Measles virus vaccine induces oncolysis of
tumor cells and activates immune responses.
Marc Grégoire*, J.-F. Fonteneau, N. Boisgerault, J.-B. Guillerme and F. Tangy¤.
* INSERM, Nantes, F-44000, France.
¤ Pasteur Institute, Viral Genomics and Vaccination, Paris, France
Oncolytic
virus
Ideal replicating oncolytic virus:
Russell SJ, Nature biotechnol, 2012
Vacchelli E et al, Oncoimmunology, 2013
- Kills efficiently tumor cells (apoptosis)
Infection
Healthy cells
- No toxicity (genetically stable, minor side effects)
- High viral concentration production capability
- Infects exclusively or preferentially tumor cells
Tumor cells
Infection
apoptosis
- Induction or stimulation of an anti-tumor
immune response.
Activation of the
immune system
Anti-tumor Virotherapy using oncolytic virus
Measles virus vaccine (MV) based anti-tumoral virotherapy
Measles virus vaccine, Schwarz strain (MV):
Dr Frédéric Tangy (Viral genomics and Vaccination laboratory, Pasteur Institut, Paris)
- Attenuated replicating vaccine strain of measles virus: Schwarz (MMR vaccine)
- Targets CD46, complement regulatory protein (wt MV targets CD150/SLAM)
- CD46 is expressed at low level by healthy cells
- CD46 is often found overexpressed on tumor cells
- Spontaneously oncolytic:
- Lymphoma, glioma, breast, ovary, prostate … (Russell SJ, Mayo Clinic, USA)
- MPM, colon and lung adenocarcinoma
Protéine L
Protéine de Fusion (F)
Hémagglutinine (H)
Protéine de Matrice (M)
Double membrane
lipidique
Phosphoprotéine (P)
ARN + Nucléoprotéine (N)
- Enveloped, non-segmented, negative-sense, single-stranded RNA (ssRNA) paramyxovirus of the genus Morbillivirus
Guillerme JB et al, Biology, 2013
- antiviral pathway defects are often found in tumor cells
Gauvrit A et al, Cancer Research, 2008; Boisgerault N et al, Biomed Res Int, 2013
Oncovirotherapy – Oncolytic properties of Measles Vaccine
Virus MV Oncolytic (Vaccine strain)
Oncolytic properties
Immunogenic apoptotic cell
death
Tumour Cell
T Lymphocytes Plasmacytoïd DC Myéloïd DC
Specific T cell
response
immunogenic molecules
DAMP & PAMP
Tumour cell infection (mesothelioma)
INFECTION
Syncitium
MV-eGFP Infected cells Mesothelioma
0 24 48 720
20
40
60
80
100
Meso13Meso47Meso56Meso96
Meso11
Time post infection (h)
% e
GFP
+ce
llsOthers
•Mesothelioma : 14/18
• Melanoma : 7/8
• Lung : 3/4
• Colorectal : 4/4
Mesothelioma (Gauvrit A et al, Cancer Research, 2008)
Lung, Colorectal adenocarcinoma (N. Boisgerault et al.,
Biomed. Res. Int., 2013)
Mesothelial cells
Normal cells: mesothelial and lung cells
Lung cells
Also for :
• Fibroblasts
• Endothelial cells
Oncovirotherapy - Live attenuated Schwarz Measles Vaccine
T Lymphocytes
Virus MV Oncolytic (Vaccine strain)
Plasmacytoïd DC
Oncolytic properties
Immunogenic apoptotic cell
death
Myéloïd DC
Specific T cell
response
Tumour Cell
immunogenic molecules
DAMP & PAMP
Danger Signals: Damage Associated Molecular Patterns
Meso13 A549 M6 HT29
100
200 ControlMV
Me
dia
n F
luo
resc
en
ce In
ten
sity
HSP70
Cross-presentation of
tumor-derived antigens
Meso13 ADK153 M6 HT29
25
50
% C
alr
eti
culi
n+ c
ell
s
Calreticulin
Phagocytosis
HMGB1 Antigen processing and
presentation
Meso13 A549 M6 HT29
100
200
300
400
HM
GB
1 (
ng/
mL)
Bianchi and Manfredi, 2007 Kepp et al., 2009
Gregoire et al, in preparation
Immune response with Live attenuated Schwarz Measles Vaccine
T Lymphocytes
Virus MV Oncolytic (Vaccine strain)
Plasmacytoïd DC
Oncolytic properties
Immunogenic apoptotic cell
death
Myéloïd DC
Specific T cell
response
Tumour Cell
immunogenic molecules
DAMP & PAMP
Activation
maturation
Plasmacytoid dendritic cells Myeloid dendritic cells
Mo-DC
pDC
MV preserves dendritic cells
(Guillerme JB et al., 2013, Clinical Cancer Research) (Gauvrit A et al., 2008, Cancer Research)
MV-infected tumor cells induced Mo-DC Maturation
HLA-A,B,C
Dci
IFN
TNF
M11
UV
M11
MV
M13
UV
M13
MV
0
1000
2000
MF
I
HLA-DR
Dci
IFN
TNF
M11
UV
M11
MV
M13
UV
M13
MV
0
50
100
MF
I
CD80
Dci
IFN
TNF
M11
UV
M11
MV
M13
UV
M13
MV
0
100
200
MF
I
CD83
Dci
IFN
TNF
M11
UV
M11
MV
M13
UV
M13
MV
0
50
100
% C
D8
3+ ce
lls
CD86
Dci
IFN
TNF
M11
UV
M11
MV
M13
UV
M13
MV
0
200
400
600
MF
I
CD40
Dci
IFN
TNF
M11
UV
M11
MV
M13
UV
M13
MV
0
200
400
600
800
1000
MF
I
DCi
IFNα
- P
oly
I:C
TN
Fα
- P
oly
I:C
Meso 11 Meso 13
MV UV MV UV
CMH I
CD86 CMH II
CD83 CD80
CD40
DCi
IFNα
- P
oly
I:C
TN
Fα
- P
oly
I:C
Meso 11 Meso 13
MV UV MV UV DCi
IFNα
- P
oly
I:C
TN
Fα
- P
oly
I:C
Meso 11 Meso 13
MV UV MV UV
(Gauvrit A et al., 2008, Cancer Research)
MUC-1 or mesothelin +Mo-DC
CD8
DCi IFN -PolyI:C UV MV0
1000
2000
3000
4000
cp
m
CD4
DCi IFN -PolyI:C UV MV0
50000
100000
cp
m
Meso 13 Meso 13
0
50
100
% C
D8+
IF
N-
+
TNFα-
PolyI:C
Meso 13 MV DCi
T CD4 Proliferation T CD8 Proliferation
Clone T CD8 anti-MUC-1 Cross-presentation (mesothelin)
(Gauvrit et al., 2008) (Gauvrit A et al., 2008, Cancer Research)
• Expression of TLR7 et TLR9 => specialized in recognition of viral nucleic acids
MV
ARNsb
IFN- Production and tumor antigen cross-presentation by pDC exposed to MV
infected tumor cells?
•Produce huge quantities of Type I IFN (- and –b) in response to virus
•Antigen cross presentation in human: •HIV (Hoeffel, G,.Immunity, 2007) (Crozat, K., J Exp Med, 2010) •Influenza (Lui, G., PLoS One, 2009)
•Antigen cross presentation in mice: •Cross-tolérance (Goubier, A., Dubois, B., Immunity, 2008) •OVA (Mouries, J., Blood, 2008)
• One of the target of imiquimod (R837, TLR7 ligand) in the treatment of basal cell carcinoma with Aldara.
Plasmacytoid dendritic cells
4°C 37°C
MV
UV
M18
MV
UV
A549
37°
C
4°C
PK
H67
BDCA-4
PK
H67
HLA-DR
pDC Mo-DC
HLA-DR Alexa568 PKH-67 MERGE
pDC +
M18 MV
pDC +
A549 MV
MV infected tumor cells are internalized by Plasmacytoid dendritic cells
CD
123
SS
C
CD86 CD40
9 %
92.4 %
20 %
90.6 %
7.42 %
CD83
92.4 %
BDCA4FSC
4,27
130,96
5,13
5,99
12,36
5,33
24
10
60
22
85
167
Meso13 MV
Meso13 UV
IL3
R848
MV
IL-3 + MV
CD83
IL3
MV
MV
+IL
3
R84
8
M1
8 M
V
M1
8 U
V
Me
so
13
MV
Me
so
13
UV
A5
49
MV
A5
49
UV
0
20
40
60
80
100
** *** **
*** ***
% P
os
itiv
e C
ell
s
CD40
IL3
MV
MV
+IL
3
R84
8
M1
8 M
V
M1
8 U
V
Me
so
13
MV
Me
so
13
UV
A5
49
MV
A5
49
UV
0
50
100
150
200
*
R-M
FI
CD86
IL3
MV
MV
+IL
3
R84
8
M1
8 M
V
M1
8 U
V
Me
so
13
MV
Me
so
13
UV
A5
49
MV
A5
49
UV
0
20
100
150
200
*
R-M
FI
MV infected tumor cells induce Plasmacytoid dendritic cells maturation
IRS661: TLR7inhibitor
IL-3 0 0,1 0,5 1 0 0,1 0,5 1 0 0,1 0,5 1
0
20
40
60
80
200
400
600
800
IL-3 + MV*10 CpG-A CpG-A
IFN
- (
ng
/ml)
IL-3 0 0,1 0,5 1 0 0,1 0,5 1 0 0,1 0,5 1
0
20
40
60
80
200
400
600
800
IL-3 + MV*10 CpG-A CpG-A
IFN
- (
ng
/ml)
IRS661
IL-3 0 0,1 0,5 1 0 0,1 0,5 1 0 0,1 0,5 1
0
20
40
60
80
200
400
600
800
IL-3 + MV*10 CpG-A CpG-A
IFN
- (
ng
/ml)
IL-3 0 0,1 0,5 1 0 0,1 0,5 1 0 0,1 0,5 1
0
20
40
60
80
200
400
600
800
IL-3 + MV*10 CpG-A M18 MV
IFN
- (
ng
/ml)
IRS661 IRS661
(TLR-9)
pD
C IF
N-α
(n
g/m
l)
IL3
MV
IL3
+ M
V
R8
48
M1
8 M
V
M1
8 U
V
A5
49
MV
A5
49
UV
0
10
20
30
200
400
600
IFN
- (
ng
/ml)
MV infected tumor cells induce production of INF type I by pDC
LT only pDC + LT
0.1µM NYESO-1[157-165]
1µM NYESO-1
[157-165]
M18
MV UV
M18 MV
A549
R848
M18 UV
10.8%
6.5%
0.8% 0.2%
0.2%
87.5%
0.1% 0.1%
0.3% 0.1%
LT - pDC
CD
8
IFN-γ
CD
8
IFN-γ
LT + pDCC
D8
IFN-γ
CD
8
IFN-γ
CD
8
IFN-γ
CD
8
IFN-γ
LT
on
ly (
n=
5)
pD
C +
LT
(n
=6
)
pD
C 0
.1 µ
M N
YE
SO
-1 (
n=
3)
pD
C +
M1
8 M
V (
n=
6)
pD
C +
M1
8 U
V(n
=5
)
0
10
20
% o
f IF
N+
T C
D8+
cell
s
MV infected tumor cells induce NY-ESO-1 cross-presentation by pDC
Cytotoxic
CD8+ T Lymphocytes (CTL)
Tumor cells
Dendritic cells
Apoptosis
MV
Gauvrit A et al, Cancer Research, 2008
Immune system activation
• Activation of myeloïd DC
• Activation of tumor antigen specific
CD8+ T lymphocytes
Dangers signal expression
• HMGB1
• Calreticuline
• HSP
• inflammasome, caspase1, IL-1b
Boisgerault N et al, in preparation
Immune system activation
• Activation of plasmacytoid DC
(production of type I interferon)
Guillerme JB et al, Clin. Cancer Res., 2013
In conclusion: Virotherapy and DC activations
IMMUNOGENIC PROPERTIES OF DEAD CELLS
CANCER RESEARCH CENTER, NANTES, FRANCE
MARC GREGOIRE
JEAN-FRANÇOIS FONTENEAU
ANNE GAUVRIT
JEAN-BAPTISTE GUILLERME
NICOLAS BOISGERAULT
VIRAL GENOMICS AND VACCINATION LABORATORY
PASTEUR INSTITUTE, PARIS, FRANCE
FRÉDÉRIC TANGY
MARIANA MESEL-LEMOINE
“DEVELOPMENT AND CLINICAL TRANSFER”
CORE FACILITY PLATFORM
DELPHINE COULAIS
CLARISSE PANTERNE