LFA-1/ICAM-1 ligation in T-cells
influences both Notch and
TGF-β pathways
Navin K. Verma
[email protected]
T-cell migration into inflamed tissues
N Engl J Med 1999; 341:1817-28.
T-cell migration in health and diseases
Regulated T-cell
migration
Immune response
regulations
Uncontrolled T-cell
trafficking
Chronic
inflammatory
diseases
Laboratory model of T-cell migration
Resting T-cells
LFA-1
ICAM-1
Migrating T-cells
Receptor
activation
Ong et al, 2014, J Biol Chem; Volkov et al, 2001, Nat Immunol
4
LFA-1/ICAM-1: changes in gene expression
TGF-β pathway
Notch pathway
Verma et al. (2012) J Biol Chem
LFA-1/ICAM-1 up-regulates both TGF-β inhibitory
molecules and Notch signalling proteins
(verified by RT-qPCR)
6
a. SMAD7
5
*
b. SMURF2
5
6
4
3
3
3
2
2
1
1
1
0
0
0
*
0
4
1
3
4
e. NOTCH1
*
3
1
3
f. JAG1
6
*
3
0
4
1
3
3
*
2
2
1
1
1
0
0
0
1
3
6
2
0
1
3
6
*
0
1
3
18
16
14
12
10
8
6
4
2
0
6
g. JAG2
*
0
*
*
2
*
0
6
c. SKI
5
4
4
mRNA levels (fold change)
6
*
6
d. SKIL
*
TGF-β
pathway
*
0
140
120
100
80
60
40
20
0
1
3
6
h. HEY1
*
Notch
pathway
*
0
1
3
6
Duration of T-cell migration on ICAM-1 (h)
Verma et al. (2012) J Biol Chem
Notch Pathway: LFA-1/ICAM-1 activates Notch signalling
+ GSI
Actin
Merge
Cleaved NICD
Nuclei
ICAM-1 (30min)
PLL
Nuclei
pCBF1-Luc reporter
Jag1
Actin
Relative Luciferase Unit
Merge
Cleaved NICD
30
ICAM-1
25
*
Jag1 + DAPT
20
ICAM-1 + DAPT
*
*
*
15
*
*
*
10
5
0
Time of incubation (min)
Relative nuclear flourescence
intensity of cleaved NICD
40
*
30
* *
*
- GSI
+ GSI
*
*
*
3
20
10
1
0
0
60
120
180
240
300
360
2
Duration of T-cell incubation on ICAM-1 (min)
Unpublished data
LFA-1/ICAM-1 activates Notch signalling
Time on ICAM-1 (h)
0
1
3
6
12
GSI
ICAM-1
-
+
+
-
+
+
Control IgG
ICAM-1
NICD
NICD
NICD
Hey1
Hey1
Hey1
Hes1
α-Tubulin
Hes1
α-Tubulin
-
+
α-LFA-1
-
+
Hes1
α-Tubulin
3
Unpublished data
LFA-1 regulates Notch via Akt/ERK/GSK3β
signalling axis
Relative Luciferace Unit
25
+IgG
+ICAM-1
20
*
15
10
5
0
-
Wort
ERKi
LiCl2
LFA-1/ICAM-1 favors Notch-dependent Th1 polarization
10
Unpublished data
Notch Pathway summary
TGF-β pathway: LFA-1/ICAM-1 up-regulates TGF-β
inhibitory molecules
Verma et al. (2012) J Biol Chem
ICAM-1
- + - + -
JNK-I
ST3 pep
AG490
Control
LFA-1/ICAM-1-induced TGF-β inhibitory pathway is
dependent on STAT3 and/or JNK
+ - +
SMAD7
SMURF2
SKI
β-Actin
Verma et al. (2012) J Biol Chem
LFA-1 stimulated T-cells are refractory to iTreg or Th17 differentiation
Mouse
- ICAM-1
b
- TGF-β
*
50
40
30
10
3
2
10
2
1
10
1
10
3
10
10
3.88
+ ICAM-1
2.87
20
10
1
2
3
4
-
+
+
-
+
+
Mouse
50
0
0
*
0
+ TGF-β
0
CD4
10
1
10
2
10
3
0
10
3
2
10
2
1
10
1
10
3
10
10
41
10
1
10
2
10
3
20.4
40
30
20
10
0
0
0
ICAM-1
TGF-β
1
2
3
4
-
+
+
-
+
+
%ROR-g t +/CD4+ T-cells
Th17
d
0
10
Human
50
e
2
10
3
0
10
1
10
2
10
3
Human
*
30
20
10
ICAM-1
TGF-β
10
2500
*
40
0
1
FOXP3 eGFP
IL-17A (pg/ml)
%FOXP3+/CD4+ T-cells
c
Human
60
ICAM-1
TGF-β
%FOXP3+/CD4+ T-cells
iTreg
a
1
2
3
4
-
+
+
-
+
+
2000
1500
1000
500
0
ICAM-1
TGF-β
1
2
3
4
-
+
+
-
+
+
Verma et al. (2012) J Biol Chem
Summary: LFA-1/ICAM-1 induced TGF-β refractoriness in T-cells
LFA-1/ICAM-1 signalling
TGF-β signalling
ICAM-1
TGF-β
LFA-1
TGF-β receptor
P
JNK
P
P
SMAD2
SMAD7
SMAD3
SMURF2
P
P
SMAD2
SMAD3
SKI
P
P
c-Fos
P
c-Jun
P
P
P
SMAD2
SMAD3
AP-1
SMAD7
SKI
SMURF2
Inhibition of IL-2 secretion, Th17, iTreg
Unpublished illustration
Laboratory members
Fazil
Jianhui
Carey
Grace
Navin
Madhavi
Amutha
Seow
Praseetha
Existing collaborations
Funding support
New
collaborations