Translating research into practice Immunisation for genital viral infections Ian Frazer, Research Director, The Translational Research Institute Brisbane, Australia
Translating research into practiceImmunisation for genital viral infections
Ian Frazer, Research Director, The Translational Research Institute Brisbane, Australia
Infection causes >20 % of cancerAnd we’re still finding new ones.% Infectious agent Associated Cancer
Viruses
5.2 Papillomavirus Anogenital, oropharyngeal
3.9 Hepatitis B virus Hepatocellular
1.0 Epstein Barr Virus Nasopharyngeal Ca, Lymphoma
2.9 Hepatitis C virus Hepatocellular Ca
0.9 HIV Potentiates viral cancers
<0.2
HTLV -1 T cell leukemia
<0.2
Merkel polyomavirus
Merkel’s disease
<0.2
HHV -8 Kaposis sarcoma
Other
5.5 H. Pylorii Gastric
<0.2
Parasites Bladder, ?HCC
Parkin DM et al Int J Cancer 118, 3030 (2006)
NB Infections common, cancer rare consequence
HPV vaccine development:A 15 year process
Cancer associated Human
Papillomaviruses(~1980)
100% of cervical cancer and 30% of
head and neck cancer caused by “high risk” HPVs
(16,18)
Virus Like particles1
(~1990)
Major L1 capsid protein
expressed in vitro self
assembles to VLPs
HPV vaccines(~2005)
- VLPs+ adjuvant - Neutralising Ab- Protection >8yrs-Cervarix - HPV 16, 18-Gardasil – HPV 6,11,16,18
1:J. Zhou, X. Y. Sun, D. J. Stenzel, and I. H. Frazer. Virology 185 (1):251-257, 1991.
Jian Zhou
Zur Hausen
Nine steps to translate HPV research into practical outcomes
• Define the problem – HPV and cancer Zur Hausen
• Develop the science –recombinant VLPS Jian Zhou• Find commercial partners
Merck/GSK• Scale up the manufacture
Merck/GSK• Complete safety and efficacy trials Many
groups• Educate the profession, govt, the public Ongoing• Confirm field efficacy • Improve the product
– 9 valent vaccines, 2 dose regimens• Encourage deployment globally• Develop effective immunotherapy
Disclosure of conflict of interest
Dr Ian Frazer and the University of Queensland benefit financially from commercial sale of the prophylactic HPV vaccines discussed in this talk
Genital warts in young vaccinated women in Australia
Ali, H. Et al BMJ 2013;346:f2032 doi: 10.1136/bmj.f2032
Geniital Warts have gone away in their unvaccinated male partners
Ali, H. Et al BMJ 2013;346:f2032 doi: 10.1136/bmj.f2032
Differences in HPV genoprevalence between prevaccine and postvaccine
populations.
Tabrizi S N et al. J Infect Dis. 2012;infdis.jis590
Latest Australian data for PAP samples 90% reduction in HPV16, 18 in < 30yrs
There is disparity between HPV vaccine programs and cervical cancer prevalence
http://www.indexmundi.com/blog/wp-content/uploads/2013/05/HPV-vaccine-infographic.jpg
Vaccine delivery logistics in Vanuatu:
staffing, and a cold chain Vanuatu->50 islands~250,000 people~25 doctors-91 parliamentarians!-1 vaccine fridge-0 reliable electricity
500 well women over 30
Bx result Number
Cancer 5
CIN 3 17
CIN 1 23
Education in Vanuatu: parents, children, staff, and
government
Parents and Kids under the Banyan Tree- north Efate
15
When vaccine is available, vaccination programs in
subsistence economies can be as successful as
Australia
State Age Dose 1 Coverage (%)
Dose 1 immunised receiving
Dose 2 (%) Dose 3 (%)
Australia 12-18 (Opt out)
~ 90% 96 86
Australia 18-25 (Opt in) ~80% 82 50
Vanuatu 10-12 (Opt out)
~80% 98 93~80% coverage for bivalent HPV vaccine for 10-12 year olds in 2008, and 2013Vaccine through Australian Cervical Cancer Foundation: delivery by government
• E6/E7 – recombinant bacterial
fusion of HPV16 E6 and E7 proteins in 8M Urea
• ISCOMATRIX® adjuvant 120µg/dose– Quillaia saponin based
adjuvant– typically 40nm cage
like structures– promotes both
humoral and cell mediated immunity
ISCOMATRIX® adjuvant
HPV 16 vaccine
100nm
Immunotherapy for HPV associated cancer
Immunotherapy: can we get rid of existing HPV infection
Colposcopy- no change
Pre- and post- vaccination
1 quadrant 9
2 quadrants 11
3+ quadrants 10
Frazer, I. H., M. Quinn, et al. (2004). Vaccine 23(2): 172-181.
3317CIN 1
12223CIN 2,3
NilCIN 1
CIN 2,3
Post vaccination histology
NPre vaccination
histology
Histology- no change
Immunology – a nice response 20mg (3x)
Placebo (1x or 3x)
60mg (3x)
200mg (1x)
DTH
(m
m)
Pre Post Pre Post
0
10
20
30
40
50
100>200
0
10
20
30
40
50
100>200
0
10
20
30
40
50
100>200
0
10
20
30
40
50
100>200
• Antibody +++ • DTH +++• Cytotoxic +++ T cells
Modelling epithelial cancer immunotherapy
Keratinocyte-immune cell interactions
• Antigen expressed only in basal keratinocytes• Stable expression of antigen over time without tumor
related phenomena• Grafts expressing some antigens (eg OVA) reject
spontaneously, • Grafts expressing others (eg HPV E6, E7) don’t
K14 transgenic
Nontransgenic control
Allo matched nontransgenic
RECIPIENT
DONORS
How does inflammation work?A skin resident immunocyte locally inhibits
E7 specific effector T cell function
Not reproduced by:Depletion of CD25+ regulatory cells from graft (Mab), Elimination of FoxP3 positive cells from graft ( DEREG
mouse)Therefore – another regulatory T cell population exists in skin
No immune cells (rag-/-) in E7 graft
immune cells in E7 graft
Mattarollo, S. R., A. Rahimpour, et al. (2010). The Journal of Immunology 184(3): 1242-1250.
NKT CELLS AND IFN-γ ARE KEY REGULATORS OF LOCAL T CELL FUNCTION
Steve Mattarollo, Rachel DeKluyver, Christina Gosmann
The inhibitory cell is an NKT cell.
Mattarollo, S. R., A. Rahimpour, et al. (2010). "Invariant NKT cells in hyperplastic skin induce a local immune suppressive environment by IFN-gamma production." The Journal of Immunology 184(3): 1242-1250.
Conclusion: NKT cells locally inhibit effector function of cytotoxic T cells
NKT cells
No NKT cellsNote – inhibition of rejection local to NKT replete graft
Footnote:Of mice and (wo)men!
Parameter
In mice In humans
IDO In E7tg + Inhibits rejection In CIN a
IL-17 In E7tg + Inhibits rejection In CIN a
IFN-g In E7tg + Inhibits rejection In CIN a
Mast cells In E7tg + Inhibits rejection In CIN a
IL1Ra In E7tg + Inhibits rejection In CIN b
NKT cells Inhibit rejection
a: Frazer Lab Datab: Fujiwaki R Gynecol Oncol. 2003 Apr;89(1):77-83
Using polynucleotides as vaccinesMaking the body’s cells do the work
Codon modification = better responses
0.0
0.5
1.0
1.5
1:50 1:200 1:800 1:3200Serum Dilution
CODON MODIFIED (1 shot)
CODON MODIFIED (2 shots)
NO MODIFICATION
IMM
UN
E R
ES
PO
NS
ETO
VA
CC
INE
Another improvement Ubiqitination = better cellular immunity
L 1 mod E7Ubi
L 1 mod E7H6L1 E7
UH6L1E7
Mixed polynucleotide vaccines have the immunogenicity of both
components
0
25
50
75
100
MIX (LN)MIX (S)Control (LN)Control (S)
A
CT
LIF
N s
pots
/ 10
6ce
lls
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1:50 1:200 1:800 1:3200
B
Co
nfo
rma
tio
na
la
nti
bo
die
s
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1:50 1:200 1:800 1:3200
C
Ab
to
de
na
ture
d L
1
HSV- 2 Vaccine Development
o Based on glycoprotein D
o Initial gD2 constructs:
• control (wt CU gD2)
• codon modified (O1, O2 and O3 gD2)
• codon de-optimised (W gD2)
o Initial studies identified O2 as best performer
o Made ubiquitinated version of O2
Challenge Survival
50 X LD50
0 1 2 3 4 5 6 7 8 9 1011121314151617181920210
10
20
30
40
50
60
70
80
90
100
days post-infection
surv
ival
(%
)
500 X LD50
0 1 2 3 4 5 6 7 8 9 1011121314151617181920210
10
20
30
40
50
60
70
80
90
100
O2-gD2
O2-UgD225-331
gD mix
W-gD2
empty vector
TK-
days post-infection
HSV-2 Vaccine Phase 1 Study
o Safety, Tolerability & Immunogenicity Endpoints
o Open Label, Multiple Dose Escalation
o Doses of 10µg, 30µg, 100µg, 300µg, and 1mg
o Intradermal injections (Days 0, 21 and 42)
o 22 healthy sero-negative 18-45 yr old subjects
enrolled from 59 volunteers
HSV-2 Vaccine Phase 1b Study
o 40 18-45 yr old subjects with recurrent culture/PCR
proven HSV-2
oRandomised, blinded, 1mg Dose or placebo
oIntradermal injections (Days 0, 21 and 42)
oSafety, Tolerability & Immunogenicity primary
endpoints
oViral shedding before (2 months) and after (2 months)
vaccine secondary endpoint