0

10

20

30

40

50

60

70

80

90

FPKM

of C

D28

rela

tive

to T

-cel

l sig

natu

re

ICO

S-CD

28 fu

sion

D

124V

AITL Normal

D12

4E +

T19

5P

T195

P

TFH Teff Naïve CD4+

p<0.0001

T195

P

A

B C D

A.

Case 089 ACT>CCT p.T195P

Case 086 GAC>GAG p.D124E

B. C.

Case 086 ACT>CCT p.T195P

CD80 IgV-containing Ligand bound A B C

D124

L141

V124

D E F

G H I

-5.00

5.00

CTLA4

CD80 K130

L141 CD28 WT

CD80 K113

D124

K127

CD28 D124V

CD80 K113

V124 K127

J K L

M N O

L141 D124 V124

CTLA4 CD28 W

T CD28 D124V

R94 R94 E81 E81 R94 E81

K20 K20

Supplemental Figure 3

CD28 WT

CD28 mutant

p<0.001p<0.007

p<0.009p<0.04

p<0.03p<0.004

p<0.002p<0.04

A B C

D E

F G

H I

Supplemental Figure 4