Download andreas-suhrbier-qimr-berghofer-medical-research

Chikungunya virus vaccines?
Prof Andreas Suhrbier,
QIMR Berghofer
Medical Research Institute,
Brisbane, Queensland, Australia
Group Leader; Inflammation Biology QIMR B
-PRF NHMRC Australia
-Prof Griffith University & James Cook University
-Adj Ass Prof University of Queensland.
-Member; Australian Infectious Disease Res. Centre.
TEL +61 7 3362 0415
email [email protected]
www.qimrberghofer.edu.au/page/Lab/Inflammation_Biology/
QIMR-B, >600 scientists students & staff
“chikungunya” derived from the Makonde language
(Tanzania) means "that which bends up" referring to the
severe joint-pain-induced posture of afflicted individuals
- CHIKV is a biosafety level 3 pathogen (PC2 in Singapore).
- Category C Priority Pathogen by the National Institute of
Allergy and Infectious Disease (USA).
- US Army, CHIKV recognized as a potential bioweapon.
- Information export controlled under Defense Trade
Controls Act, 2012 (Australia)
Vadilal Sarabhai Hospital, Ahmadabad, India
ALPHAVIRUSES THAT CAUSE ARTHRITIC
DISEASE IN HUMANS
- Transmitted by mosquito (arboviruses)
- Single stranded positive sense RNA virus,
≈12 kb genomes.
- Symptomatic infections nearly always associated
with weeks to months polyarthritis/polyarthralgia.
Suhrbier, A. et al. 2012. Nature Rev. Rheumatol. 8, 420–429.
Virus
Chikungunya virus
Ross River virus
Barmah Forest virus
Sindbis virus family
Karelian fever
Ockelbo virus
Pogosta virus
O'nyong-nyong
Igbo Ora
Mayaro
Occurrence
Large sporadic epidemics every 2-50 years
Mean of ≈4,000 cases per annum in Australia.
Also an epidemic (1979/80) >60,000 cases
Mean of ≈ 1000 cases per annum in Australia
Rare (Karelia, West Russia)
Mean ≈30 cases per annum (Sweden)
Mean ≈140 (range 1-1282) cases p.a. (Finland)
Rare epidemics, >2 million cases in 1959-61
Small outbreaks (30-100 cases)
Largest ever CHIKV epidemic started 2004;
Estimated 1.4-6.5 million cases.
(Imported cases in ≈ 40 countries – not shown)
First ever
CHIKV
transmission
in Europe
Reached
PNG
2013
CHIKV has
been in
the USA,
1827/8
Approximate geographical locations of diseases associated with arthritogenic alphaviruses.
For CHIKV
disease, locations of documented large outbreaks are shown; epidemics prior to 1952 are shown in dashed lines and were initially
classified as outbreaks of dengue, but were likely to have been due to CHIKV. *Geographical locations of RRV and BFV diseases
overlap, with BFV restricted to the Australian mainland. ‡Main location of diseases caused by the Sindbis virus family. § O’nyongnyong virus disease outbreaks in 1959–1961 (East Africa), 1996–1997 (Uganda), 2003 (West Africa). Abbreviations: BFV, Barmah Forest
virus; CHIKV, chikungunya virus; RRV, Ross River virus
Suhrbier, A. et al. 2012. Nature Rev. Rheumatol.
>1 million
CHIKV cases in
the Americas
2014
First chikungunya case locally
acquired in the United States
reported in Florida July 2014
Reunion Island, French holiday
destination
CHIKV EPIDEMIC 2005/6,
REUNION ISLAND (FRANCE)
>250 deaths – often elderly with comorbidities
and very young
High attack rate
-266,000 cases of CHIKV disease were reported
(38% of the population).
(50% Grande Comore Island in 2005)
Rapid rise in case numbers
-increased to 130,000/month in 4 months
-45,000 cases during the week of 29 Jan, 2006.
Eurosurveillance,
Volume 11, Issue 34,
24 August 2006
Reunion cases
Imported cases
100’s of imported case in France
East-,Central-, and South-African (ECSA) phylogroup
The Indian Ocean/
Reunion Island
epidemic was
associated with a new
clade of CHIK viruses
Mutation allowed
efficient CHIK virus
transmission by
A. albopictus
Normal vector
A. aegypti
Schuffenecker et al Plos Med July 2006 3 ( 7 ) e263
© Landcare Research
Introduced
Native
Intercepted
Aedes albopictus;
a global invader
Tiger mosquito
An aggressive biter – the BBQ stopper
CHRONIC DISEASE
ELISA
based
serology
Virus
%
asymptomatic
Fever
Rash
Myalgia
Arthralgia
arthritis
CHIKV
5-18%
90%
40-50%
90%
>95%
RRV
55-75%
20-60%
BFV
?
50%
Very
15-40%
90%
common
80-100% 70-90%
SINV
ONNV
MAYV
8%
100%
80-100%
3-6 months
50-100% 50-80%
70-95%
40-60%
30-50%
40-80%
50%
95%
70%
60-100%
75%
50-90%
Disease characterised
by acute and chronic
symmetrical
polyarthritispolyarthralgia.
A Suhrbier, MC Jaffar-Bandjee, P Gasque.
2012. Arthritogenic alphaviruses - an
overview. Nat Rev Rheumatol. 8(7):420-9.
Prior to the recent epidemic,
chikungunya was not generally
considered to be a fatal illness
Reunion Island 260,000 CHIK cases
260 deaths (0.1%)
Mostly >75 years old
Ahmedabad (2006) 60,777 suspected
CHIK cases
2944 more deaths than in
same period of previous
year (4.8%)
Mavalankar et al 2008
For 610 atypical cases of CHIK:
222 were severe,
65 died.
Economopoulou et al 2009
Developed country mortality
0.01% – 0.1%
similar to influenza
CHIKV SEVERE MANIFESTATIONS (RARE)
Severe disease manifestation more
prominent in the elderly, the very
young & in patients with comorbidities e.g. hypertension, lupus
or cardiac disorders.
Haemorrhage & shock (occasional)
Rudd et al J Virol. 2012. 86(18):9888-98.
Mother to child transmission.
Reunion experience: 19 children born to
viraemic mothers in 7,504 pregnancies
• About half the children borne to viraemic
mothers became infected.
• About half the infected neonates
developed serious disease; haemorrhage,
DIC and/or cardiac and neurological
manifestations (often leading to permanent
disabilities)
Chronic
disease
The main burden of CHIKV disease is
chronic polyarthritis/polyarthralgia
Often several months occasionally > 1 year
(Hoarau et al. 2010. J Immunol 184:5914-27)
Viral arthritis is likely due to presence or persistence in joints of virus
or viral products that stimulate innate & cognate immune responses
Virus, viral antigens and/or viral RNA/DNA
found in arthritides caused by
Chikungunya virus (Hoarau et al. 2010)
Ross River virus
Rubella
Echovirus
Parvovirus B19
Caprine arthritis encephalitis virus
Avian reovirus
Adenovirus
CMV
EBV
Varicella
Virus infections can cause
auto-immune responses but
no good evidence these are
responsible for arthritic
disease or result in
autoimmune disease
Molecular mimicry – an
attractive hypothesis, but
little/no evidence.
A. Suhrbier, S. Mahalingham. The
immunobiology of viral arthritides. 2009.
Pharmacol Ther. 124(3):301-8.
Treatments
Rash – anti-histamine, calamine lotions (pruritis)
Indian J Dermatol. 2010 55(1): 64–67
Rheumatic symptoms/fever
NSAIDS/paracetomol - can provide relief, but often inadequate.
Rectal NSIADs (suppository)
Injectable NSAIDs in emergency settings?
Drug Saf. 1993 Nov;9(5):380-93.
NSAIDS plus steroids - some benefit - J. Rheumatol. 4, 94–101. (2009).
but must consider side effects of steriods
J Clin Rheumatol.10:326-30 (2004)
Chloroquine - ineffective - J. Med. Virol. 83, 1058–1059 (2011).
- Arthritis Rheumatol. 2014 Feb;66(2):319-26.
Methotrexate - benefit in some chronic patients (RA-like disease)
Expert Rev. Anti Infect. Ther. 8(9), 987–996 (2010). J Assoc Physicians India. 59:83-6 (2011)
No benefit in acute RRV mouse model
PLoS One. 2013. 8(8):e71146
VACCINES
Entity
Candidate CHIKV vaccines in development
Vaccine type
Pre-
Phase
Phase
Phase
Clinical
I
II
III
(1998)
 (2000)
USAMRIID
Live, attenuated (TSI-GSD-218, MRC-5 attunated strain of 15561)

Valnova/Karolinska Inst.
Live, attenuated (CHIKV-Δ5nsP3)

Takeda/UTMB
Live, attenuated (CHIKV/IRES)

Arbovax/NC State Uni
Live, attenuated (transmembrane deletion)

UTMB
Live, attenuated chimeric (various alphavirus backbones)

Themis Bioscience/Inst Pasteur
Live, vectored (measles virus)

 (2015)
Profectus/Yale/UTMB
Live, vectored (VSVΔG-CHIKV)

2016?
Karolinska Inst/CSIC Madrid
Live, vectored (MVA-CHIKV E1E226KE3)

Uni Wisconsin/Takeda
Live, vectored (MVA-CHIKV E2E3)

NIAID/Leidos Biomed
Virus-like particles (HEK293 production process)

TI Pharma/Wageningen Uni
Virus-like particles (Baculovirus/insect cell production process)

Merck
Virus-like particles (Baculovirus/insect cell production process)

Bharat Biotech
Inactivated (various strains, various methods)

Indian Immunological
Inactivated (formalin-treated 181/25 from US Army)

DRDE India
Inactivated (formalin-treated India 2006 isolate)

Nanotherapeutic Inc (from Baxter)
Inactivated (proprietrary adjuvant formulation)

Medigen
DNA (plasmid-launched 181/25 live attenuated)

DRDE India
Recombinant subunit (E coli expressed E1/E2)

National Inst. Virology, India
Recombinant subunit (E coli expressed E2)

Sementis (Australia)
Recombinant SCV (live recombinant attenuated vaccina)

 (2014)
2016?
Early CHIKV vaccines
• Formalin-inactivate vaccine; human
study (Harrison et al 1971 J Immunol
107; 643-647).
• Live-attenuated vaccine. Phase II
human trial (Edelman et al., 2000).
Side effects included arthralgia
Lancet 2014; 384: 2046–52
• 25 participants were enrolled
10 μg (n=5), 20 μg (n=10), and 40 μg (n=10).
• Neutralising antibodies were detected in all dose groups after
the second vaccination
• All injections were well tolerated, with no serious adverse
events reported.
Manufacturing costs? Memory? Adjuvant not used but could provide benefits.
Lancet Infect Dis 2015; 15: 519–27
• 42 participants to receive the low dose (n=12), the medium
dose (n=12), or the high dose (n=12)
• Second vaccination resulted in a 100% seroconversion for all
participants
• No vaccination-related serious adverse events were recorded.
Overcoming measles immunity especially in recently vaccinated children requires
dose increases increasing risk of adverse events
CHIKV vaccines – some considerations
Sporadic epidemics (2-50 years), usually self limiting disease, mortality low and
usually restricted to elderly and patients with comorbidities.
Protective correlate
• Antibodies are protective (T cells have a minor role)
• T cells also associated with arthritis (induction of CD4 T cells in the
absence of good antibodies in mouse models promotes arthritis
upon challenge – Poo et al Plos NTD 2014)
Market for CHIKV vaccines?
Tourists?
Army?
General population?
Desirable for an epidemic setting
• A single shot, rapid onset of protective immunity.
• Capacity for rapid manufacture and deployment
• Low cost, highly stable (afflicted countries often resource poor)
QIMR Berghofer
7 suite
Biosafety level 3
floor with
equipment
animal houses
& insectaries.
Large PC2 mouse breeding
facility 14,000 cage capacity
>140 genetically
modified mouse strains
Research Innovation 2012;
Battling viral rheumatism
Viraemia
6
5
4
3
2
1
0
0
1
2
3
4
5
6
Days post CHIKV inoculation
Control
Reunion
10
Foot width x breadth,
mm2 ± SE
Viraemia, log10CCID50/ml ± SE
Adult wild-type mouse model of chikungunya virus
infection (viraemia) and disease (foot swelling/arthritis)
Arthritis
Control
9
8
7
0
2
4
6
8
10
12
Days post CHIKV inoculation
Reunion Island
isolate
Female C57BL/6
mice > 6 weeks old
14
Dominance of monocytes/macrophages and NK cells in the
swollen feet of CHIKV infected mice
FACS
Cells
FACS markers
% of cells
7 days post infection
± SE
Monocytes
CD11b+, F4/80lo/-
45 ± 9.9
Macrophages
NK cells
CD4 T cells
CD8 T cells
B cells
F4/80+, CD11b+
NK1.1+
CD3+, CD4+
CD3+, CD8+
CD19+
CD11c+, F480-,
NK1.1B220-, PDCA1CD11c+, B220+,
PDCA1+
21 ± 4
16.3 ± 6
5.2 ± 1.3
1.2 ± 0.2
4.6 ± 2
Conventional
DC
Plasmacytoid
DC
6.4 ± 1.1
3.8 ± 2.1
Virus strain LR2006-OPY1
injected s.c. into the foot
Air hole
J. Virol. Meth. 1995. 52:51-54
Many virus
isolates
contaminated
with
mycoplasmas
MycoAlert™
Mycoplasma
Detection Kit
MYCOPLASMA
NO MYCOPLASMA
Hela and Hoechst staining
Results in < 20 min
Grow virus prep on LCLs (non permissive for alphavirus infection for a week) prior to testing
ENDOTOXIN CONTAMINATION
(often in FCS, glassware, trypsin, sucrose etc)
“High sensitivity” endotoxin assay detection limit
Clear inhibition of alphavirus infection
in macrophage cell lines
Johnson.et al
2005. J Biol
Chem.
280(6):4037-47
65C 30 mins to inactivate virus
RAW264-HIV-LTR-LUC cell line
Detection sensitivity 5-10 pg/ml E. coli LPS
Examples of the evaluation of vaccines in
the adult wild-type CHIKV mouse model
1. Simple inactivated whole virus vaccine
Gardner et al. J Virol. 2010 84(16):8021-32
2. rBaculovirus VLP vaccine Metz et al PLoS NTD 2013;7:e2124
A collaboration with Wageningen University, The Netherlands.
3. rAdenovirus vaccine
Wang et al 2011 Vaccine 29;2803–2809
A commercial collaboration with GenPhar Inc., USA.
4. rSementis Copenhagen Vector (SCV)
A commercial collaboration with Sementis, Australia
4. Foroderm transcutaneous immunisation
A collaboration with University Queensland, Australia
Model adopted by EU Integrated CHIKV Research Program, e.g.
• rMVA vaccine (García-Arriaza et al. J Virol. 2014. 88(6):3527-47)
• Attenuated vaccine (Hallengärd et al. J Virol. 2014 88(5):2858-66)
Metz et al, 2013
CHIKV VLP production in Sf21 insect cells
VIRAEMIA
Gorben Pijlman, Wageningen University
ARTHRITIS
Single shot:
Complete
protection
against
viraemia
Complete
protection
against
arthritic
disease
Rapid high volume production
9
8
SCV -CHIK
Control vector
PBS
7
6
5
4
3
2
1
0
0
1
2
3
4
Day post challenge
5
% increase in foot swelling (arthritis) + SE
Log10 CCID50/ml ± SE
The Sementis Copenhagen Vector (SCV) is a genetically
attenuated, live virus vaccine vector, based on the
Copenhagen strain of the Vaccinia Virus
80
SCV -CHIK
Control vector
PBS
70
60
50
40
30
20
10
0
0
2
4
6
8
10
12
14
16
Day post challenge
Complete protection from viraemia and arthritis
by single vaccination with rSCV-CHIKV vaccine
SCV can be produced from a
biotechnology “friendly” custom cell
line under serum and protein free
conditions in bioreactors
COI: A Suhrbier is on the SAB of Sementis
ForodermTM : Novel Drug Delivery Technology
Dr Tarl Prow et al University Queensland, Australia
Potential for self administration
Foroderm + inactivated CHIKV
120
NEGATIVE CONTROLS
Topical inactivated CHIKV
Topical inactivated CHIKV+QuilA
Foroderm plus QuilA
POSITIVE CONTROL
Sub. cut. inactivated CHIKV
110
100
90
80
70
60
50
40
Arthritis
30
20
10
0
0
Viraemia, log10 CCID50 ml + SE
% increase in foot swelling (arthritis) + SE
Single application of
inactivated CHIKV vaccine
with Foroderm provides
complete protect against
Arthritis and Viraemia
5
10
15
20
30
25
35
Days post challenge
6
5
Viraemia
4
3
2
1
0
0
1
2
3
4
Days post challenge
5
6
Phase II/III testing?
Sporadic epidemics (2-50 years)
Epidemics
Year
No. of cases
Tanzania
1952
?
Central East Africa
1959-62
≈2 m (O'nyong'nyong)
Thailand
1962
≈40,000
India
1963/4
>100,000
Outbreak size
Vietnam Myanmar
1975
?
and location
Indonesia
1982
?
unpredictable
Reunion Island
2005-2006
>250,000
India/Asia/Caribbean/S America
2006-2015
1.4-6.5 m
By the time ethics approval given, trial infrastructure set up and
volunteers vaccinated. Epidemic has passed or does not arrive.
USA FDA unlikely to give approval on animal data alone
CAN EMERGING ECONOMIES DO THEIR OWN APPROVAL ?
E.g. Brazil ? India ?
Mosquito control
• Use real insect repellent (eg DEET)
• Wear long sleeved clothes
• Uses mats, coils, screens indoors
• Use impregnated bed nets
• Spraying
•Remove peri-domestic breeding sites
•Add fish to ponds and water tubs
Make mosquitos less able to
transmit CHIKV with Wolbachia
Wolbachia is a
genus of bacteria
which infects
insects
Release of
Wolbachia
infected
mosquitos into
the wild
population results
in spread of the
Wolbachia
infection into the
population
Wolbachia
infected
A . aegypti
mosquitos
replicate
dengue virus,
chikungunya
virus & yellow
fever virus less
efficiently
RT PCR
+ Wol
- Wol
Hurk et al 2012 PLoS Negl Trop Dis 6(11): e1892.
Walker et al., 2011. Nature 476, 450–453.
Scot ONeil
Hoffmann et al 2011. Nature. 2011;476:454-7.
RA
and UP regulated in RA patients
282 genes UP regulated in CHIKV arthritis
Healthy
-2
0
SD from mean
Improving treatment options
Surprising overlap in
inflammatory signature
between CHIKV and
rheumatoid arthritis.
Drugs being
used/developed for RA
may find application for
treatment of CHIKV.
(Nakaya et al, 2012. Arth Rheum
64 (11):3553-63)
Heat map showing the relative expression levels of the 282
2
4.5 5 8 7 7.5
RA inflammation score
up-regulated genes in CHIKV infected mouse feet et that were
enriched in RA patients. Columns represent the 5 control, and
5 RA samples in the study and their respective inflammation
scores.
QIMR-B
Joy Gardner
Penny Rudd
Itaru Anraku
Thuy T Le
Lee Major
Wayne A. Schroder
Ecole Nationale Vétérinaire, Nantes, France
Thibaut Larcher et al
Commissariat à l'Énergie Atomique (CEA), Paris
Karine Labadie, Pierre Roques et al
University of Texas Medical Branch
Stephen Higgs et al
University of Queensland
Tarl Prow
University of Wageningen
Roy Hall
Gorben Piljman et al
Alex Khromykh
Griffith University
Suresh Mahalingham
Nestor Rulli
Funding
National Health & Medical Research Council, Australia
Australian Infectious Disease Research Center
Queensland Tropical Health Alliance
Université de la Réunion
Hôpital Félix Guyon
La Réunion, France
Marie-Christine Jaffar-Bandjee
Philippe Gasque
Emory Vaccine Center
Helder Nakaya, Bali Pulendran
Thanks to
• Luis Mateo and Rebecca Pawliw
(Alere, Brisbane, Australia) for supply
of purified inactivated CHIKV.
• Clay Winterford (QIMR) for
histology/immunohistochemistry
Information resources
French guidelines for the management of chikungunya (acute and persistent
presentations). November 2014.
Simon et al. Med Mal Infect. 2015 Jul;45(7):243-63.
Queensland chikungunya management plan 2014–2019.
https://www.health.qld.gov.au/cdcg/documents/chikungunyamanagement-plan.pdf (accessed 14/7/15).
Chikungunya virus. Centers for Disease Control and Prevention.
http://www.cdc.gov/chikungunya/fact/index.html (accessed 17/6/15).