*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
ChAdMVAshort MVAlong
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
ChAdMVAshort MVAlong
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
ChAdMVAshort MVAlong
CD4countorRatioCD4/CD8
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
ChAdMVAshort MVAlong
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
Time after cART initiation (weeks)To
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
Time after cART initiation (weeks)To
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
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
Time after cART initiation (weeks)To
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
Time after cART initiation (weeks)To
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
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
Time after cART initiation (weeks)To
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
Time after cART initiation (weeks)To
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]
