Adnan Deronic*, Anneli Nilsson, Mia Thagesson, Doreen ...Adnan Deronic*, Anneli Nilsson, Mia...
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Adnan Deronic*, Anneli Nilsson, Mia Thagesson, Doreen Werchau, Niina Veitonmäki, Per Norlén, Christina Furebring and Peter Ellmark Alligator Bioscience AB, Lund, Sweden, *Presenting author ADC-1013 (JNJ-64457107) is a human CD40 agonistic IgG1 antibody. It activates antigen-presenting cells (APC) by stimulating CD40 on the surface of e.g. dendritic cells. Mediated by its effect on APC, ADC-1013 treatment results in activation of tumor-directed cytotoxic T cells with capacity to eradicate tumors. Mechanism of action of ADC-1013 ADC-1013 eradicates the syngeneic MB49 bladder tumor in hCD40tg mice OVA vaccination combined with ADC-1013 treatment delays EG7.OVA tumor growth Tumor volume (mm 3 ) Figure 6 – Effect of OVA vaccination combined with prophylactic ADC-1013 treatment on EG7.OVA tumor growth. Mice (hCD40tg) were administered 200 μg OVA i.v. and 100 μg ADC-1013 i.p. as shown in Figure 3 and were inoculated with 1x10 6 EG7.OVA cells s.c. No additional treatments were administered following tumor inoculation. Graphs show tumor volume ± SEM (left) and percent survival (right) from two pooled experiments. OVA vaccination model for evaluation of ADC-1013 effect on antigen-specific T cells Figure 3 – Experimental strategy for the OVA vaccination model. Mice (hCD40tg) were divided in five groups. Three groups were administered 200 μg OVA i.v. and the remaining two received PBS i.v. on days 0 and 7. Mice were additionally treated with 100 μg ADC-1013 (white arrows) every 2-3 days or every 7 days as outlined. Ctr IgG (100 μg, grey stippled arrows) was administered to two of the groups as control. On day 14, a cohort of mice from each group was sacrificed for FACS analysis of spleens. The remaining mice were inoculated with 1x10 6 EG7.OVA cells s.c. and tumor growth evaluated. ADC-1013 every 2-3 days Ctr IgG ADC-1013 every 2-3 days 0 2 5 7 9 12 14 EG7.OVA s.c. Day: PBS/OVA i.v. Ctr IgG PBS + OVA + ADC-1013 every 7 days Tumor growth FACS PBS/OVA i.v. Summary IgG1 Fc CD40-binding domain Aim: To demonstrate the in vivo effect of systemically administered ADC-1013 on APC and antigen-specific T cells. Methods: Mice transgenic for human CD40 (hCD40tg) were immunized with ovalbumin (OVA) and treated systemically with ADC-1013 and APC and T cell activation analyzed by flow cytometry. Mice were also inoculated with an OVA-expressing tumor and anti- tumor efficacy of this treatment evaluated. Conclusions: ADC-1013 activates splenic dendritic cells as shown by an increase of the co-stimulatory molecules CD80 and CD86. ADC-1013 improves T cell activation as shown by an increase of ICOS and CD44 hi CD62L - effector memory cells. In an OVA vaccination model, ADC-1013 expands OVA-specific T cells and delays growth of an OVA-expressing tumor, demonstrating potential for combination with tumor vaccines. ADC-1013 A single systemic dose of ADC-1013 induces activation of splenic dendritic cells Figure 2 – Effect of ADC-1013 on splenic dendritic cell activation. Mice (hCD40tg) were administered one dose of 100 μg ADC-1013 i.p. and spleens collected for FACS analysis at the indicated time points following treatment. Graph shows percent CD80 + CD86 + cells ± SEM within the CD11c + MHCII + population. Representative FACS plots are shown for the 24 h time point. CD80 CD86 Ctr IgG ADC-1013 ADC-1013 treatment expands antigen-specific T cells in an OVA vaccination model Figure 4 – Effect of ADC-1013 on expansion of OVA-specific T cells. Mice (hCD40tg) were administered 200 μg OVA i.v. and 100 μg ADC-1013 i.p. as shown in Figure 3. Spleens were collected for FACS analysis on day 14. Graph shows percent OVA peptide (SIINFEKL)-MHCI tetramer + cells ± SEM within the CD8 + population. Representative FACS plots from two groups are shown. OVA-MHCI tetramer CD8 Ctr IgG/OVA ADC-1013 every 7 days/OVA ADC-1013 treatment results in improved T cell activation 0 10 20 ADC-1013 every 7 days/OVA ADC-1013 every 2-3 days/OVA Ctr IgG/OVA CD44 hi CD62L - % of CD8 + Figure 5 – Effect of ADC-1013 on splenic T cell activation. Mice (hCD40tg) were administered 200 μg OVA i.v. and 100 μg ADC-1013 i.p. as shown in Figure 3. Spleens were collected for FACS analysis on day 14. Graphs show percent ICOS + (top) and CD44 hi CD62L - (bottom) cells ± SEM within the CD8 + population. Representative FACS plots from two groups are shown for CD44 hi CD62L - cells. CD44 CD62L Ctr IgG/OVA ADC-1013 every 7 days/OVA CD40 ADC-1013 Figure 1 – Effect of systemically administered ADC-1013 on MB49 tumor growth. Mice (hCD40tg) were inoculated with 2.5x10 5 MB49 cells s.c. and administered 100 μg ADC-1013 or Ctr IgG i.p. on days 7, 10 and 13 post-inoculation. Graph shows percent survival from two pooled experiments. World Preclinical Congress 2018 Alligator Bioscience AB, Lund, Sweden
Transcript of Adnan Deronic*, Anneli Nilsson, Mia Thagesson, Doreen ...Adnan Deronic*, Anneli Nilsson, Mia...
Adnan Deronic*, Anneli Nilsson, Mia Thagesson, Doreen Werchau, Niina Veitonmäki, Per Norlén, Christina Furebring and Peter EllmarkAlligator Bioscience AB, Lund, Sweden, *Presenting author
ADC-1013 (JNJ-64457107) is a human CD40 agonistic IgG1 antibody.
It activates antigen-presenting cells (APC) by stimulating CD40 on the surface of e.g. dendritic cells.
Mediated by its effect on APC, ADC-1013 treatment results in activation of tumor-directed cytotoxic
T cells with capacity to eradicate tumors.
Mechanism of action of ADC-1013
ADC-1013 eradicates the syngeneic MB49 bladder tumor in hCD40tg mice
OVA vaccination combined with ADC-1013 treatment delays EG7.OVA tumor growth
Tum
or
volu
me
(mm
3)
Figure 6 – Effect of OVA vaccination combined with prophylactic ADC-1013 treatment on EG7.OVA tumor growth.Mice (hCD40tg) were administered 200 µg OVA i.v. and 100 µg ADC-1013 i.p. as shown in Figure 3 and were inoculated with1x106 EG7.OVA cells s.c. No additional treatments were administered following tumor inoculation. Graphs show tumorvolume ± SEM (left) and percent survival (right) from two pooled experiments.
OVA vaccination model for evaluation of ADC-1013 effect on antigen-specific T cells
Figure 3 – Experimental strategy for the OVA vaccination model. Mice (hCD40tg) were divided in five groups. Threegroups were administered 200 µg OVA i.v. and the remaining two received PBS i.v. on days 0 and 7. Mice were additionallytreated with 100 µg ADC-1013 (white arrows) every 2-3 days or every 7 days as outlined. Ctr IgG (100 µg, grey stippledarrows) was administered to two of the groups as control. On day 14, a cohort of mice from each group was sacrificed forFACS analysis of spleens. The remaining mice were inoculated with 1x106 EG7.OVA cells s.c. and tumor growth evaluated.
ADC-1013 every 2-3 days
Ctr IgG
ADC-1013 every 2-3 days
0 2 5 7 9 12 14
EG7.OVA s.c.
Day:
PBS/OVA i.v.
Ctr IgG
PBS +
OVA +
ADC-1013 every 7 days
Tumor growth
FACS
PBS/OVA i.v.
Summary
IgG1 Fc
CD40-bindingdomain
Aim:To demonstrate the in vivo effect of systemically administeredADC-1013 on APC and antigen-specific T cells.
Methods:Mice transgenic for human CD40 (hCD40tg) were immunizedwith ovalbumin (OVA) and treated systemically with ADC-1013and APC and T cell activation analyzed by flow cytometry. Micewere also inoculated with an OVA-expressing tumor and anti-tumor efficacy of this treatment evaluated.
Conclusions: ADC-1013 activates splenic dendritic cells as shown by an
increase of the co-stimulatory molecules CD80 and CD86. ADC-1013 improves T cell activation as shown by an increase
of ICOS and CD44hi CD62L- effector memory cells. In an OVA vaccination model, ADC-1013 expands OVA-specific
T cells and delays growth of an OVA-expressing tumor,demonstrating potential for combination with tumor vaccines. ADC-1013
A single systemic dose of ADC-1013 induces activation of splenic dendritic cells
Figure 2 – Effect of ADC-1013 on splenic dendritic cell activation. Mice(hCD40tg) were administered one dose of 100 µg ADC-1013 i.p. and spleenscollected for FACS analysis at the indicated time points following treatment.Graph shows percent CD80+ CD86+ cells ± SEM within the CD11c+ MHCII+
population. Representative FACS plots are shown for the 24 h time point.
CD
80
CD86
Ctr
IgG
AD
C-1
01
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ADC-1013 treatment expands antigen-specific T cells in an OVA vaccination model
Figure 4 – Effect of ADC-1013 on expansion of OVA-specific T cells. Mice(hCD40tg) were administered 200 µg OVA i.v. and 100 µg ADC-1013 i.p. asshown in Figure 3. Spleens were collected for FACS analysis on day 14. Graphshows percent OVA peptide (SIINFEKL)-MHCI tetramer+ cells ± SEM within theCD8+ population. Representative FACS plots from two groups are shown.
OV
A-M
HC
Ite
tra
me
r
CD8
Ctr
IgG
/OV
AA
DC
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13
eve
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da
ys/O
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ADC-1013 treatment results in improved T cell activation
0 10 20
ADC-1013 every 7 days/OVA
ADC-1013 every 2-3 days/OVA
Ctr IgG/OVA
CD44hi CD62L-
% of CD8+
Figure 5 – Effect of ADC-1013 on splenic T cell activation. Mice (hCD40tg)were administered 200 µg OVA i.v. and 100 µg ADC-1013 i.p. as shown in Figure3. Spleens were collected for FACS analysis on day 14. Graphs show percentICOS+ (top) and CD44hi CD62L- (bottom) cells ± SEM within the CD8+ population.Representative FACS plots from two groups are shown for CD44hi CD62L- cells.
CD
44
CD62L
Ctr
IgG
/OV
AA
DC
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13
eve
ry7
da
ys/O
VA
CD40
ADC-1013
Figure 1 – Effect of systemically administered ADC-1013 on MB49 tumor growth. Mice (hCD40tg) were inoculated with2.5x105 MB49 cells s.c. and administered 100 µg ADC-1013 or Ctr IgG i.p. on days 7, 10 and 13 post-inoculation. Graphshows percent survival from two pooled experiments.
World Preclinical Congress 2018Alligator Bioscience AB, Lund, Sweden
