GRF 2nd One Health Summit 2013: Presentation by Veronika Von Messling, Paul-Ehrlich-Institute

1. Paul-Ehrlich-Institut
Bundesinstitut für Impfstoffe und biomedizinische Arzneimittel
A Morbillivirus Vaccine Vector
Expressing Influenza HA and NP
Proteins Induces Robust Humoral
and Cellular Immune Responses
Veronika von Messling, DVM, PhD
One Health Summit, Davos, November 20, 2013

2. Influenza Virus Epidemiology
 Influenza A viruses infect swine,
horses, seals, and a large variety of
birds as well as humans
=> Eradication is not feasible
 All subtypes of influenza A viruses
(1-17 HA and 1-10 NA) are found in
aquatic birds.
 A pandemic occurs when a “new”
subtype adapts to humans.
Challenge:
Lakadamyali et al., PNAS 2003
Currently available HA/NA based
vaccines only protect against
antigenetically matched viruses.

3. Ferret Model for Respiratory
Viruses
• Signs and severity of disease closely resemble
human disease
• Strong similarities to humans in lung
physiology, airway morphology and cell types
present in the respiratory tract
• Sialic acids are predominantly -2,6-linked
(influenza)
• Small animal model that can be
accommodated by most animal facilities
• Immunological reagents are being developed
• Ferret genome project is ongoing

4. Seasonal H1N1 Pre-Infection
Protects Ferrets at Least as
Well as an Inactivated H5N1
Vaccine
Groups:
1. control:
2. H1N1inf:
PBS - 2 x i.m. injection (n=3 per time point)
intranasal infection with 105 infectious doses of H1N1
USSR/90/77 (n=5-6 per time point)
3. H5N1vac: Formalin-inactivated H5N1 vaccine - 2 x i.m. injections
(4 µg H5 HA equiv.; Viet Nam 1203/04; n=5-6 per time
point)

5. l
1i
nf
ac
ro
1v
1N
5N
H
H
day 3
l
1i
nf
ac
ro
1v
1N
5N
H
H
nt
10
co
nf
nf
ac
1i
1v
l
nf
ro
1i
nt
1N
5N
H
H
co
1N
ac
l
nf
ro
1v
l
ac
ro
1i
nt
1N
5N
H
H
nt
1v
co
H
5N
*
nt
nf
1i
ac
l
nf
ro
1v
nt
1N
5N
H
H
co
1i
ac
H
co
1000
co
l
ac
1i
1v
1N
5N
nf
ro
1i
1N
1v
l
*
H
H
day 1
*
nt
1N
5N
H
H
ro
1000
co
H
ac
l
nt
10000
1v
ro
10000
co
1
100000
5N
nt
1
100000
H
co
log10 TCID50
100000
10000
*
6 months
100
10
100
*
1000
100
10
1
*
day 6
12 months
*
18 months

6. Can we design a vaccine
reproducing this cross-protection?
Expectations:
Milder and shorter disease.
Reduced viral load = less transmission.
Requirements:
Induction of humoral and cellular immune responses.
Immune responses against HA as well as conserved
viral proteins.

7. Live-Attenuated Morbillivirus Vaccines
Induce Long-Lasting Protection
Morbilliviruses as Vaccine Platforms
Introduction of immunogenic proteins from other
pathogens in the morbillivirus genome
Expression of the “foreign” protein in the
highly immunogenic morbillivirus context
Cellular and humoral immune response to the
morbillivirus + “foreign” antigen

8. Generation of a CDV Vaccine
Carrying Influenza Antigens
CDV Vaccine Strain Onderstepoort (OS)
OS expressing NP and HA from H1N1 USSR/90/77 (OS NPHA)
OS
OS NPHA

9. Experimental Design
Immunization
Challenge
Sacrifice
E
E
E
ELISpot
serum
0
7
10
14
21
28
31
days after vaccination
Immunization:
5x105 infectious doses of OS or OS NPHA, i.m. injection
(n=9/virus)
Challenge:
intranasal infection with 105 infectious doses of n=3 of each
group/virus):
a) H1N1 USSR/90/77 (homologous virus)
b) pH1N1 Mx/09 (heterologous – same subtype)
c) H3N2 Port Chalmers/1/71 (heterologous – different
subtype
Sacrifice:
3 days post-challenge to collect nasal turbinate and lung
tissues for titration

10. Both Viruses Induce a Robust
Immune Response Against CDV
SFC/106 MNCs
VN Titer
Day 0
Day 10
Day 28

11. Only OS NPHA Elicits Also a
Response Against Influenza
SFC/106 MNCs
HAI Titer
Day 0
Day 10
Day 28

12. Vaccination with OS NPHA Reduces
Viral Load after Homologous and
Heterologous Challenge
H1N1-matched
Nasal Turbinate Titer
Day 3
H1N1-mismatched
TCID50/mg tissue
Body temperature C
H3N2
Days post-infection
*
**
**

13. How Does This Relate to One
Health?
• Influenza is a prototypical zoonotic disease –
successful control will involve addressing
human and animal hosts
• Vaccination is the most cost-effective and
efficient public health measure against
infectious diseases.
• Reverse engineering of vaccines – i.e.
identifying correlates of immunity and designing
a vaccine that induces this response – allows
the development of tailor-made vaccines.

14. Acknowledgements
INRS-Institut ArmandFrappier, Laval
National Microbiology
Laboratory, Winnipeg
Ronan Rouxel
Isabelle Meunier
Xiao Xiang Wong
Darwyn Kobasa
Michael Gray
Jason Gren
Shane Jones
Betty Luy
Gary Kobinger
Funding
CIHR, CFI, Armand-Frappier
Foundation, Duke-NUS

15. Skin Rash

16. CDV Spreads Through the Host
von Messling et al., PNAS 2004

17. Viral Take-Over of Lymphatic Tissues
Precedes Epithelium Invasion
Thymus
Spleen
10x
Bladder
10x
Oesphagus
10x
10x
4 d.p.i.
8 d.p.i.
14 d.p.i.
lymphatic cells
epithelial cells
Rudd et al., JVI 2005