targeting checkpoint inhibitor-pd-1 for enhancing...
TRANSCRIPT
Prof. D K Mitra, Chair
Department of Transplant Immunology & Immunogenetics
All India Institute of Medical Sciences, New Delhi, India
Targeting checkpoint inhibitor-PD-1 for enhancing
efficacy of therapeutic vaccines in tuberculosis
Tuberculosis Patients
Chemotherapy Vs. Immune containment}
• Therapeutic Vaccines : Synergism
IL-1
0 (
pg
/ml)
0
50
100
150
200
IL-10
0
1
2
3
4
5
IFN-γ+ CD4+
IFN-γ+/IL-4 ratio CD4
IL-4+ CD4+
0
10
20
30
40
50
60
0 1M 3M 6M 12M 0 1M 3M 6M 12M 0 1M 3M 6M 12M
Singh et al, PlosOne’ 2013
Problem : Active suppression of T cell response
in patients
HCs
PTB Patients
0
10
20
30
40
50
60
70
80 P = 0.0001
ns
% P
osi
tive
On CD4+ FoxP-3+ cells
P = 0.00013
PD-1 PDL-1 PDL-2
Mtb. Mtb. + α-PD-1 Mtb. + α- PD (L1 / 2) Mtb. + α- PD
(1+L1 / 2)
CD3
IFN
-γ
Media
10 0 10 1 10 2 10 3 10 4
10 0
10 1
10 2
10 3
10 4
4.14%
10 0 10 1 10 2 10 3 10 4
10 0
10 1
10 2
10 3
10 4
8.49%
100 10 1 10 2 10 3 10 4
10 0
10 1
10 2
10 3
10 4
7.26%
10 0 10 1 10 2 10 3 10 4
10 0
10 1
10 2
10 3
10 4
9.7%
10 0 10 1 10 2 10 3 10 4
10 0
10 1
10 2
10 3
10 4
1.67%
Media Mtb.Ag Mtb.Ag+α PD-1 Mtb.Ag+ α PD-L1
PBL
BAL
MTB10 0 10 1 10 2 10 3 10 4
10 0
10 1
10 2
10 3
10 4
10 0 10 1 10 2 10 3 10 4
10 0
10 1
10 2
10 3
10 4
10 0 10 1 10 2 10 3 10 4
10 0
10 1
10 2
10 3
10 4
0.62%
0.32% 2.46%
10 0 10 1 10 2 10 3 10 4
10 0
10 1
10 2
10 3
10 4
10 0 10 1 10 2 10 3 10 4
10 0
10 1
10 2
10 3
10 4
10 0 10 1 10 2 10 3 10 4
100
101
102
103
104
10 0 10 1 10 2 10 3 104
10 0
10 1
10 2
10 3
10 4
4.2% 6.65%
14.4% 15.7%
10 0 10 1 10 2 10 3 10 410 0
10 1
10 2
10 3
10 4
6.16%
CD3
IFN
-γImpact of PD-1 on Mtb. sepcific Teff cells is
Pronounced in lungs of Miliary Tuberculosis
% I
FN
-Υ+
T c
ells
0 1 2 3 40.0
2.5
5.0
7.5
10.0
12.5
15.0
TB-PE-PBL
TB-PE-PE
MTB-PBL
MTB-BAL*
** **
Singh et al, Clin. & Exptl. Immunol.’ 2016
❖ Loss of poly-functional T cell in TB patients
Latent TB (PPD+)
Active TB
Household Contacts (PPD-)
%
Po
siti
ve
CD
4 T
cel
ls
*
*
*
*
(N=15)
Giuseppe Pantaleo et al , Nature Medicine, 2011
✓Dominant TNF-α+ Mycobacterium tuberculosis–specific CD4+ T cell
responses discriminate between latent infection and active disease
Preferential rescue of IFN-γ+ TNF-α+
producers after PD-1 blocking.
TNF-α
IFN-γ
Media Mtb. Ag Mtb. Ag + a-PD-1
100
101
102
103
104
100
101
102
103
104
TB-148,M,1.003ÉCD4
FL2-H: TNFa PE
FL
1-H
: IF
Ng
FIT
C
0.98 0.65
0.4997.9
100
101
102
103
104
100
101
102
103
104
TB-148,Ag,S1.002ÉCD4
FL2-H: TNFa PE
FL
1-H
: IF
Ng
FIT
C
2.05 1.07
2.9393.9
100
101
102
103
104
100
101
102
103
104
TB-148,Ag+aPD1,2.006ÉCD4
FL2-H: TNFa PE
FL
1-H
: IF
Ng
FIT
C
6.29 4.08
3.9185.7
0.65%0.98%
0.49%
2.05% 1.07%
2.93%
4.29% 4.08%
3.91%
IFN6%
TNF4% IL-2
4%
IFN/TNF40%IFN/1L-2
12%
TNF/IL-218%
TP16%
N=5
IFN-γ TNF-α+ IFN-γ+TNF-α+
*
*%
PD
-1 E
XP
RE
SS
ION
TNF-α milieu promotes Necrosis
whereas IFN-γ alone and in
combination with TNF-α results
in apoptotic death of M.tb.
infected MDMs
Recombinant IFN-γ
Recombinant TNF-α
Recombinant IFN-γ +
TNF-α
10%
33%57%
2%
12%
86%
Early Apoptotic
Late Apoptotic
Necrotic
8%
62%
30%
(N=3)
Cytokine Milieu influences the fate of Mtb infected MDMs
*
Mtb (H37Rv) + + + +
rIFN-γ - - + -
rTNF-α - + - -
rIFN-γ+rTNF-α - - + +
Col
ony-
form
ing
Uni
ts
Tregs + - + +
αPD-1 - - - +
Tregs + - + +
αPD-1 - - - +
Tregs + - + +
* *
αPD-1 - - - +
IFN-γ + cells TNF-α + cells IFN-γ +TNF-α + cells P
erce
nta
ge
Incr
ease
** *
H37R
v
%
Nec
rosi
s o
f
CD
14
+
cell
s
H37Rv+anti PD-1
*
% A
po
pto
sis
of
CD
14
+
cell
s
H37Rv H37Rv+anti
PD-1
(N=7)
*
**
*
** **C
FU
/ml
Mtb (H37Rv) + + + + +
α PD-1 - + + + +
α IFN-γ - - + - +
αTNF-α - - - + +
Naïve a-PD1 ATTIsotype
Control
ATT + a-PD1
Administration of anti-PD-1 reduces CFU burden in lungs and spleen of miceMtb infected mice
40X
0
0.05
0.1
0.15
0.2
0.25
0.3
0
0.05
0.1
0.15
Lungs
Spleen
*
*
**
***
**
CD
4+IF
N-γ
+T
NF
-α+
(%
)
Lu
ng
s (l
og
10C
FU
/g)
*
*
0
1
2
3
4
5
6
Sp
leen
(lo
g10C
FU
/g)
*
**
*
***
CD
4+IF
N-γ
+T
NF
-α+(%
)
Mtb specific Poly-functional T cellsCFU
Naive α PD-1 ATT+ α PD-1ATT
Naive α PD-1 ATT+ α PD-1ATT
Isotype
Controlα PD-1 ATT+ α PD-1ATT
Isotype
Controlα PD-1 ATT+ α PD-1ATT
5
5.5
6
6.5
7
7.5 *
ID93 antigen along with α-PD-1 enhances the bacterial
clearance in in vitro MDM model C
FU
/ml (1
04)
Pe
rce
nt
red
uc
tio
n in
CF
U
Gene Efflux Pump Drugs
RV1258
Major Facilitator Superfamily
(MFS)
Rifampicin, Fluoroquinolones
Mmp17
Resistance-nodulation-division
(RND) family
Isoniazid
Rv1258 Mmp17
The impact of cytokines on the efflux pumps of M. tuberculosis
➢ Inhibiting PD1
Rescues protective T cells
Synargism with chemotherapy
Better bacillary clearance
Prevention of relapse
➢Likely to potentiate the effect of therapeutic vaccines
➢May aid to therapeutic vaccination in MDR Tuberculosis
Biased Imagination
Prabhat Sharma
Funding : DBT, ICMR, Govt of India
Immunology, AIIMS, New Delhi
AIIMS, New Delhi
Medicine : A B Dey Steve Reed, Rhea ColerIDRI, Seattle, USA
Pulmonary Medicine R Guleria, Anant Mohan
Microbiology, V P Chest
Institute, New Delhi
Mandira V Basil
Thanks for patient hearing
U D Gupta, Pushpa GuptaJALMA, ICMR, Agra
Divya Kamboj Amar Singh