Adolfo García-SastreIcahn School of Medicine at Mount Sinai, New York

Influenza Universal Vaccines

P01AI097092

Disclosure: Some of these studies are funded by a GSK research agreement. AG-S is inventor in patents on universal flu vaccines.

INFLUENZA VIRUSES

PAx

‘40 ‘50 ‘60 ‘70 ‘80 ‘00‘901918

H1N1

H2N2

H1N1

H3N2

A

B

EPIDEMIOLOGY OF HUMAN INFLUENZA

VIRUSES

‘10

1957

1968

1977

pH1N1

2009

INFECTIONS IN HUMANS WITH AVIAN AND

SWINE INFLUENZA A VIRUSES

H5N1 H9N2 H5N1 H5N1H7N7

‘40 ‘50 ‘60 ‘70 ‘80 ‘00‘901918

H1N1

H2N2

H1N1

H3N2

A

B

‘10

1957

1968

1977

pH1N1

2009

H7N9H3N2v

Evolution and spread of flu viruses

Spread of the pandemic H2N2 virus, 1957

Source: CDC ILI and Vaccine Distribution Data

Pandemic H1N1 cases and vaccinations in US Sept 2009 – May 2010

0

20 000 000

40 000 000

60 000 000

80 000 000

100 000 000

120 000 000

140 000 000

0

1

2

3

4

5

6

7

8

9

03/09/2009 03/10/2009 03/11/2009 03/12/2009 03/01/2010 03/02/2010 03/03/2010 03/04/2010 03/05/2010

Nu

mb

er

of

H1N

1 V

accin

e S

hip

ped

% o

f V

isit

s f

or

ILI ILI Shipped

Vaccine

Universal flu

vaccines?

Neutralization of influenza viruses

Nat Struct Mol Biol. 2009 Mar;16(3):233-4.

HA1

HA2

Neutralizing antibodies

Hemagglutinin subtypes

HA-STEM BASED UNIVERSAL FLU VACCINES?

1. Use of headless constructs

Strategies to overcome HA-head immunodominance

Sagawa et al. (1996). J Gen Virol 77 ( Pt 7), 1483-1487.

Steel et al (2010). MBio 1.

Bommakanti et al (2012). J Virol 86, 13434-13444.

Mallajosyula et al. (2014). Proc Natl Acad Sci USA 111, E2514-2523.

Lu et al. (2014). Proc Natl Acad Sci U S A 111, 125-130.

Wohlbold et al. (2015). Vaccine 33, 3314-3321.

Impagliazzo et al. (2015). Science

Yassine et al. (2015). Nat Med 21, 1065-1070

UNIVERSAL FLU VACCINES?

2. Repeated vaccination with influenza

virus chimeric HA vaccines induce

protective antibodies against multiple

subtypes of influenza virus.Irina Margine Randy Albrecht

Florian Krammer

Rong Hai Patrick Wilson

Gene Tan S.A. Andrews

Peter Palese Jon Runstadler

cH4/3 DNA cH5/3 protein

boost

H3 protein

boost

Shanghai

(H7N9)

challengeControl groups:

cH4/3 DNA + BSA + BSA

naïve (neg. contr.)

matched vaccine (pos. contr.)

4 weeks3 weeks3 weeks

Induction of protective levels of stalk-reactive

antibodies using chimeric HA constructs in mice

Proof of principle

cH4/3 DNA cH5/3 protein

boost

H3 protein

boost

Shanghai

(H7N9)

challengeControl groups:

cH4/3 DNA + BSA + BSA

naïve (neg. contr.)

matched vaccine (pos. contr.)

4 weeks3 weeks3 weeks

Induction of protective levels of stalk-reactive

antibodies using chimeric HA constructs in mice

Proof of principle

cH4/3 DNA cH5/3 protein

boost

H3 protein

boost

Shanghai

(H7N9)

challengeControl groups:

cH4/3 DNA + BSA + BSA

naïve (neg. contr.)

matched vaccine (pos. contr.)

4 weeks3 weeks3 weeks

Y

Induction of protective levels of stalk-reactive

antibodies using chimeric HA constructs in mice

Proof of principle

cH4/3 DNA cH5/3 protein

boost

H3 protein

boost

Shanghai

(H7N9)

challengeControl groups:

cH4/3 DNA + BSA + BSA

naïve (neg. contr.)

matched vaccine (pos. contr.)

4 weeks3 weeks3 weeks

Y

Induction of protective levels of stalk-reactive

antibodies using chimeric HA constructs in mice

Proof of principle

cHA vaccine protects against

challenge with novel H7N9 virus

cHA vaccine protects against

challenge with H10 and H3 viruses

cH4/3 DNA + cH5/3 protein + H3 protein cH4/3 DNA + cH5/3 protein + cH7/3 protein

Titers in mouse lungs, day 3 postinfection

Abs mediate protection

H3N2

H11

H13

H16

H1

H2

H5

H6

H17

H8

H12

H9

H7

H15

H10

H3

H4

H14

GROU

P 1

GROU

P 2

0.04

Targeting group 1 HA viruses

cH9/1 DNA cH6/1 protein cH5/1 protein

Control groups:

cH9/1 DNA + BSA + BSA

matched vaccine (pos. contr.)

YPR8 H1N1

FM1 H1N1

pH1N1

H5N1

H6N1

challenge

Induction of protective levels of stalk-reactive

antibodies using chimeric HA constructs in mice

Proof of principle

Vaccination with cHA constructs protects

from pH1N1 (A/Netherlands/602/09) challenge

positive control (matched inactivated)

cH9/1 DNA + cH6/1 protein + cH5/1 protein

cH9/1 DNA + BSA +BSA

Similar results for A/PR/8/34 H1N1 and A/FM/1/47 challenges

positive control (matched inactivated)

cH9/1 DNA + H1 protein/cH6/1 protein + cH5/1 protein/H1 protein

cH9/1 DNA + BSA +BSA

cHA constructs protect mice from

heterosubtypic challenge

H5N1 challenge H6N1 challenge

cH5/1 (H5 challenge) or cH6/1 (H6 challenge) protein was replaced by full length

H1 protein to exclude head-based protection

ELISA reactivity to Cal09

(pH1N1) protein

Protection is antibody mediated

cH9/1 + cH6/1 + cH5/1

cH9/1 + BSA +BSA

naïve serum

Naïve

Positive control

vector +BSA+BSA

cH9/1 + cH6/1 + cH5/1

Passive transfer of serum

protects from viral challenge

Days post challenge

Prime–Boost cHA vaccines

based in

LAIV and IIV platforms

Florian Krammer, Raffael Nachbagauer,

Adolfo García-Sastre, Peter Palese and Randy A. Albrecht

cH8/1 - LAIV cH5/1 - IIV

cH8/1 - IIV cH5/1 - IIV

Boost Boost

“cH8/1 LAIV- cH5/1 IIV”

TIV

Prime

B-cH9/1

B-cH9/1

B-cH9/1 Mock Mock

Mock

Ferret vaccination groups (n=4)LAIV is based on the Ann Arbor backbone

“cH5/1 IIV- cH5/1 IIV”

“Prime-only”

“TIV”

“Naive”

Prime–Boost cHA vaccines based in

LAIV and IIV platforms

po

st-

TIV

po

st-

pr i

me

po

st-

pr i

me

po

st-

cH

8/1

IIV

po

st-

cH

5/1

IIV

po

st-

pr i

me

po

st-

cH

8/1

LA

IV

po

st-

cH

5/1

IIV

1 0 0 0

1 0 0 0 0

1 0 0 0 0 0

S ta lk -tite r (c H 6 /1 H A )

en

dp

oin

t ti

ter

c H 8 /1 L A IV - c H 5 /1 I IV

T IV

P r im e o n ly

c H 8 /1 I IV - c H 5 /1 I IV

Induction of HA stalk-specific antibodies (ELISA)

*No detectable HI titers following vaccination

Induction of NA-specific antibodies (ELISA)

Na

ive

TIV

Pr i

me

-on

ly

cH

8/1

IIV

- c

H5

/1 IIV

cH

8/1

LA

IV -

cH

5/1

IIV

1 0 0

1 0 1

1 0 2

1 0 3

1 0 4

1 0 5

1 0 6

1 0 7

T r a c h e a

pla

qu

e f

orm

ing

un

its

/gra

m

Na

ive

TIV

Pr i

me

-on

ly

cH

8/1

IIV

- c

H5

/1 IIV

cH

8/1

LA

IV -

cH

5/1

IIV

1 0 0

1 0 1

1 0 2

1 0 3

1 0 4

1 0 5

1 0 6

1 0 7

N a s a l tu rb in a te s

pla

qu

e f

orm

ing

un

its

/gra

m

Na

ive

TIV

Pr i

me

-on

ly

cH

8/1

IIV

- c

H5

/1 I

IV

cH

8/1

LA

IV -

cH

5/1

IIV

1 0 0

1 0 1

1 0 2

1 0 3

1 0 4

1 0 5

1 0 6

1 0 7

B r o n c h u s

pla

qu

e f

orm

ing

un

its

/gra

m

Viral titers in tissues following H1N1 challenge infection, day 4

CONCLUSIONS

LIVE ATTENUATED FOLLOWED BY INACTIVATED

CHIMERIC HA VACCINES INDUCE HA STEM AND NA

ANTIBODIES,

AND HIGH LEVELS OF PROTECTION AGAINST

HETEROSUBTYPIC CHALLENGE IN FERRETS

Acknowledgements

Randy AlbrechtMichael Schotsaert

Angela ChoiJuan Ayllon

Raffael Nachbagauer

Florian KrammerPeter PaleseRafi Ahmed

Patrick Willson