ana guimar ã es- walker,

Ana Guimarães- Walker, I. Cebere, N. Mackie, T. Hanke, P. Fast, L. Dally, J. Weber and A. J. McMichael.

Safe administration of DNA (pTHr.HIVA) and

MVA.HIVA to 169 HIV-1 uninfected

volunteers enrolled in Phase I/II trials in the UK.

XVI International AIDS ConferenceToronto, Canada.

2MRC Human Immunology Unit - WIMM - Oxford University

Vaccine CandidatesDNA vaccine (pTHr.HIVA)

Plasmid

Modified Vaccinia Ankara (MVA.HIVA)

Non-replicant live attenuated vector

HIVA-AClade A HIV-1 gag p17 and p24

25 CTL epitopes linked end-to-end (env, gag, pol and nef)

Humoral and cellular responses in rodents and macaques.

Hanke T, Nat Med 2000Hanke T, Nat Med 2000

SIV-specific CTL response in rhesus monkeys.

Wee EG, J Gen Virol 2002Wee EG, J Gen Virol 2002

Multifunctional HIV-1 specific T-cell responses in healthy seronegative subjects.

Goonetilleke N, J Virol, May 2006Goonetilleke N, J Virol, May 2006 Specific T-cell responses in

HIV-1 infected subjects.Dorrell L., J Virol, May 2006Dorrell L., J Virol, May 2006

3MRC Human Immunology Unit - WIMM - Oxford University

Study DesignTime Time pointspoints (WK)(WK)

00 33 44 88 1212 2020 2424 5252 104104DNA0.1 mg001

n=18

003n=8

005(n=9)

016n=18

006n=18

Early boost

DNA0.5 mg

DNA0.1 mg

DNA0.5 mg

DNA0.5 mg

DNA4 mg

DNA0.5 mg

DNA2 mg

DNA2 mg

DNA4 mg

MVA5x107 pfu

MVA5x107 pfu

MVA5x107 pfu

MVA5x107 pfu

MVA5x107 pfu

MVA5x107 pfu

MVA5x107 pfu

MVA5x107 pfu

MVA1x108 pfu

MVA1x108 pfu

MVA1x108 pfu

Late boost

END

END

END

END

END

END

END

4MRC Human Immunology Unit - WIMM - Oxford University

DemographicsAge #001 #003 #005 #006 #016

Median 34 34 43 36 43

Range 20-57 22-54 21-58 18-59 20-56

Gender

57% 43%

Female Male

Ethnicity 91%

4%3%

2%

Asian Black Caucasian Other

(n=73)

(n=96)

(n=155)

(n=6)(n=5

)

(n=3)

5MRC Human Immunology Unit - WIMM - Oxford University

Clinical Definitions Adverse Events:

Vaccine reactions Other AEs Solicited events Unsolicited events “Reactogenicity” “Ongoing

reactogenicity”

Types of cutaneous reactions:

• Pain, tenderness, itchiness• Erythema• Swelling x induration• Blister, vesicle ulceration• Rash

Grading parameters:• % of Arm circumference• Size• Duration• Use of medication• Other

Grading criteria cutaneous reactions:

6MRC Human Immunology Unit - WIMM - Oxford University

Maximum Local Reactions

(per volunteer during the 28 days post vaccination)

Severity

0%87%

13%

Mild Moderate Severe

volunteers x local reactions

18 18

8

3 4

11

04

16

1 0

16

0 0

16

0 0

8

0 03

0

5

10

15

20

Placebo DNA MVA

None Pain Tenderness Erythema

Induration Skin damage Crust/ scab

n=8 n=8 n= 16

# Trial 016 (n=24)

Typical local reactions:• Following DNA.HIVA Following MVA.HIVA

Pain/tenderness Pain/tendernessErythemaInduration

N vol

7MRC Human Immunology Unit - WIMM - Oxford University

Maximum Systemic Reactions

(per volunteer during the 28 days post vaccination)

Severity

0%84%

16%

Mild Moderate Severe

volunteers x systemic reactions

5 6 51 0

6

1 1

8

1 032 2

7

0 0 11 03

0 0 10

5

10

15

20

Placebo DNA MVA

None Chill Malaise Myalgia

Headache Fever Nausea Vomiting

# Trial 016 (n=24)

Typical local reactions:• Following DNA.HIVA Following MVA.HIVA

None Malaise/ myalgiaHeadache

N vol

n=8 n=8 n= 16

8MRC Human Immunology Unit - WIMM - Oxford University

0

20

40

60

80

100

Placebo(N=63)

Vaccine(N=190)

Placebo(N=42)

Vaccine(N=377)

None Unlikely Possible Probable

0

20

40

60

80

100

Placebo(N=63)

Vaccine(N=190)

Placebo(N=42)

Vaccine(N=377)

Mild Moderate Severe

Non-Serious Adverse Events (Excludes Local and Systemic Vaccine

Reactions)Severity Relationship

DNA DNA

MVAMVA

%

%

Over 1000 AEs reported across trials

~ 30% of AEs considered possibly or probably related to vaccine

< 10% of events related to vaccine graded as moderate or severe

9MRC Human Immunology Unit - WIMM - Oxford University

Serious Adverse Events (SAEs)

SAEs reported for 6 volunteers:

1.1. Hospitalization: fever, malaise, nausea & Hospitalization: fever, malaise, nausea & headacheheadache 2.2. Removal of gallbladderRemoval of gallbladder

Umbilical hernia repairUmbilical hernia repair

3.3. Hospitalization: cycle accidentHospitalization: cycle accident

4.4. Excavation of sinusesExcavation of sinusesRemoval of nasal polypsRemoval of nasal polypsRealignment of nasal septumRealignment of nasal septum

5.5. MesotheliomaMesothelioma

6.6. Hysterectomy and bilateral oophorectomyHysterectomy and bilateral oophorectomy

No SAES caused by Vaccine.

Days (n) Enrolment

Days (n) Last vac

04

30 02

787878

313-44830

179

175 91

03

50

261-12630

04

5050

10MRC Human Immunology Unit - WIMM - Oxford University

Laboratory assessment No positive ELISA HIV results.

No evidence of induction of auto-immune antibodies.

Liver function tests Grade 3 or greater laboratory values

leading to discontinuation of vaccinations (n=02 volunteers).

11MRC Human Immunology Unit - WIMM - Oxford University

Immunogenicity of DNA.HIVA and MVA.HIVA

Positive Responders across assays in 016 Trial

Subject Cultured

ELISPOT

3H Thymidine Proliferation

Ex vivo ELISPOT

ELISPOT +IL-7/IL-15

CFSE PROLIF.

IFN- WBICS

DNA-MVA -

1 77692 224.21 959 2165 0.56 0.46

2 104313 11.62 103 166 0.07 0.01

3 32062 44.41 514 ND4 ND 0.02

4 41812 7.33 36 86 0.03 0.01

5 14632 8.50 31 128 0.01 0.03

6 3882 4.28 30 96 0.00 0.02

7 4132 0.93 4 18 0.01 0.01

8 17692 1.59 5 13 0.01 0.01

MVA

9 2494 0.36 28 165 (CTL) ND 0.01

10 1250 (CTL) 0.15 -1 -3 ND ND

11 356 (CTL) 0.97 0 15 ND 0.00

12 3838 0.55 6 ND ND 0.00

13 ND 1.01 0 16 ND 0.00

14 ND 0.30 3 -18 ND 0.00

15 ND 0.69 -1 31 ND 0.00

16 ND 0.17 11 16 ND 0.00

PLACEBO

n=8 No positive responses

1 Background subtracted responses to Gag (CTL string response indicated in brackets). Responses shown are from Wk9 in the DNA/MVA arm and Wk1 in the MVA only arm2 Response from Wk10 (2 weeks post MVA) 3 Response from W12 (4 weeks post MVA)4 ND= no data available for Pool 90 specific responses

Responders in ELISPOT assays

0

4

0 0

21

0

4

8

0

10

DNA-MVA MVA Placebo

Cultured Ex-vivo +Il-7/ Il-15n=8 n=8

n=8

12MRC Human Immunology Unit - WIMM - Oxford University

Conclusions – Safety Overall both DNA.HIVA and MVA.HIVA vaccines were

safe and well tolerated.

Most local or systemic reactions following vaccination with DNA or MVA vaccinations were mild.

Local and systemic reactions were more frequently observed following MVA vaccination, compared to DNA-HIVA vaccination.

Ensuring consistency in the grading criteria is crucial within a programme consisting of overlapping trials.

13MRC Human Immunology Unit - WIMM - Oxford University

Acknowledgments: MRC Human MRC Human Immunology OxfordImmunology OxfordProfessor Andrew Professor Andrew

McMichaelMcMichaelDr Inese CebereDr Inese Cebere

Dr Jo RobertsDr Jo RobertsDr Nilu GoonetillekeDr Nilu Goonetilleke Dr Susana PinheiroDr Susana PinheiroDr Thomas HankeDr Thomas HankeMs Althea ThomasMs Althea Thomas Ms Denise BrownMs Denise Brown

Ms Nicola WinstoneMs Nicola WinstoneMr Stephen MooreMr Stephen MooreMs Vanessa LoachMs Vanessa Loach

Imperial College Imperial College LondonLondon

Professor Frances Professor Frances GotchGotch

Professor Jonathan Professor Jonathan WeberWeber

Dr Nicola MackieDr Nicola MackieDr Peter HayesDr Peter Hayes

Dr Tristan BarberDr Tristan BarberMr Carl DeSouzaMr Carl DeSouza

Mr Ken LeggMr Ken LeggMs Miranda CowenMs Miranda Cowen

MRC Clinical Trials MRC Clinical Trials Unit LondonUnit LondonDr Sheena McCormackDr Sheena McCormackMs Elizabeth BrodnickiMs Elizabeth Brodnicki

Mr Patrick KelleherMr Patrick Kelleher

EMMESEMMESDr Leonid GibianskiDr Leonid Gibianski

Ms Carol SmithMs Carol SmithMs Kelley Loughran Ms Kelley Loughran

Mr Len DallyMr Len DallyMs Sophia PallasMs Sophia Pallas

OriginOriginMs Stephanie MiallMs Stephanie Miall

IAVIIAVIDr Claudia Schmidt Dr Claudia Schmidt

Dr Jill GilmourDr Jill Gilmour Dr Patricia FastDr Patricia FastMr Carl VerlindeMr Carl VerlindeMr Dani VoojisMr Dani Voojis

ALL VOLUNTEE

RS

Safe administration of DNA (pTHr.HIVA) and

MVA.HIVA to 169 HIV-1 uninfected

volunteers enrolled in Phase I/II trials in the UK.Additional SlidesAdditional Slides

XVI International AIDS ConferenceToronto, Canada.

Number of Volunteers who Number of Volunteers who received each vaccination as received each vaccination as

scheduled – Trial 006scheduled – Trial 006119 Enrolled Volunteers

40/40 2 mg DNA

EARLY

boost

LATE

boost

DNA

PRIMING39/39 0.5 mg DNA 40/40 Placebo

17/17 MVA 1/2 Placebo

OR

16/19 MVA 2/2 Placebo

17/18 MVA 2/2 Placebo

16/19 MVA 2/2 Placebo

17/17 MVA 1/1 Placebo

18/18 MVA 2/2 Placebo

16MRC Human Immunology Unit - WIMM - Oxford University

DemographicsAge #001 #003 #005 #006 #016

Median 34 34 43 36 43

Range 20-57 22-54 21-58 18-59 20-56

Gender

1

66

1812

28

53

666

0

20

40

60

80

#001 #003 #005 #006 #016

Female Male

Ethnicity155

65 30

30

60

90

120

150

Asian Black Caucasian Other

Asian Black Caucasian Other

17MRC Human Immunology Unit - WIMM - Oxford University

Distribution of Responses according to Age Trial #006

Distribution of MVA Responders according to Age

All MVA responders MVA-only Age n Mean Median n Mean Median all 19 36 35 5 32 29

> 40 5 52 51 1 52 52 40 14 27 27 4 28 27

All Responders (DNA-MVA= 9; MVA-only = 5) MVA-only excludes Responders in the DNA-placebo group

18MRC Human Immunology Unit - WIMM - Oxford University

Local & Systemic Reactions- Trial 006

(per volunteer during the 28 days post vaccination)Local Reactions Systemic Reactions

DNA MVA DNA MVA

Reactions post MVA were not altered by DNA priming

0

20

40

60

80

100

Placebo(N=40)

Vaccine(N=79)

Placebo(N=10)

Vaccine(N=101)

None Mild Moderate Severe

0

20

40

60

80

100

Placebo(N=40)

Vaccine(N=79)

Placebo(N=10)

Vaccine(N=101)

None Mild Moderate Severe

% %

19MRC Human Immunology Unit - WIMM - Oxford University

0

20

40

60

80

100

Placebo(N=63)

Vaccine(N=190)

Placebo(N=42)

Vaccine(N=377)

None Unlikely Possible Probable

0

20

40

60

80

100

Placebo(N=63)

Vaccine(N=190)

Placebo(N=42)

Vaccine(N=377)

Mild Moderate Severe

Non-Serious Adverse Events – Trial 006

(Excludes Local and Systemic Reactogenicity)

Severity Relationship

DNA DNAMVA MVA

%%

20MRC Human Immunology Unit - WIMM - Oxford University

HIV.A Peptides

HIV-1 gag CTL epitopes

p24 p17

Pool 1 Pool 2 Pool 3 Pool 4

Pool 90 (1+2+3+4)

Pool 9

Responder = volunteer with a positive response in at least one pool on at least one occasion.