(a) (b) supplementary figure s1. fsc-a live/dead cd3 nk1.1 cd4 cd8 4-1bb pd-1 4-1bb pd-1...
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(A)
(B)
Supplementary Figure S1
FSC-A
Live
/Dea
d
CD3
NK1
.1
CD4
CD8
4-1BB
PD-1
4-1BB
PD-1
CD4+ CD8+CD3+Live cells(A) Isotype-treated
FSC-A
Live
/Dea
d
CD3
NK1
.1
CD4
CD8
4-1BB
PD-1
4-1BB
PD-1
CD4+ CD8+CD3+Live cells(B) Anti-4-1BB-treated
Supplementary Figure S2
Supplementary Figure S3
(A) (B) (C)
Supplementary Figure S4
Supplementary Figure S5
Isoty
pe Ctrl
4-1B
BPD-1
4-1B
B/PD-1
0
200
400
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800
1000 Platelets
Pla
tele
ts (
1,00
0/
L)
Isoty
pe Ctrl
4-1B
BPD-1
4-1B
B/PD-1
0
5000
10000
15000 Lymphocytes
Ly
mp
ho
cy
tes
(
L)
Isoty
pe Ctrl
4-1B
BPD-1
4-1B
B/PD-1
0
200
400
600
800 Neutrophils
Neu
tro
ph
ils (
L)
Isoty
pe Ctrl
4-1B
BPD-1
4-1B
B/PD-1
0
100
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AL
T (
U/L
)
ALT
*
Supplementary Tables
Table S1A Incidence and Severity of Microscopic Findings in the Liver
Group 1 (PBS), Group 2-4 (0.1, 1, or 5 mg/kg 4-1BB agonist mAb, respectively), Group 5 (20 mg/kg PD- antagonist mAb), Group 6-8 (20 mg/kg PD-1 antagonist mAb plus 0.1, 1, or 5 mg/kg 4-1BB agonist mAb, respectively) mice were euthanized for necropsy and histological analysis 10 days after a single subcutaneous administration.
Microscopic finding Group 1 Group 2 Group 3 Group 4 Group 5 Group 6 Group 7 Group 8
Mixed Cell Infiltration 3/5 4/5 5/5 5/5 4/5 5/5 5/5 5/5
Minimal 3 4 - - 4 4 - -
Mild - - - 2 - 1 1 1
Moderate - - 5 3 - - 4 3
Marked - - - - - - - 1
Single Cell Necrosis 0/5 0/5 5/5 5/5 0/5 0/5 5/5 5/5
Minimal - - 5 3 - - 4 3
Mild - - - 2 - - 1 2
Supplementary Table S1A
Table S1B Incidence and Severity of Microscopic Findings in the Spleen
See Table 1a for group assignments. * Extramedulary hematopoiesis
Microscopic Findings Group 1 Group 2 Group 3 Group 4 Group 5 Group 6 Group 7 Group 8
Lymphoid Hyperplasia 1/5 3/5 4/5 4/5 5/5 5/5 5/5 5/5
Minimal 1 3 3 3 3 3 1 -
Mild - - 1 1 2 2 4 4
Moderate - - - 1
Increased EMH* 2/5 2/5 2/5 2/5 1/5 0/5 5/5 5/5
Minimal 2 2 2 1 1 - 4 4
Mild - - - 1 - - 1 1
Supplementary Table S1B