58%)

6.Noreactivationobservedduringvaccination.

0

250

500

750

1000

1250

0

25

50

75

100

MagnitudeofHIVconsvspecificresponses(Median,IQR)FocusofHIVconsvovertotalHIV-specificresponses(Median)

BSL C0 C1wk C4wk M0 M1wk M4wk M24wk

HIVco

nsvIFNγSF

C/10

⁶ PBM

C Focus(%)HIVconsvover

totalHIV-specificresponses

0

250

500

750

1000

1250

0

25

50

75

100

MagnitudeofHIVconsvspecificresponses(Median,IQR)FocusofHIVconsvovertotalHIV-specificresponses(Median)

BSL C0 C1wk C4wk M0 M1wk M4wk M24wk

HIVco

nsvIFNγSF

C/10

⁶ PBM

C Focus(%)HIVconsvover

totalHIV-specificresponses

ChAd MVA ChAd MVA

ShortArm LongArm

MagnitudeandFocus(median)ofHIVconsv-specificIFN-gpositiveTcellsovertotalHIV-specificresponsesovervaccinationperiodbyvaccinationgroups

FocusofHIVconsv-specificresponsesinallindividualsbeforecART,beforevaccination,atpeakofinducedresponsesand

24weeksafterboostinallindividuals

BOX for pools for BCN01 (ChAd-MVA.HIVconsv trial) IMMUNOGENICITY

1 2 3 4 5 6A HIVconsv HIVconsv HIVconsv HIVconsv HIVconsv HIVconsv

p1 p2 p3 p4 p5 p6

B HIVconsv HIVconsv HIVconsv HIVconsv HIVconsv HIVconsv

p1 p2 p3 p4 p5 p6

COUT HIV-1OUT HIV-1OUT HIV-1OUT HIV-1OUT HIV-1OUT HIV-1

BCN01-OUT Gag-1

BCN01-OUT Gag-2

BCN01-OUT Pol-1

BCN01-OUT Pol-2

BCN01-OUT Pol-3

BCN01-OUT V-T

DOUT HIV-1OUT HIV-1OUT HIV-1OUT HIV-1OUT HIV-1OUT HIV-1

BCN01-OUT Gag-1

BCN01-OUT Gag-2

BCN01-OUT Pol-1

BCN01-OUT Pol-2

BCN01-OUT Pol-3

BCN01-OUT V-T

EOUT HIV-1OUT HIV-1OUT HIV-1OUT HIV-1OUT HIV-1OUT HIV-1

BCN01-OUT Env-1

BCN01-OUT Env-2

BCN01-OUT Env-3

BCN01-OUT Env-4

BCN01-OUT Nef

BCN01-OUT Acc

FOUT HIV-1OUT HIV-1OUT HIV-1OUT HIV-1OUT HIV-1OUT HIV-1

BCN01-OUT Env-1

BCN01-OUT Env-2

BCN01-OUT Env-3

BCN01-OUT Env-4

BCN01-OUT Nef

BCN01-OUT Acc

G HTI HTI HTI HTI HTI Mouse/Mac

Gag-p1 Gag-p2 Pol-p1 Pol-p2 Vif-Nef 1

H

NEG NEG NEG NEG PHA CEF

FOCU

S:

HIVco

nsv/To

tal=

HIVco

nsv/(H

IVco

nsv+OUT)

LayoutofIFNgELISPOTplatesforImmunogenicitystudiesassessingHIVconsvspecificresponsesaswellasresponsesto

HIVregionsnotincludedintheimmunogen.Overlappingpeptides(15mers,11)wereusedinpools.

ShortArm LongArm

M0 M1

0

10

20

30

LOD

copi

es H

IV-1

RN

A/m

l pla

sma

M0 M1

0

10

20

30

LOD

copi

es H

IV-1

RN

A/m

l pla

sma

M0 M1

0

10

20

30

LOD

copi

es H

IV-1

RN

A/m

l pla

sma

M0 M1

0

10

20

30

LOD

copi

es H

IV-1

RN

A/m

l pla

sma

Quantificationofresidualplasmaviremiabeforeand1weekafterMVA.HIVconsvvaccination(peakofimmunogenity)inbotharmsbyRT-PCR.

7.Reservoirdecay(proviralDNA)wasequalinallgroupsShort prime/boost regimen

24 58

0

2000

4000

6000

8000

10000

p=0.0010

Time after cART initiation (weeks)To

tal H

IV-1

DN

A c

op

ies

/10

6 C

D4

+ T

ce

lls

Controls

24 60

0

2000

4000

6000

8000

10000 p<0.0001


tal H

IV-1

DN

A c

op

ies

/10

6 C

D4

+ T

ce

lls

Long prime/boost regimen

24 62

0

2000

4000

6000

8000

10000

p=0.0005


tal H

IV-1

DN

A c

op

ies

/10

6 C

D4

+ T

ce

lls

Short prime/boost regimen

24 58

0

2000

4000

6000

8000

10000

p=0.0010


tal H

IV-1

DN

A c

op

ies

/10

6 C

D4

+ T

ce

lls

Controls

24 60

0

2000

4000

6000

8000

10000 p<0.0001


tal H

IV-1

DN

A c

op

ies

/10

6 C

D4

+ T

ce

lls


24 62

0

2000

4000

6000

8000

10000

p=0.0005


tal H

IV-1

DN

A c

op

ies

/10

6 C

D4

+ T

ce

lls

Short prime/boost regimen

24 58

0

2000

4000

6000

8000

10000

p=0.0010


tal H

IV-1

DN

A c

op

ies

/10

6 C

D4

+ T

ce

lls

Controls

24 60

0

2000

4000

6000

8000

10000 p<0.0001


tal H

IV-1

DN

A c

op

ies

/10

6 C

D4

+ T

ce

lls


24 62

0

2000

4000

6000

8000

10000

p=0.0005


tal H

IV-1

DN

A c

op

ies

/10

6 C

D4

+ T

ce

llsShortArm LongArm Controls

p=0.0010 p=0.0005 p<0.0001

CopiesoftotalHIVDNApermillionCD4TcellsmeasuredbydropletdigitalPCRforeachstudygroup(WilcoxonsignedrankTtest)

Decay

Short Long Controls

-1

0

1

2

3 p=0.9370

Rat

io [I

ncre

ase w

k60-

wk2

4/To

tal H

IV-1

DN

Aw

k24]

p=0.94

RaMo

DecayoftotalHIVDNA(w24àw60afterearly_cARTinitiationinvaccinatedandnon-vaccinatedindividuals

Decay(ΔReswk60-24/Reswk24)

Abstract#[email protected]

[email protected]

shaping ctl immunodominance with abstract #320 conserved

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