A n t i - T I G I T -...
1
Anti-TIGIT facilitates potent anti-tumor immune response in multiple mouse tumor models. Anti-TIGIT potentiates tumor antigen-specific T-cell IFN-g production and increases T cell cytotoxicity against parental tumor. Anti-TIGIT combines with anti-PDL1/anti-PD1 to enhance efficacy as compared to single agents in CT26.WT tumor models. Combination of anti-TIGIT and anti-PDL1 increases tumor regression, increases tumor-specific T cells and generates long- term immune memory against the originally implanted tumor cells. In Vivo Studies: The murine colon carcinoma (CT26-WT, ATCC CRL-2638) was obtained from American Type Culture Collection. Single cell suspensions of CT26.WT or MC38 tumor cells were injected subcutaneously into the flanks of 7-8 week old BALB/c or C57BL/6N mice, respectively. One week following tumor implantation, mice were randomized by tumor volume and injected i.p. with anti-TIGIT and anti- PDL1/anti-PD1 antibodies twice a week for 3 weeks (6 doses). Isotype antibodies were used for control. Tumor volumes were monitored by measuring two bisecting diameters of each tumor with electronic calipers. Tumor volumes were calculated using the formula: V=0.5ab2, with a as the larger diameter and b as the smaller diameter. ELISPOT: Splenocytes were cultured in the presence or absence of tumor specific CD8 T cell peptide in T cell media for 48 hrs. followed by the ELISPOT assay as described by manufacturer’s instruction. Flow Cytometry: Single cell suspensions of splenocytes or tumor digests were pretreated with Fc block and stained with indicated antibodies and their isotype controls followed by fixing. Cells were analyzed by flow cytometry (FACS Canto II) and data was processed using Diva software. Tumor specific T cells were analyzed in the periphery by using AH1 dextramer from Immudex. T cell cytotoxicity assay: Ten million splenocytes were cultured with a tumor-specific CD8+ T cell peptide for 7 days, washed and counted. These effector T cells were co-cultured with calcein AM labeled CT26.WT target tumor cells (E:T of 25:1 & 50:1) for four hours in triplicate wells in a 96-well plate and the supernatants were collected and measured for the release of calcein from tumor cells. TIGIT (T cell immunoreceptor with Ig and ITIM domains) has been recently described as an inhibitory receptor which blocks CD8 T cell-mediated anti-tumor immune responses. We have generated an anti- mouse TIGIT antibody (313R12) to evaluate drug efficacy and mechanism of action in pre-clinical tumor models. Anti-TIGIT as a single agent promoted an anti-tumor immune response in multiple syngeneic mouse tumor models. Anti-TIGIT enhanced tumor specific T cell responses, particularly of the Th1 type and reduced Th2 type responses and also increased the function of cytotoxic T cells. Furthermore, anti-TIGIT displayed combination activity with immune checkpoint inhibitors anti-PD1 and anti-PDL1 in inhibiting tumor growth, promoting complete tumor rejection and significantly increasing mouse survival in the murine CT26 colon carcinoma model as compared to controls and single agents alone. Mice “cured” with anti-TIGIT/anti-PDL1 or anti-TIGIT/anti-PD1 combination treatments were protected from subsequent re- challenges with increasing numbers of tumor cells, suggesting the existence of immunologic memory. IL2 and tumor-specific IFN-g production by splenic T cells were increased in the rescued mice in combination treatment compared to controls. Additionally, both effector and memory CD8+ T cell frequencies were increased within the total CD8+ T cell population in the rescued mice. We also demonstrated an increase in tumor-specific CD8 T cells in the periphery in anti-TIGIT/anti-PDL1 combination treatment compared to controls. Therefore, these results show that co-targeting of TIGIT and PD1 or PDL1 may be an effective and durable cancer therapy by increasing T cell-mediated anti-tumor immune responses and promoting long- term immunological memory. Anti-TIGIT induces T cell-mediated anti-tumor immune responses and combines with immune checkpoint inhibitors to enhance strong and long term anti-tumor immunity Minu K. Srivastava, Rui Yun, Hyun-Bae Jie, Erin Mayes, Janice Yu, Fumiko Axelrod, Ming-Hong Xie, Jorge Monteon, Andrew Lam, May Ji, Yuwang Liu, John Lewicki, Tim Hoey, Austin Gurney, and Angie Inkyung Park OncoMed Pharmaceuticals, Inc., Redwood City, CA MATERIALS AND METHODS ABSTRACT RESULTS AND CONCLUSIONS Modes of action of TIGIT Anti - TIGIT reduces tumor growth in CT26.WT and MC38 murine CRC models Anti - TIGIT increased tumor specific CD8+ T cell - mediated IFN - γ production and T cell cytotoxicity against parental tumor cells Fig 3. CT26.WT tumor bearing BALB/c mice were dosed with control or anti-TIGIT antibody. At the end of experiment, spleen cells from each group were isolated and tested for functions. (A-B) IFN-g ELISPOT. (C) Tumor-specific T cell cytotoxicity Combination of anti - TIGIT and anti - PD1 inhibits tumor growth and increases survival in CT26.WT tumor model Fig 4. The combination of anti-TIGIT and anti-PD1 resulted in significant tumor growth inhibition and tumor regression. (A) CT26.WT tumor bearing BALB/c mice were dosed with control, anti-PDL1, anti-TIGIT, or combination of both antibodies twice a week for 3 weeks. (A) Tumor growth curves (n=15 mice per group). (B) Survival curve. ** indicate P < 0.0001.* indicate P < 0.05 against the control group. Anti - TIGIT + anti - PD1 facilitates long - term immune memory against parental tumor cells Anti - TIGIT + anti - PDL1 facilitates long - term immune memory against parental tumor cells SUMMARY Fig 1: TIGIT can block T cell lymphocytes through multiple mechanisms of action. APC, antigen-presenting cell, PVR, poliovirus receptor, Tigit, T cell immunoreceptor and ITIM domain; Teff, effector T cell; Treg, regulatory T cell. Manieri et al, Trends in Immunology. 2016 A 0 10 20 30 0 500 1000 1500 Days Tumor Volume (mm 3 ) Control Below 250mm 3 : 1/20 0 10 20 30 0 500 1000 1500 Days Tumor Volume (mm 3 ) Anti-TIGIT Below 250mm 3 : 13/20 CT26WT Tumor model 0 5 10 15 20 25 0 500 1000 1500 Days Tumor Volume (mm 3 ) Control Below 250mm 3 : 1/10 0 5 10 15 20 25 0 500 1000 1500 Days Tumor Volume (mm 3 ) Anti-TIGIT Below 250mm 3 : 4/10 MC38 Tumor model B Fig 2. Anti-TIGIT reduces tumor growth in CT26.WT & MC38 mouse tumor models. BALB/c or C57BL/6N mice were inoculated with 3x10 4 CT26.WT cells (A) or 4x10 5 MC38 cells (B), and at day 7 mice were treated with isotype control or anti-TIGIT antibody. Spaghetti plots show individual mice tumor growth over time. Control Anti-TIGIT -Ve +Ve A 0 200 400 600 800 1,000 1,200 Control Anti-TIGIT TOD (x1000) IFNg TOD No Pepide peptide * B 0 10 20 30 40 50 25:1 50:1 % Specific Lysis T cell cytotoxicity against CT26 Control 313R12 C 0 10 20 30 40 50 0 1000 2000 3000 Days Tumor Volume (mm 3 ) Control 0 10 20 30 40 50 0 1000 2000 3000 Days Tumor Volume (mm 3 ) Anti-TIGIT 0 10 20 30 40 50 0 1000 2000 3000 Days Tumor Volume (mm 3 ) Anti-PD1 0 10 20 30 40 50 0 1000 2000 3000 Days Tumor Volume (mm 3 ) Combo A 0 50 100 150 200 0 50 100 Days Post Treatment Percent survival Control Anti- PD1 Anti-TIGIT Combo ** * * B CT26.WT 30K 1 st re-challenge (1X) 2nd re-challenge (5X) Day 128 from 1 st tumor injection and 100 days from last dose Day 0 Day 255 from 1st tumor injection and 227 days from last dose 3rd re-challenge (10X) Day 325 from 1 st tumor injection and 297 days from last dose Control Anti-TIGIT Anti-PDL1 Combo 0 1000 2000 3000 Tumor Volume (mm 3 ) Control Anti-TIGIT Anti-PDL1 Combo 0 1000 2000 3000 Control Anti-TIGIT Anti-PDL1 Combo 0 1000 2000 3000 A B C Tumor Free Mice (Re-challenge study) Treatment Initial 1st re challenge 2nd re challenge 3rd re challenge 30k 30k 150k 300k Control 0% (0/10) 0% (0/10) 0% (0/10) Anti-TIGIT 65% (13/20) 84% (11/13) 100% (11/11) 100% (11/11) Anti-PDL1 20% (2/10) 100% (2/2) 100% (2/2) 100% (2/2) Combo 70% (7/10) 100% (7/7) 100% (7/7) 100% (7/7) Fig 7 : The combination of anti-TIGIT and anti-PDL1 resulted in long-term protection against the original tumor. Mice “cured” of CT26.WT tumors were subjected to a series of tumor re-challenges. 1 st re-challenge using 30K tumor cells, (B) 2 nd re-challenge using 150K cells (C)3 rd re- challenge using 300K cells. Naïve mice were also inoculated with the same number of cells for controls. CT26.WT 30K 1 st re-challenge (2X) 2nd re-challenge (5X) Day 168 from 1 st tumor injection and 144 days from last dose Day 0 Day 316 from 1st tumor injection and 292 days from last dose Tumor Free Mice (Re-challenge study) Treatment Initial 1st re-challenge 2nd re-challenge 30k 60k 150k Control 0% (0/10) 0% (0/10) Anti-TIGIT 20% (3/15) 100% (2/2) 100% (2/2) Anti-PD1 40% (6/15) 100% (6/6) 100% (6/6) Combo 86% (13/15) 100% (13/13) 100% (12/12) Fig 5 : The combination of anti-TIGIT and anti-PD1 resulted in a long-term protection against the original tumor. Mice “cured” of CT26.WT colon cancer were re-challenged in the opposite flank of the original injection site with the indicated number of tumor cells (1 st : 60K, 2 nd : 150K CT26.WT cells). Naïve mice were also inoculated with the same number of cells for controls. Spleens of the “cured” mice after 2nd re-challenge were analyzed for IFNg and IL2 by ELISPOT. NTB: Non tumor bearing mice. . Control Anti-PD1 Anti-TIGIT Combo NTB 0 200000 400000 600000 IL-2 TOD IL-2 Control Anti-PD1 Anti-TIGIT Combo NTB 0 100000 200000 300000 IFN- g TOD IFN- g Combination of anti - TIGIT and anti - PDL1 inhibits tumor growth and increases survival in CT26.WT tumor model 0 20 40 60 80 -500 0 500 1000 1500 2000 2500 3000 3500 Days Tumor Volume (mm 3) 2/10 CR Control 0 20 40 60 80 -500 0 500 1000 1500 2000 2500 3000 3500 4/10 Days Tumor Volume (mm 3 ) Anti-TIGIT 0 20 40 60 80 -500 0 500 1000 1500 2000 2500 3000 3500 Days Tumor Volume (mm 3 ) 2/9 CR Anti-PDL1 0 20 40 60 80 -500 0 500 1000 1500 2000 2500 3000 3500 7/9 CR Days Tumor Volume (mm 3 ) Combo 0 20 40 60 80 0 20 40 60 80 100 Control Anti-TIGIT Anti-PDL1 Combo Days Post Treatment Percent survival ** * Fig 6. The combination of anti- TIGIT and anti-PDL1 resulted in significant tumor growth inhibition and tumor regression. CT26.WT tumor bearing BALB/c mice were treated i.p. with control, anti-PDL1, anti-TIGIT, or combination twice a week for 3 weeks. (A) Individual mice tumor growth curves (n=9 mice per group). (B) Survival curve. ** indicate SEM P < 0.009.* indicate SEM P < 0.05 against the control group. Combination of anti - TIGIT and anti - PDL1 increases strong anti - tumor immune responses in re - challenged mice Control Combo IL2 Control Combo IFNg -Ve +Ve -Ve +Ve A B 0 5 10 15 20 25 Control Combo % Gated on CD8T CT26 CD8_EM * 0 2 4 6 8 Control Combo % Gated on SSC CT26_CD8 * Control Combo C Control Neg. dextramer Combo AH1 dextramer D Fig 8 : Spleens of the “cured” mice after 3 rd re-challenge from Fig. 7C were analyzed for IL2 (A) and IFNg (B) production by ELISPOT and T cell memory population (C) and tumor-specific (AH1+) T cells (D). A B
Transcript of A n t i - T I G I T -...
Anti-TIGIT facilitates potent anti-tumor immune response in multiple mouse tumor models.
Anti-TIGIT potentiates tumor antigen-specific T-cell IFN-g production and increases T cell cytotoxicity against parental tumor.
Anti-TIGIT combines with anti-PDL1/anti-PD1 to enhance efficacy as compared to single agents in CT26.WT tumor models.
Combination of anti-TIGIT and anti-PDL1 increases tumor regression, increases tumor-specific T cells and generates long-
term immune memory against the originally implanted tumor cells.
In Vivo Studies: The murine colon carcinoma (CT26-WT, ATCC CRL-2638) was obtained from American
Type Culture Collection. Single cell suspensions of CT26.WT or MC38 tumor cells were injected
subcutaneously into the flanks of 7-8 week old BALB/c or C57BL/6N mice, respectively. One week following
tumor implantation, mice were randomized by tumor volume and injected i.p. with anti-TIGIT and anti-
PDL1/anti-PD1 antibodies twice a week for 3 weeks (6 doses). Isotype antibodies were used for control.
Tumor volumes were monitored by measuring two bisecting diameters of each tumor with electronic
calipers. Tumor volumes were calculated using the formula: V=0.5ab2, with a as the larger diameter and b
as the smaller diameter.
ELISPOT: Splenocytes were cultured in the presence or absence of tumor specific CD8 T cell peptide in T
cell media for 48 hrs. followed by the ELISPOT assay as described by manufacturer’s instruction.
Flow Cytometry: Single cell suspensions of splenocytes or tumor digests were pretreated with Fc block
and stained with indicated antibodies and their isotype controls followed by fixing. Cells were analyzed by
flow cytometry (FACS Canto II) and data was processed using Diva software. Tumor specific T cells were
analyzed in the periphery by using AH1 dextramer from Immudex.
T cell cytotoxicity assay: Ten million splenocytes were cultured with a tumor-specific CD8+ T cell peptide
for 7 days, washed and counted. These effector T cells were co-cultured with calcein AM labeled CT26.WT
target tumor cells (E:T of 25:1 & 50:1) for four hours in triplicate wells in a 96-well plate and the
supernatants were collected and measured for the release of calcein from tumor cells.
TIGIT (T cell immunoreceptor with Ig and ITIM domains) has been recently described as an inhibitory
receptor which blocks CD8 T cell-mediated anti-tumor immune responses. We have generated an anti-
mouse TIGIT antibody (313R12) to evaluate drug efficacy and mechanism of action in pre-clinical tumor
models. Anti-TIGIT as a single agent promoted an anti-tumor immune response in multiple syngeneic
mouse tumor models. Anti-TIGIT enhanced tumor specific T cell responses, particularly of the Th1 type and
reduced Th2 type responses and also increased the function of cytotoxic T cells. Furthermore, anti-TIGIT
displayed combination activity with immune checkpoint inhibitors anti-PD1 and anti-PDL1 in inhibiting
tumor growth, promoting complete tumor rejection and significantly increasing mouse survival in the
murine CT26 colon carcinoma model as compared to controls and single agents alone. Mice “cured” with
anti-TIGIT/anti-PDL1 or anti-TIGIT/anti-PD1 combination treatments were protected from subsequent re-
challenges with increasing numbers of tumor cells, suggesting the existence of immunologic memory. IL2
and tumor-specific IFN-g production by splenic T cells were increased in the rescued mice in combination
treatment compared to controls. Additionally, both effector and memory CD8+ T cell frequencies were
increased within the total CD8+ T cell population in the rescued mice. We also demonstrated an increase in
tumor-specific CD8 T cells in the periphery in anti-TIGIT/anti-PDL1 combination treatment compared to
controls. Therefore, these results show that co-targeting of TIGIT and PD1 or PDL1 may be an effective and
durable cancer therapy by increasing T cell-mediated anti-tumor immune responses and promoting long-
term immunological memory.
Anti-TIGIT induces T cell-mediated anti-tumor immune responses and combines with immune checkpoint
inhibitors to enhance strong and long term anti-tumor immunity Minu K. Srivastava, Rui Yun, Hyun-Bae Jie, Erin Mayes, Janice Yu, Fumiko Axelrod, Ming-Hong Xie, Jorge Monteon, Andrew Lam, May Ji, Yuwang Liu, John
Lewicki, Tim Hoey, Austin Gurney, and Angie Inkyung Park OncoMed Pharmaceuticals, Inc., Redwood City, CA
MATERIALS AND METHODS
ABSTRACT
RESULTS AND CONCLUSIONS
Modes of action of TIGIT
Anti-TIGIT reduces tumor growth in CT26.WT and MC38 murine CRC
models
Anti-TIGIT increased tumor specific CD8+ T cell-mediated IFN-γ
production and T cell cytotoxicity against parental tumor cells
Fig 3. CT26.WT tumor
bearing BALB/c mice
were dosed with
control or anti-TIGIT
antibody. At the end of
experiment, spleen
cells from each group
were isolated and
tested for functions.
(A-B) IFN-g ELISPOT.
(C) Tumor-specific T
cell cytotoxicity
Combination of anti-TIGIT and anti-PD1 inhibits tumor growth and
increases survival in CT26.WT tumor model
Fig 4. The combination of anti-TIGIT and anti-PD1
resulted in significant tumor growth inhibition and tumor
regression. (A) CT26.WT tumor bearing BALB/c mice
were dosed with control, anti-PDL1, anti-TIGIT, or
combination of both antibodies twice a week for 3
weeks. (A) Tumor growth curves (n=15 mice per group).
(B) Survival curve. ** indicate P < 0.0001.* indicate P <
0.05 against the control group.
Anti-TIGIT + anti-PD1 facilitates long-term immune memory against
parental tumor cells
Anti-TIGIT + anti-PDL1 facilitates long-term immune memory against
parental tumor cells
SUMMARY
Fig 1: TIGIT can block T cell
lymphocytes through multiple
mechanisms of action. APC,
antigen-presenting cell, PVR,
poliovirus receptor, Tigit, T cell
immunoreceptor and ITIM
domain; Teff, effector T cell; Treg,
regulatory T cell.
Manieri et al, Trends in Immunology. 2016
A
0 1 0 2 0 3 0
0
5 0 0
1 0 0 0
1 5 0 0
D a y s
Tu
mo
r V
olu
me
(m
m3
)
C o n tro l
B e lo w 2 5 0 m m3: 1 /20
0 1 0 2 0 3 0
0
5 0 0
1 0 0 0
1 5 0 0
D a y s
Tu
mo
r V
olu
me
(m
m3
)
A n ti-T IG IT
B e lo w 2 5 0 m m3: 1 3 /20
C T 2 6 W T T u m o r m o d e l
0 5 1 0 1 5 2 0 2 5
0
5 0 0
1 0 0 0
1 5 0 0
D a y s
Tu
mo
r V
olu
me
(m
m3
)
C o n tro l
B e lo w 2 5 0 m m3: 1 /10
0 5 1 0 1 5 2 0 2 5
0
5 0 0
1 0 0 0
1 5 0 0
D a y s
Tu
mo
r V
olu
me
(m
m3
)
A n ti-T IG IT
B e lo w 2 5 0 m m3: 4 /10
M C 3 8 T u m o r m o d e lB
Fig 2. Anti-TIGIT reduces tumor growth in CT26.WT & MC38 mouse
tumor models. BALB/c or C57BL/6N mice were inoculated with 3x104
CT26.WT cells (A) or 4x105 MC38 cells (B), and at day 7 mice were
treated with isotype control or anti-TIGIT antibody. Spaghetti plots show
individual mice tumor growth over time.
Control Anti-TIGIT
-Ve +Ve
A
0
200
400
600
800
1,000
1,200
Control Anti-TIGIT
TO
D (
x1000)
IFNg TOD
No Pepide
peptide
* B
0
10
20
30
40
50
25:1 50:1
% S
pe
cif
ic L
ysis
T cell cytotoxicity against CT26
Control 313R12
C
0 1 0 2 0 3 0 4 0 5 0
0
1 0 0 0
2 0 0 0
3 0 0 0
D a y s
Tu
mo
r V
olu
me
(m
m3
)
C o n tro l
0 1 0 2 0 3 0 4 0 5 0
0
1 0 0 0
2 0 0 0
3 0 0 0
D a y sT
um
or V
olu
me
(m
m3
)
A n ti-T IG IT
0 1 0 2 0 3 0 4 0 5 0
0
1 0 0 0
2 0 0 0
3 0 0 0
D a y s
Tu
mo
r V
olu
me
(m
m3
)
A n ti-P D 1
0 1 0 2 0 3 0 4 0 5 0
0
1 0 0 0
2 0 0 0
3 0 0 0
D a y s
Tu
mo
r V
olu
me
(m
m3
)
C o m b o
A
0 5 0 1 0 0 1 5 0 2 0 0
0
5 0
1 0 0
D a y s P o s t T re a tm e n t
Pe
rc
en
t s
urv
iva
l
C o n tro l
A n ti- P D 1
A n ti-T IG IT
C o m b o
* *
*
*
B
CT26.WT 30K 1st re-challenge (1X) 2nd re-challenge (5X)
Day 128 from 1st tumor
injection and 100 days
from last dose
Day 0 Day 255 from 1st tumor
injection and 227 days
from last dose
3rd re-challenge (10X)
Day 325 from 1st tumor
injection and 297 days
from last dose
Contr
ol
Anti-T
IGIT
Anti-P
DL1
Com
bo
0
1 0 0 0
2 0 0 0
3 0 0 0
Tu
mo
r V
olu
me
(m
m3
)
Co
ntr
ol
An
ti-T
IGIT
An
ti-P
DL
1
Co
mb
o
0
1 0 0 0
2 0 0 0
3 0 0 0
Co
ntr
ol
An
ti-T
IGIT
An
ti-P
DL
1
Co
mb
o
0
1 0 0 0
2 0 0 0
3 0 0 0A B C
Tumor Free Mice (Re-challenge study)
Treatment Initial 1st re
challenge 2nd re
challenge 3rd re
challenge 30k 30k 150k 300k
Control 0% (0/10) 0% (0/10) 0% (0/10)
Anti-TIGIT 65%
(13/20) 84% (11/13) 100% (11/11) 100% (11/11)
Anti-PDL1 20%
(2/10) 100% (2/2) 100% (2/2) 100% (2/2)
Combo 70%
(7/10) 100% (7/7) 100% (7/7) 100% (7/7)
Fig 7 : The combination of anti-TIGIT and anti-PDL1 resulted in long-term protection against the original tumor. Mice “cured” of CT26.WT tumors
were subjected to a series of tumor re-challenges. 1st re-challenge using 30K tumor cells, (B) 2nd re-challenge using 150K cells (C)3rd re-
challenge using 300K cells. Naïve mice were also inoculated with the same number of cells for controls.
CT26.WT 30K 1st re-challenge (2X) 2nd re-challenge (5X)
Day 168 from 1st tumor injection
and 144 days from last dose Day 0 Day 316 from 1st tumor injection and
292 days from last dose
Tumor Free Mice (Re-challenge study)
Treatment Initial 1st re-challenge 2nd re-challenge 30k 60k 150k
Control 0% (0/10) 0% (0/10) Anti-TIGIT 20% (3/15) 100% (2/2) 100% (2/2) Anti-PD1 40% (6/15) 100% (6/6) 100% (6/6) Combo 86% (13/15) 100% (13/13) 100% (12/12)
Fig 5 : The combination of anti-TIGIT and anti-PD1 resulted in a
long-term protection against the original tumor. Mice “cured” of
CT26.WT colon cancer were re-challenged in the opposite
flank of the original injection site with the indicated number of
tumor cells (1st: 60K, 2nd: 150K CT26.WT cells). Naïve mice
were also inoculated with the same number of cells for controls.
Spleens of the “cured” mice after 2nd re-challenge were
analyzed for IFNg and IL2 by ELISPOT. NTB: Non tumor
bearing mice.
.
Co
ntr
ol
An
ti-P
D1
An
ti-T
IGIT
Co
mb
o
NT
B
0
2 0 0 0 0 0
4 0 0 0 0 0
6 0 0 0 0 0
IL-2
TO
D
IL -2
Co
ntr
ol
An
ti-P
D1
An
ti-T
IGIT
Co
mb
o
NT
B
0
1 0 0 0 0 0
2 0 0 0 0 0
3 0 0 0 0 0
IFN
-g T
OD
IF N -g
Combination of anti-TIGIT and anti-PDL1 inhibits tumor growth and
increases survival in CT26.WT tumor model
0 2 0 4 0 6 0 8 0
-5 0 0
0
5 0 0
1 0 0 0
1 5 0 0
2 0 0 0
2 5 0 0
3 0 0 0
3 5 0 0
D a y s
Tu
mo
r V
olu
me
(m
m3
)
2 /1 0 C R
C o n tro l
0 2 0 4 0 6 0 8 0
-5 0 0
0
5 0 0
1 0 0 0
1 5 0 0
2 0 0 0
2 5 0 0
3 0 0 0
3 5 0 0
4 /1 0
D a y s
Tu
mo
r V
olu
me
(m
m3
)
A n ti-T IG IT
0 2 0 4 0 6 0 8 0
-5 0 0
0
5 0 0
1 0 0 0
1 5 0 0
2 0 0 0
2 5 0 0
3 0 0 0
3 5 0 0
D a y s
Tu
mo
r V
olu
me
(m
m3
)
2 /9 C R
A n ti-P D L 1
0 2 0 4 0 6 0 8 0
-5 0 0
0
5 0 0
1 0 0 0
1 5 0 0
2 0 0 0
2 5 0 0
3 0 0 0
3 5 0 0
7 /9 C R
D a y s
Tu
mo
r V
olu
me
(m
m3
)
C o m b o
0 2 0 4 0 6 0 8 0
0
2 0
4 0
6 0
8 0
1 0 0
C o n t r o l
A n t i - T IG IT
A n t i -P D L 1
C o m b o
D a y s P o s t T re a tm e n t
Pe
rc
en
t s
urv
iva
l * *
*
Fig 6. The combination of anti-
TIGIT and anti-PDL1 resulted in
significant tumor growth inhibition
and tumor regression. CT26.WT
tumor bearing BALB/c mice were
treated i.p. with control, anti-PDL1,
anti-TIGIT, or combination twice a
week for 3 weeks. (A) Individual
mice tumor growth curves (n=9
mice per group). (B) Survival curve.
** indicate SEM P < 0.009.* indicate
SEM P < 0.05 against the control
group.
Combination of anti-TIGIT and anti-PDL1 increases strong anti-tumor
immune responses in re-challenged mice
Control Combo
IL2
Control Combo
IFNg
-Ve +Ve -Ve +Ve
A B
0
5
10
15
20
25
Control Combo
% G
ate
d o
n C
D8T
CT26 CD8_EM *
0
2
4
6
8
Control Combo
% G
ate
d o
n S
SC
CT26_CD8
*
Control Combo C
Control Neg. dextramer Combo
AH1 dextramer D
Fig 8 : Spleens of the “cured” mice after 3rd re-challenge
from Fig. 7C were analyzed for IL2 (A) and IFNg (B)
production by ELISPOT and T cell memory population (C)
and tumor-specific (AH1+) T cells (D).
A B
![Adapters in lymphocyte signaling · immunoreceptor (e.g., the T cell receptor [TCR], the B cell receptor [BCR], or the Fc receptors [FcRs]), and a costim-ulatory signal (see Frauwirth](https://static.fdocuments.in/doc/165x107/5c5fb15d09d3f2a3398b4758/adapters-in-lymphocyte-signaling-immunoreceptor-eg-the-t-cell-receptor-tcr.jpg)
