A. B.

Pre
Pre-splenectomy (n=5) Post-splenectomy (n=5)
Post
50
20 20
40

% of total T cells
30
0 0

20

-20 -20 10
tSNE2

tSNE2
0

-20 0 20 -20-20 00 20 20
tSNE1 tSNE1 Colored by phenotype

C. Naïve CD4 T cells

Memory/Effector memory CD4 T cells
Pre
Terminally differentiated CD4 T cells
Naïve CD8 T cells
Post Memory/Effector memory CD8 T cells
Terminally differentiated CD8 T cells
Day: 19 59 110 203 241 Other T cell subsets

D. Pre-splenectomy (n=5) Post-splenectomy (n=5) E. 25

Pre
Post

% of total T cells
20

15

10

0
#21 #22
-20 0 20 Cluster ID

F. #22 Protein Expression

#21 Low High

Tbet

EOMES
CD16

CD11c

CCR7
CD8a
CD33
ICOS
CD11b
CD27
CXCR3
PD1
FoxP3
CD45RO
CCR4
CD123
CCR5
CD7
CD38
CD86
CCR6
CD1c
CD127
PD-L1
CD25
CXCR5
HLA-DR
CD4
CD56

G.
CCR6 CCR4 CD25 CD127 PD1 PD-L1 CD56

4 4.6 2.5 0.8

* 0.6
* 0.5 * 2.0
*
* * * * * * * *
* * *
Relative Expression
Relative Expression

Relative Expression
Relative Expression

Relative Expression

4.4
Relative Expression

*
Relative Expression

2.0 0.4
3 0.6 1.5
4.2 0.4
1.5 * 0.3
2 4.0 0.4 1.0
1.0 0.2
3.8 0.2
1 0.2 0.5
3.6 0.5 * 0.1

0 3.4 0.0 0.0 0.0 0.0 0.0

e

#2 t

#2 t
#2 re

st

#2 re

st
e

#2 t
#2 re

st

#2 t
#2 re

st

#2 re

#2 st
#2 re

st

#2 re

#2 st
#2 re

st

#2 st
#2 re

st
s
s

Pr
Pr

Pr

Pr

Pr
Po

Po
Po

Po

Po

Po

Po

Po

Po

Po

Po

Po

Po

Po
P
P

P
1

2
1

2
#2

2
#2

#2

#2

#2

#2

#2

2
#2
#2

#2

#2

#2