broad and strong immune responses in a trial of a ... · presentation to the aids vaccine 2010...

1

Broad and strong immune responses in a trial of a heterologous DNA prime MVA

boost HIV vaccine among healthy Tanzanian volunteers (HIVIS03)

Presentation to the AIDS Vaccine 2010 Conference, Atlanta, Georgia. 28th Sept. – 1st October 2010

S Aboud1a,2, M Bakari1b, C Nilsson2,3, J Francis4, D Buma5b, C Moshiro1c, EA Aris5b, EF Lyamuya1a, M Janabi5b, K Godoy-Ramirez3, P Earl6, M Robb7, M

Marovich7, N Michael7, B Wahren2,3, K Pallangyo1b, G Biberfeld2,3, F Mhalu1a, E Sandström8, for the HIVIS study group.

1aDepartments of Microbiology and Immunology, 1bInternal Medicine, and 1cEpidemiology and Biostatistics, Muhimbili University of Health and Allied Sciences (MUHAS), Dar es Salaam, Tanzania; 2Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden; 3Swedish Institute for Infectious Disease Control (SMI), Solna, Sweden; 4National Institute for Medical Research (NIMR), Dar es Salaam, Tanzania; 5aDepartments of Internal Medicine and 5bPharmacy, Muhimbili National Hospital (MNH), Dar es Salaam, Tanzania; 6National Institute of Allergy and Infectious Diseases (NIAID)/National Institutes of Health (NIH), Bethesda, Maryland, USA; 7Walter Reed Army Institute for Research (WRAIR), Rockville, Maryland, USA; 8Venhälsan, Karolinska Institutet (KI), Södersjukhuset, Stockholm, Sweden.

2

Objectives

To assess safety and immunogenicity of a HIV-1 plasmid DNA-MVA prime boost vaccine candidate*

To build expertise and capacity in evaluating HIV vaccine candidates in Tanzania

*Previously underwent phase I trial (HIVIS01/02) in Sweden with excellent safety and immunogenicity results. JID 2008; 198:1482-90

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7 plasmid HIV-1 DNA multigene/multiclade vaccine

ORI

CMV

Kanr

poly A

geneInserted

pKCMV

V1 – V5 AV1 – V5 C

R511SA512M

p37 gag Bp37 gag A p24 Gag Ap17 Gag B

RTmut B

D185LD186L

Developed by B Wahren, Dept virology, SMI, Karolinska InstProduced by Vecura

gp160 env Bgp160 env Agp160 env C

rev B

gp120 gp41

LEFT ARM

RIGHT ARM

ORI

CMV

Kanr

poly A

geneInserted

pKCMV


rev B

gp120 gp41


RTmut B

D185LD186L

ORI

CMV

Kanr

poly A

geneInserted

pKCMV


rev B

gp120 gp41

RIGHT ARM


RTmut B

D185LD186L

ORI

CMV

Kanr

poly A

geneInserted

pKCMV


rev B

gp120 gp41

Developed by B Wahren, Dept virology, SMI, Karolinska InstProduced by Vecura


RTmut B

D185LD186L

ORI

CMV

Kanr

poly A

geneInserted

pKCMV


rev B

gp120 gp41

4

MVA* / CMDR boostDeveloped by P Earl and B Moss, Laboratory of Viral Diseases, NIAID, NIHProduced by Walter Reed Army Institute of Research

Deletion IIIDeletion II

MVAgag protease / RTgp150 env

Subtype ECM235

Subtype ACM240

mH5 mH5

*Modified Vaccinia Ankara



Subtype ECM235

Subtype ACM240

mH5 mH5


Developed by P Earl and B Moss, Laboratory of Viral Diseases, NIAID, NIHProduced by Walter Reed Army Institute of Research



Subtype ECM235

Subtype ACM240

mH5 mH5


Developed by P Earl and B Moss, Laboratory of Viral Diseases, NIAID, NIHProduced by Walter Reed Army Institute of Research



Subtype ECM235

Subtype ACM240

mH5 mH5


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Study DesignRandomized, Double Blind, Placebo controlledArm Number DNA immunization MVA boost

I 20 DNA 3.8mg IM by Biojector MVA 108 pfu IM

II 20 DNA 1 mg ID by Biojector MVA 108 pfu IM

IIIa 10 Saline IM by Biojector Saline IM

IIIb 10 Saline ID by Biojector Saline IM

1 2 3 4 5 6 7 8 90 10

HIV-MVA boost with needleHIV-DNA prime with Bioject

21 months 1 2 3 4 5 6 7 8 90 10

HIV-MVA boost with needleHIV-DNA prime with Bioject

21 months

Police Officer Cohort in Dar es Salaam

Inclusion Criteria• Voluntary Informed Consent• Age >18 and <40 years• HIV negative by Ag-Ab ELISA• “Healthy” by Clinical and Laboratory Evaluation• At low-risk for acquiring HIV infection

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Methods• IFN- γ ELISpot responses were measured on

fresh cells (within 6 hours of collection) after stimulation with HIV-1 peptide pools– The criteria for positive ELISpot responses were

>55 spots/106 PBMCs and 4 times the medium background and the baseline value.

• 4-colour intracellular cytokine staining (ICS) Gag-specific IFN-gamma/IL-2 production by CD4 and CD8 T-cells– CD4>3 SI and >0.05%; CD8 >3 SI and >0.1%

• Lymphoproliferation assay (LPA) by 3H-thymidine uptake using AT-2 treated antigens kindly provided by Jeff Lifson

88

Peptide pools used in ELISpot

All peptides have 15-mers with 10 amino acid (aa) overlap except * the WR peptide pools which have peptides with 11 aa overlap. †DNA vaccine clade A and B specific peptides.

Peptide pool ID Protein Peptide number Subtype

Gag I† p17 1-26 B

Gag II† p24 27-71 A

Env I† gp120, including V1

and V2

1-50 A/B

Env II† gp120, including V3-V5

51-100 A

Env III† gp41 101-169 B

Gag WR* p6, p7, p17, p24 1-160 A

Env WR* Env 1-177 E

IFN-γ ELISpot Responses After HIV-DNA priming and MVA boosting

Responders 2 weeks after the 1st HIV-MVA boost

Responders 2-4 weeks after the 2nd HIV-MVA boost

Peptidepool

% n Peptidepool

% n

Gag or Env

100 35/35 Gag or Env

97 28/29

Gag 100 35/35 Gag 93 27/29Env 89 31/35 Env 79 23/29

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Gag

I

Gag

II

Gag

WR

Env

I

Env

II

Env

III

Env

WR

1

10

100

1000

10000

SFC

/mill

ion

PB

MC

s

Gag

I

Gag

II

Gag

WR

Env

I

Env

II

Env

III

Env

WR

1

10

100

1000

10000

SFC

/mill

ion

PB

MC

s

Magnitude of IFN-γ ELISpot responses after the first and second HIV-MVA boost

2 weeks after the 1st HIV-MVA, all 35 (100%) vaccinees responded

Peptide pool Peptide pool

Gag

I

Gag

II

Gag

WR

Env

I

Env

II

Env

III

Env

WR

1

10

100

1000

10000

SFC

/mill

ion

PB

MC

s

Gag

I

Gag

II

Gag

WR

Env

I

Env

II

Env

III

Env

WR

1

10

100

1000

10000

SFC

/mill

ion

PB

MC

s


2 to 4 weeks after the 2nd HIV-MVA, 28/29 (97%) vaccinees responded

2 weeks after the 1st HIV-MVA, all 35 (100%) vaccinees responded

Gag

I

Gag

II

Gag

WR

Env

I

Env

II

Env

III

Env

WR

1

10

100

1000

10000

SFC

/mill

ion

PB

MC

s

Gag

I

Gag

II

Gag

WR

Env

I

Env

II

Env

III

Env

WR

1

10

100

1000

10000

SFC

/mill

ion

PB

MC

s


Higher magnitude of IFN-γ ELISpotresponses to Env in i.d. vs. i.m. HIV-

DNA-primed vaccinees

* *

* Significantly higher responses with i.d. than i.m. prime

HIV-specific ICS responses 4 weeks after the second HIV-MVA boost

Gag-specific IFN- γ/IL-2 production by CD4 and CD8 T-cells

RespondersT-cells % nCD4+ 55 16/29CD8+ 59 17/29CD4+ or CD8+ 86 25/29

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MN KNH TZA CM SUPT Jurkat0.1

1

10

100

1000

10000

Stim

ulat

ion

inde

x (S

I)


1

10

100

1000

10000

Stim

ulat

ion

inde

x (S

I)Cross-clade LPA responses 2 weeks and

6 months after the 2nd HIV-MVA boost2 weeks after the 2nd HIV-MVA, all of the 25 vaccinees (100%) showed positive LPA responses to HIV-1antigens of clades B, A and A_E

Subtype B A C A_ESubtype B A C A_E


1

10

100

1000

10000

Stim

ulat

ion

inde

x (S

I)


1

10

100

1000

10000

Stim

ulat

ion

inde

x (S

I)

Subtype B A C A_E

2 weeks after the 2nd HIV-MVA, all of the 25 vaccinees (100%) showed positive LPA responses to HIV-1antigens of clades B, A and A_E


1

10

100

1000

10000

Stim

ulat

ion

inde

x (S

I)


1

10

100

1000

10000

Stim

ulat

ion

inde

x (S

I)

Subtype B A C A_ESubtype B A C A_EB ASubtype B A C A_EB ASubtype B A C A_EB ASubtype A C A_EA Subtype B A C A_ESubtype B A C A_EB ASubtype B A C A_EB ASubtype B A C A_EB ASubtype A C A_EA


1

10

100

1000

10000

Stim

ulat

ion

inde

x (S

I)


1

10

100

1000

10000

Stim

ulat

ion

inde

x (S

I)

Subtype B A C A_ESubtype B A C A_EB ASubtype B A C A_EB ASubtype B A C A_EB ASubtype A C A_EA

6 months after the 2nd HIV-MVA, 16 of the 18 vaccinees (89%) had positive LPA responses

2 weeks after the 2nd HIV-MVA, all of the 25 vaccinees (100%) showed positive LPA responses to HIV-1antigens of clades B, A and A_E


1

10

100

1000

10000

Stim

ulat

ion

inde

x (S

I)


1

10

100

1000

10000

Stim

ulat

ion

inde

x (S

I)

Subtype B A C A_ESubtype B A C A_EB ASubtype B A C A_EB ASubtype B A C A_EB ASubtype A C A_EA

Antibody responses after the 1st and 2nd HIV-MVA boost

Test 1st HIV-MVA 2nd HIV-MVAGp 160 ELISA

7/33 (21%) 26/29 (90%)

Abbott MurexELISA

0/35 (0%) 30/30 (100%)

EnzygnostPlus ELISA

0/35 (0%) 30/30 (100%)

Inno-Liaimmunoblot

0/35 (0%) 30/30 (100%)14

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Conclusions• This heterologous HIV DNA prime MVA

boost vaccine approach in Tanzanian volunteers demonstrated:– Broad and high immune responses– Both CD4+ and CD8+ T-cell responses– Good durability of immune responses 6

months after the second HIV-MVA boost– 100% binding antibody responses after the

second HIV-MVA boost • Capacity built through HIVIS03 paved the

way for EDCTP-funded TaMoVaC Project aimed at optimizing DNA vaccine delivery

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Acknowledgements

• Police volunteers• EU and Sida (Sweden)• Swedish Embassy, Tanzania• EDCTP• WHO-UNAIDS and AAVP• Investigators, collaborators and support staff in:

–Tanzania; at MUHAS, MNH, Tanzania Police Force–Sweden; at Karolinska Institute, Swedish Institute for Infectious Disease Control, Southern Hospital–United States of America; at WRAIR and LVD/NIH

• Tanzania Government, in particular the MoH & SW through NACP


After first HIV-MVA• Mean: 784p<0.005 (Wilcoxon test)• SD: 591• Min-Max: 70-2285• Median: 625• 25th percentile: 250p<0.02 (paired T-test)

After second HIV-MVA• 452

• 388• 10-1560• 335• 150

After first HIV-MVA• Mean: 784p<0.005 (Wilcoxon test)• SD: 591• Min-Max: 70-2285• Median: 625• 25th percentile: 250p<0.02 (paired T-test)

After second HIV-MVA• 452

• 388• 10-1560• 335• 150

Gag

I

Gag

II

Gag

WR

Env

I

Env

II

Env

III

Env

WR

1

10

100

1000

10000

SFC

/milli

on P

BM

Cs

Gag

I

Gag

II

Gag

WR

Env

I

Env

II

Env

III

Env

WR

1

10

100

1000

10000

SFC

/milli

on P

BM

Cs



6 months after the 2nd HIV-MVA, 17/26 (65%) vaccinees responded

Gag

I

Gag

II

Gag

WR

Env

I

Env

II

Env

III

Env

WR

1

10

100

1000

10000

SFC

/milli

on P

BM

Cs

Gag

I

Gag

II

Gag

WR

Env

I

Env

II

Env

III

Env

WR

1

10

100

1000

10000

SFC

/milli

on P

BM

Cs


6 months after the 2nd HIV-MVA, 17/26 (65%) vaccinees responded

Gag

I

Gag

II

Gag

WR

Env

I

Env

II

Env

III

Env

WR

1

10

100

1000

10000

SFC

/milli

on P

BM

Cs

Gag

I

Gag

II

Gag

WR

Env

I

Env

II

Env

III

Env

WR

1

10

100

1000

10000

SFC

/milli

on P

BM

Cs



1

10

100

1000

10000

Sti

mu

lati

on

ind

ex (

SI)


1

10

100

1000

10000

Sti

mu

lati

on

ind

ex (

SI)

Strong cross-clade LPA responses 2 weeks after the 1st and 2nd HIV-MVA

boost2 weeks after the 1st HIV-MVA, all 32 vaccinees had positive LPA responses

2 weeks after the 2nd HIV-MVA, all of the 25 vaccinees (100%) had positive LPA responses

Subtype B A C A_E Subtype B A C A_ESubtype B A C A_E Subtype B A C A_E


1

10

100

1000

10000

Sti

mu

lati

on

ind

ex (

SI)


1

10

100

1000

10000

Sti

mu

lati

on

ind

ex (

SI)

Subtype B A C A_E Subtype B A C A_E

2 weeks after the 1st HIV-MVA, all 32 vaccinees had positive LPA responses


1

10

100

1000

10000

Sti

mu

lati

on

ind

ex (

SI)


1

10

100

1000

10000

Sti

mu

lati

on

ind

ex (

SI)


2 weeks after the 2nd HIV-MVA, all of the 25 vaccinees (100%) had positive LPA responses

2 weeks after the 1st HIV-MVA, all 32 vaccinees had positive LPA responses


1

10

100

1000

10000

Sti

mu

lati

on

ind

ex (

SI)


1

10

100

1000

10000

Sti

mu

lati

on

ind

ex (

SI)


Binding antibodies after the 1st and 2nd HIV-MVA boost

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<100

100

1000

10000gp

160

ELIS

A tit

er

2 months after the 1st

HIV-MVA vaccination

1 month after the 2nd

HIV-MVA vaccination

HIVIS03 Summary

Immune responses 2-4 weeks after the 2nd HIV-MVA boostIFN-γ ELISpot 28/29 (97%)ICS Gag-specific IFN- γ/IL-2 production by T-cells:CD4+ CD8+CD4+ or CD8+

16/29 (55%)17/29 (59%)25/29 (86%)

LPA responses 25/25 (100%)Binding antibodies to gp160 26/29 (90%)

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