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Jun 21, 2017 -
Euthanasia do
d21
.
21
days. Intra-oral. Tissue atraumatic collection. MW -- ...
GJ18d1
. IB13 d2. DF98 d2.
GJ18
da. IB13 d7. DF98 d7.
GJ18
d7 .... 0 o 2 4 6 8 10
20
30 40 50.
19
. 0 2 4 6 8 10
20
30 40 50. MIF. FGF. HGF. 3000.
Buccal
pfu/ml
1000 800
4000
600
2000
GK59 - 106 HB59 - 106 JK74 - 106 GK59 - 105 HB59 - 105 JK74 - 105 GK59 - 104 HB59 - 104 JK74 - 104
1000
400 200
500
0
0
0
5
10
Days
Supplementary Figure 1
Tonsil
6000
15
0
5
10
Days
15
ACV - 106
B
A F
d0
Euthanasia
200 bp
d21
100 bp
Tissue collection
200 bp
21 days Intra-oral atraumatic
MW --
EL75 BL
EL75 d3
EL75 d7
MW --
FB90 BL
FB90 d3
FB90 d7
MW --
EL75 d14
FB90 d14
C MW --
EL75 TG
FB90 TG
EL75 PT
MW --
200 bp
--
Oral swab
Supplementary Figure 2
MW --
Oral swab
EL75 FB90 EL75 FB90
MW --
EL75 d14 EL75 d21 FB90 d14 FB90 d21
300 bp 200 bp
water
D
EL75 BL EL75 d3 EL75 d7 FB90 BL FB90 d3 FB90 d7
100 bp
TG
PT
--
FB90 PT
100 vp 10 vp 100 vp 10 vp
100 bp
MW --
EL75 d21
FB90 d21
GJ18 d2
10 vp MW
IB13 d35
DF98 d35
GJ18 d35
10 vp
MW
IB13 d3
DF98 d3
GJ18 d3
10 vp MW
IB13 d42
DF98 d42
GJ18 d42
10 vp 200 bp
200 bp 100 bp
MW
IB13 d14
DF98 d14
GJ18 d14
10 vp MW
MW
IB13 d21
DF98 d21
GJ18 d21
10 vp
IB13 d49
IB13 d56
IB13 TG
DF98 TG
GJ18 TG
Supplementary Figure 3
10 vp
GJ18 d49
10 vp
DF98 d56
300 bp 200 bp GJ18 d56 10 vp 100 bp
300 bp 200 bp 100 bp
D MW
DF98 d49
MW --
--
MW --
MW --
-- --
-- --
--
IB13 d28 DF98 d28 GJ18 d28
10 vp MW
--
IB13 d42 DF98 d42 GJ18 d42
GJ18 d7
water
DF98 d7
C 200 bp 100 bp
water
water
IB13 d7
10 vp
200 bp 100 bp
MW
IB13 TG DF98 TG GJ18 TG
200 bp 100 bp
MW -- 10 vp
100 bp
water
200 bp 100 bp
300 bp
MW --
MW --
MW --
-- --
--
-- --
--
--
MW --
-- --
--
IB13 d56 DF98 d56 GJ18 d56
DF98 d2
water
IB13 d2
MW water
10 vp
MW
MW water
10 vp
200 bp 100 bp
300 bp 200 bp
IB13 d3 DF98 d3 GJ18 d3
10 vp
IB13 d21 DF98 d21 GJ18 d21
GJ18 d28
IB13 d35 DF98 d35 GJ18 d35
DF98 d28
water
IB13 d28
water
10 vp MW
10 vp
GJ18 d1
10 vp
DF98 d1
IB13 d49 DF98 d49 GJ18 d49
IB13 d1
water
MW
200 bp 100 bp
IB13 d14 DF98 d14 GJ18 d14
B
IB13 d1 DF98 d1 GJ18 d1 IB13 d2 DF98 d2 d2 GJ18 d3 IB13 d7 DF98 d7 GJ18 d7
A
MW
B
40
Run 1 Run 2 Run 3
35
100 genomes
10 genomes
1 genome
CT
1000 genomes
A
30
Efficiency % = 101.7 ± 4.7
25 20 100 101 102 103 104 105
Genomes
C HSV-1 gD IB13 24h swab IB13 TG
Supplementary Figure 4
40
IL-1β
1500
150
20
100
10
1500
IL-2
IL-12
IFN-γ
60
200
30
0
250
1000
40
500
20
50 0 2 4 6 8 10
20 30 40 50
CCL2
0
60
0 2 4 6 8 10
20 30 40 50
CCL3
55
1000
0
60
0 2 4 6 8 10
20 30 40 50
CCL4
0
0 2 4 6 8 10
CCL5
15000
40
10000
20
5000
20 30 40 50
50
500
pg/ml
0
800
45
0 2 4 6 8 10
20 30 40 50
CCL11
40
1500
0 2 4 6 8 10
20 30 40 50
CCL22
1000
CXCL8
20 30 40 50
MIF
0
40
0 2 4 6 8 10
20 30 40 50
FGF
0 2 4 6 8 10
20 30 40 50
20 30 40 50
20 30 40 50
0
0 2 4 6 8 10
20 30 40 50
days post HSV-1 infection
Supplementary Figure 5
19
0 2 4 6 8 10
IB13 DF98 GJ18
200 0 2 4 6 8 10
GM-CSF
23
HGF
400
10 0
0 2 4 6 8 10
600
20
1000
1000
20 30 40 50
20
800
30
2000
0
0 2 4 6 8 10
21
5 0 2 4 6 8 10
0
22
10
500
200
0
20 30 40 50
15
400
3000
25
0 2 4 6 8 10
20
600
0
0
20 30 40 50
all HSV-1 T cell flow_170407.jo
10
3
10
2
1.64
10
4
10
3
10
2
0
4
10
3
10
2
2
3
4
10 10 : CD95
5
10
10
5
10
4
10
3
10
2
31.7
9.53
0
3
5
10
4
10
3
10
2
10
3
10
2
2
3
4
10 10 : CD95
5
10
5.12
10
5
10
4
10
3
10
2
4
10
3
25.5
10
2
0 10
2
3
CD95
2
4
5
10
3
4
10 10 : CD95
5
10
3
4
10
4
10
4
10
10 10 : CD95
5
0.231
0
0 10
10
30.4
0
10 10 : CD95
2
0
5
10
0
0 10
19.7
0 10
10
0
Supplementary Figure 6
4
10 10 : CD95
5
10
: CCR6
2
: CCR6
10
4
0
0 10
CCR6
10
0
0 10
: a4b7
10
: a4b7
α4β 7
5
5
0
0 10
10
2.19
10
: a4b7
4
5
Control
0 10
2
3
2
4
10
3
10
2
4
5
10
5
0.38
0
10 10 : CD95
3
10 10 : CD95
5
10
: CCR6
10
10
: CCR5
5
: CCR5
CCR5
10
Day 2 : CCR5
Day 0
new supp fig
0 10
2
3
10 10 : CD95
5
20170122_pilot 2 HSV ICS_170614
3
4
10 5
3
10
2
10
2
3
4
0 10 10 10 : CD69
10 5
0
10
4
10
3
10
2
0
10
3
4
0 10 10 10 : CD69
C 250K
200K
2
5
2
3
10 5
10
4
10
3
10
2
200K
%
0.3
91.8
10
5
2
3
4
0 10 10 10 : CD69
0.0
81.3
0.4
150K
0.3 0.2
100K
UL39
US6 HSV-1
%
0 10
0.0
P/I
3
4
10 10 : live/dead
%
10
3
10
2
10
4.84
100K
60 10 40 20
5
0.5 10
4
US6 HSV-1
0.3
0.2
0.2
0.0
P/I
10 2 0 UL19
UL39
US6 HSV-1
8.24 P/I
10
2
0
5
2
3
4
0 10 10 10 : CD69
10 5
10
4
10
3
10
2
10
5
1.03
10
2
3
4
0 10 10 10 : CD69
10 5
10
4
10
3
10
2
10
5
3.58
0
5
2
3
4
0 10 10 10 : CD69
10
5
2
3
4
0 10 10 10 : CD69
10
5
IB13 DF98 GJ18
0.1
10
560
2
0 10
40 20
3
10 10 : CD3
4
10
5
60 40 20
2
3
4
0 10 10 10 : CD4
5
10
0.0
UL19
UL39
US6 HSV-1
P/I
60 40 20 0.5 0.4
0.3
0.3
0.2
0.2
0.2
0.2
0.1
0.1
0.1
0.0
0.0
0.0
UL39
US6 HSV-1
P/I
UL19
UL39
US6 HSV-1
P/I
0.4
0.5 0.4 0.3 0.2 0.1 P/I
0.0
0.5
0.5
0.4
0.4
0.3
0.3
0.3
0.2
0.2
Page 1 of 1
US6 HSV-1
P/I
0.0
UL19
UL39
US6 HSV-1
0.0
30 20 10
60 40 20
20 15 10 5
0.5
0.5
0.4
0.4
0.4
0.3
0.3
0.3
0.2
0.2
0.2
0.1
0.1 UL39
US6 HSV-1
P/I
0.0
P/I
IB13 DF98 GJ18
(FlowJo v9.9.5)
0.1 P/I
0.5
UL19
US6 HSV-1
0.2
0.1 UL39
UL39
IL-17
0.4
UL19
UL19
TNF-α
0.5
0.0 US6 HSV-1
P/I
20 15 10 5
P/I
60 40 20
US6 HSV-1
60 40 20
0.0 US6 HSV-1
UL39
80 60 40 20
0.1
0.1
IB13 DF98 GJ18
0.1 UL19
IL-2
30 20 10
UL39
3
: IL2
: IL2
0.4
0.3 10 3
0.4
UL19
2
10
4.75
0.5
0.4
0.1 50K
67.2
0.5
0.0
10
10
4
IL-17
0.3
UL39
10
3
10 5
60 40 20
0.4
UL19
10
TNF-α
150K
UL39
4
0 10 10 10 : CD69
200K
UL19
3
0.3
0.0
10
4
1.19
0
0.4
0.2
%
4
0.5
0.3
Supplementary Figure 7
10
0.5
UL19
10 5
0 2
60250K 40 20
0.5
SPLN
4
0.425
2
0.1 50K
UL19
3
10 5
5
0.5
IFN-γ
6/21/17 10:35 AM
2
CD8
PMA/iono
0
0 10 10 10 : CD69
IL-2
0 200K
PBMC
2
5
250K
60 40 20
0.1
50K
SPLN
10
0
250K
60 40 20
0.2
100K 150K FSC-A
10
2.38
IFN-γ
PBMC
100K
50K
4
0 10 10 10 : CD69
0.4
0
4
0
150K
0
3
0 10 10 10 : CD69
0
0.5 FSC-H
gating strategy PBMC
10
10
3
0.726
0
5
0
2
10
4
0
5
: IL2
10
: IL2
4
2
FSC-H
2
10
10
0
0 10 10 10 : CD69
CD8
10
3
UL39
10 5
: IL2
2
10
4
DMSO
CD4
5.36
: CD8
10
0
10 5
: IL2
2
10
3
0.427
FSC-H
10
10
4
: IL2
10
3
: IL2
10
4
10 5
0
: IL2
10 5
B
PMA/iono
: IL2
UL39
: IL2
DMSO
SPLN
: IL2
PBMC
A CD4
Layout: IL2 secreted by GJ18 PBMC-1
UL19
UL39
US6 HSV-1
P/I
IB13 DF98 GJ18
0.1 UL19
UL39
US6 HSV-1
P/I
0.0
UL19
UL39
US6 HSV-1
P/I
20170122_pilot 2 HSV ICS_170614
2
3
4
0 10 10 10 : CD4
4
10
3
10
2
2
0
3
4
4
10
3
10
2
5
3
4
0 10 10 10 : CD4
5
10
Supplementary Figure 8
3
4
0 10 10 10 : CD4
10
4
10
3
10
2
5
3
4
0 10 10 10 : CD4
5
10
3
4
0 10 10 10 : CD4
10
2
10
4
10
3
10
2
5
3
4
0 10 10 10 : CD4
5
10
3
4
0 10 10 10 : CD4
10
3
10
2
10
4
10
3
10
2
5
3
4
0 10 10 10 : CD4
5
10
3
4
0 10 10 10 : CD4
4
10
3
10
2
10
4
10
3
10
2
5
3
4
0 10 10 10 : CD4
5
10
3
4
10
4
10
0 10 10 10 : CD4
10
4
10
3
10
2
5
45.8
10 5
0
2
2
10
0.441
10 5
0
2
10
0
2
10
0.346
10 5
0
2
4
55.2
10 5
0
2
10
0.524
10 5
0
2
3
10
0
2
10
0.713
10 5
0
2
2
10
0
2
10
0.554
10 5
10
10
4
0.391
10 5
: TNFa
2
3
0
0 10 10 10 : CD4
: TNFa
10
5
10
0.603
10 5
: TNFa
CD8
10
0
10
10
PMA/iono
: TNFa
2
3
4
0.26
10 5
: TNFa
10
0
10
10
UV HSV-1
: TNFa
2
3
4
0.682
10 5
: TNFa
10
10
10
UV mock
: TNFa
3
4
: TNFa
10
10
US6
0.74
10 5
: TNFa
4
0.818
10 5
: TNFa
10
UL39
: TNFa
0.68
10 5
: TNFa
CD4
UL19
: TNFa
DMSO
Layout: Paper Figure: TNF secreted by DF98 P
0
2
3
4
0 10 10 10 : CD4
5
10
2
3
0 10 10 10 : CD4
5
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