1. Table 1 Case # Age at primary Biopsy (year) Age at metastasis
Primary Tumor
site
Metastasis
Biopsy
decalcification
PD-L1 metastatic
expression
PD-1+ TIL metastatic 1 7 9
R radius
L iliac bone & soft tissue No + 2 8
L distal femur
Mediatinum, thymus
Yes 3 14 18
R distal femur
Lung - 4 12 ++ 5 15 6 13
R femur
L proximal tibia 10
L humerus 21
R shoulder 11
N/A
Unknown 16
Table 1

2. Metastatic Osteosarcoma
Primary Osteosarcoma
Metastatic Osteosarcoma A.
50um
50um
50um
PD-L1 C.
50um
50um
Isotype B.
50um
50um
50um
50um
PD-1
Figure 1A. PDL1 staining
50um
50um
50um
Isotype

3. Days post implantation
%Survival
P<0.0001
Figure 2, CD8 depleted (dotted), control (solid)

4. A. C. B. D. PD-L1 % of Maximum 30% 4073 1848 CD8 PD-1 SPLEEN TIL
Pre Implantation
Tumor C.
CD8
PD-1
SPLEEN
TIL 6%
87%
30%
94%
13% B. D.
Isotype
PD1
Healthy Lung
Late Disease Lung
Early Disease Lung
50 µm *
%PD1+ CD8
Figure 3
50 µm
SPLEEN
TIL

5. A. C. B. D. * * * * * * * * * * * * * * Figure 4A, TIL CD8 p=0.0046
TIL + 4T1
TIL + 4T1
TIL + K7M2
TIL + K7M2 B.
TIL + K7M2
PD-L1 shRNA
TIL + 4T1
K7M2 lysate
TIL + K7M2
PD-L1 shRNA
TIL + 4T1
K7M2 lysate D. * * * * * *
Figure 4A,
TIL CD8 p=0.0046
TIL CD8, tumor lysate p=0.021
TIL + Antigen in tumor versus lysate p=0.0437
TIL + K7M2 compared to TIL +K7m2 PD-L1- p=0.001 B.
TIL CD8 differences p=0.009
TIL CD8+ K7M2 PDL1- differences p=0.05
Tumor lysate differences p=
Tumor lysate compared to TIL k7m2 p=0.02
TIL K7M2 compared to TIL K7M2 PDL1 – p=0.002
C. tIL K7M2 differences p= 0.009
K7M2 PD-L1- differences = .109
Tumor lysate differences =.8718
TIL K7M2 compared to tumor lysate p=0.009
TIL K7M2 compared to K7M2 PD-L1- p=0.0107
Open bars=TIL only
Black bars= TIL+Antigen
D. K7M2 compared to 4T1 with K7M2 p<0.0001
K7M2 compared to K7M2 PD-L1- p<0.0001
TIL
TIL
TIL + 4T1
TIL + K7M2
TIL + K7M2
PD-L1 shRNA
TIL + 4T1
K7M2 lysate

6. A. B. %Survival P=0.0005 Days post implantation
Figure 5A.
Solid line= controls
Dashed= PD-L1 blockade
Volume of largest metastasis (mm3)
# of metastases
Control
α-PD-L1 Ab
Control
α-PD-L1 Ab

7. A. C. B. %IFNγ+ Control PD-L1 Ab %IL-2+ Control PD-L1 Ab %TNF+ Control*
%IL-2+ *
Control
PD-L1 Ab
%TNF+ * B.
Figure 6
Control
PD-L1 Ab