Normal S-CM
6
Cytokine (pg/mL) 500 g total protein / mL S-CM 250 g total protein / mL S-CM 50 g total protein / mL S-CM TGF- 58 + 21 29 + 10.5 6 + 2 IL-6 nd nd nd IL-1 nd nd nd TGF- 76 + 10 38 + 5 7.6 + 1 IL-6 71 + 20 35 + 10 7.1 + 2 IL-1 20 + 8 10 + 4 2 + 0.8 Normal S-CM Crohn’s S-CM nd = not detectible Supplemental Table I. Levels of key immune-modulating cytokines in working concentrations of stroma-conditioned media derived from normal mucosa and inflamed Crohn’s mucosa
description
Supplemental Table I . Levels of key immune-modulating cytokines in working concentrations of stroma-conditioned media derived from normal mucosa and inflamed Crohn’s mucosa. Normal S-CM. Crohn’s S-CM. nd = not detectible. 5. 5. 10. 10. 4. 10. 4. 10. 10. 3. 10. 3. 10. 2. 10. 2. - PowerPoint PPT Presentation
Transcript of Normal S-CM
Cytokine(pg/mL)
500 g total protein /mL
S-CM
250 g total protein /mL
S-CM
50 g total protein /mL
S-CM
TGF- 58 + 21 29 + 10.5 6 + 2
IL-6 nd nd nd
IL-1 nd nd nd
TGF- 76 + 10 38 + 5 7.6 + 1
IL-6 71 + 20 35 + 10 7.1 + 2
IL-1 20 + 8 10 + 4 2 + 0.8
Normal S-CM
Crohn’s S-CM
nd = not detectible
Supplemental Table I. Levels of key immune-modulating cytokines in working concentrations of stroma-conditioned media derived from normal mucosa and inflamed Crohn’s mucosa
+ Normal S-CM
50 g/mL 250 g/mL 500 g/mL
0 102 103 104 105
0102
103
104
105
0 102 103 104 105
0102
103
104
105
40.5
0 102 103 104 105
0102
103
104
105
4.0
0 102 103 104 105
0102
103
104
105
1.4
62.9
0 102 103 104 105
0102
103
104
105
22.9
0 102 103 104 105
0102
103
104
105
35.8
0 102 103 104 105
0102
103
104
105
51.3+ Crohn’s
S-CM
PHA-stimulated
IFN
- (
pg/m
L)
Normal S-CM(g/mL)
Crohn’s S-CM(g/mL)
50 250 5000 50 250 5000
C
B
T-betM
edia
D
-actinN
orm
al
S-C
MC
rohn
’sS
-CM
50 250 500
S-CM (g/mL)
*
**
% In
hibi
tion
of P
rolif
erati
on Normal S-CMCrohn’s S-CM
A
Supplemental Figure 1
Days 0-3: IL-2 + CD3/CD28 or PHA Days 3-7: CFSE + Crohn’s or Normal S-CMDay 7: Harvest cells and measure proliferation and cytokines in supernatant
61.6
17.9
39.1
IL-2 + PHA
64
39
59.8
IL-2 + CD3
19.7
3.3
7.3
IL-2
Media(D. 3-7)
Normal S-CM (D. 3-7)
Crohn’s S-CM (D. 3-7)
Normal S-CM
Crohn’s S-CM
Proliferation
Normal S-CM
Crohn’s S-CMIFN-
p =0.25
p =0.0005
p =0.006
IL-2 CD3 PHA
% In
hib
itio
n o
f IF
N-
Rel
ease
IL-2 CD3 PHA
% In
hib
itio
n o
f P
rolif
erat
ion p =0.021
p =0.052
p =0.035
B
C
A
Supplemental Figure 2
CD3 +
Media(D. 3-7)
Days 0-3: IL-2 + CD3/CD28 or PHA Days 3-7: CFSE + Crohn’s or Normal S-CM + CD3Day 7: Harvest cells and measure proliferation and cytokines in supernatants
IL-2 + PHA66.9
25.5
37.6
IL-2 + CD365.5
44.7
53.9
IL-244.9
5.6
14.3
CD3+
Normal S-CM (D. 3-7)
CD3+
Crohn’s S-CM (D. 3-7)
IL-2 CD3 PHA
% In
hib
itio
n o
f P
rolif
erat
ion p =0.009
p =0.011
p =0.014
Normal S-CM
Crohn’s S-CM
Proliferation
Normal S-CM
Crohn’s S-CMIFN-
p =0.002p =0.007
p =0.0009
IL-2 CD3 PHA%
Inh
ibit
ion
of
IFN
- R
elea
se
C
AB
Supplemental Figure 3
Normal S-CM
Normal S-CM+ -TGF-
(200 g/mL)
0 102 103 104 105
0
102
103
104
105
61.7
0 102 103 104 10 5
0
102
103
104
105
19.9
0 10 2 103 10 4 105
0
10 2
10 3
10 4
10 5
33.3
rhTGF-(pg/mL)
IFN
- (p
g/m
L)
media 5 50 500
rhTGF-(pg/mL)-TGF-(g/mL)S-CM (250 g/mL) + + + +-
- - 2 20 200
6000
3000
2000
1000IFN
- (p
g/m
L)
rhT
GF
- (
pg
/mL
)
5
50
500
C
0 102
103
104
105
<FL1-A>
0
102
103
104
105
<FL16-A>
50.4
0 102
103
104
105
<FL1-A>
0
102
10 3
10 4
10 5
<FL16-A>
56.50 102 103 104 105
<FL1-A>
0
102
103
104
105
<FL16-A>
56.4
0 10 2 103 10 4 105
<FL1-A>
0
10 2
10 3
10 4
105
<FL16-A>
57.6A
BD
Supplemental Figure 4
71.870.4
% C
ells
Pro
lifer
atin
g
-TGF- (100 g/mL)
IgG(100 g/mL)
61 64.1 65.9
-TGF- (100 g/mL)
Purified IgG(100 g/mL)Media
-TGF- (100 g/mL)
IgG(100 g/mL)
Media
IFN
- (
pg
/mL
)
61
66.8
-IL-6(1 g/mL)
-IL-1(1 g/mL)
-IL-6 + -IL-1(1 g/mL)
Media
-IL-6
(1
g/m
L)
-IL-1
(1
g/m
L)-IL
-6
+ -IL
-1(1
g/
mL)Media
% C
ells
Pro
lifer
atin
g
p = 0.06
-IL-6
(1
g/m
L)-IL
-1(1
g/
mL)
-IL-6
+
-IL-1
(1
g/m
L)Med
ia
IFN
- (
pg
/mL
)
A B C
D
E F
Supplemental Figure 5
![PFA(S)[S] and Locally Compact Normal Spaces](https://static.fdocuments.net/doc/165x107/613d4d10736caf36b75bb40e/pfass-and-locally-compact-normal-spaces.jpg)
