gut and liver cross talk - university of oxford · gut and liver cross talk professor david adams ....
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Oxford Inflammatory Bowel Disease & Hepatology MasterClass
Gut and Liver Cross Talk
Professor David Adams
Gut
Liver
Transport via
portal circulation
• Absorption of food
•Constant exposure to microbes
• Antigen presentation by
professional APC
Strong immune responses
• Metabolic processing of food
•Xenobiotic processing by hepatocytes
• Relatively less pathogen exposure
• Antigen presentation by non
professional APC – endothelium &
hepatocytes
Tolerance
Hepato-enteric immune system
Gut
Liver
Transport via
portal circulation
Hepato-enteric immune system
Co-ordinated immune response to shared
pathogens – a role for mucosal lymphocytes
Trichinella spiralis new born larvae (NBL) and
the hepato-enteric immune response
Liver tolerance to
new borne larvae
Effector CD4 cells
NBL
Trichinella spiralis new born larvae (NBL) and
the hepato-enteric immune response
Liver tolerance to
new borne larvae
Effector CD4 cells Effector CD4 cells
IL-10 deficient
Severe Hepatitis to
new borne larvae
NBL NBL
Trichinella spiralis new born larvae (NBL) and
the hepato-enteric immune response
Liver tolerance to
new borne larvae
Effector CD4 cells Effector CD4 cells No effector CD4 cells
IL-10 deficient
Severe Hepatitis to
new borne larvae
NBL
NBL NBL
Liver tolerance to
new borne larvae
Bliss,SK JI 2007
IL-10 deficient
Inflammatory bowel disease & liver disease
• Primary sclerosing cholangitis or autoimmune hepatitis develop in
– 3-10% of patients with IBD
– 70% of patients with PSC suffer from IBD – Broome et al Dis Col Rec 1995; Loftus Gut 2005
IBD PSC
Diseases
associated with
active bowel
disease
Pyoderma gangrenosum
Pyoderma gangrenosum
Erythema nodosum
Episcleritis
Liver disease runs a course that is independent
of bowel disease
•PSC can develop after colectomy for UC
•UC can develop after liver transplant for PSC
Alabraba et al Liver Tp 2008
Colectomy in the pre or peri-transplant period
protects against recurrence of PSC after liver
transplantation
Colectomy pre liver Tp
Colectomy post liver Tp
No Colectomy
IBD PSC
Do mucosal lymphocytes cause liver disease ?
Hypothesis:
Gut activated lymphocytes in IBD enter the liver and contribute to
liver inflammation in PSC.
Small bowel
Peyer’s patch
DC
migration
CCL25 Madcam-1
Mesenteric LN
Mucosal venule
Adaptive immune responses in the gut
Adams & Eksteen,
Nature Rev Immunol
2006
Small bowel
Peyer’s patch
DC
migration
CCL25 Madcam-1
Mesenteric LN
Mucosal venule
Adaptive immune responses in the gut
Adams & Eksteen,
Nature Rev Immunol
2006
Small bowel
Peyer’s patch
DC
migration
CCR9+ a4b7+
CCL25 MAdCAM-1
Mesenteric LN
Mucosal venule
Adaptive immune responses in the gut
Adams & Eksteen,
Nature Rev Immunol
2006
Imprinting of
a4b7 and
CCR9 confers
gut homing
Small bowel
Peyer’s patch
DC
migration
CCR9+ a4b7+
Mesenteric LN
Mucosal venule
Adaptive immune responses in the gut
Adams & Eksteen,
Nature Rev Immunol
2006
CCL25 MAdCAM-1
CCL25
Small bowel
Peyer’s patch
DC
migration
CCR9+ a4b7+
CCL25 Madcam-1
Mesenteric LN
Mucosal venule
Adaptive immune responses in the gut
Adams & Eksteen,
Nature Rev Immunol
2006
Small bowel
Peyer’s patch
DC
migration
CCR9+ a4b7+
CCL25 Madcam-1
Mesenteric LN
Mucosal venule
Adaptive immune responses in the gut
Adams & Eksteen,
Nature Rev Immunol
2006
CCL25
CCL25 MAdCAM-1
Gut dendritic cells induce expression of a4b7 on T cells
Stagg et al Eur J Immunol 2003
Gut dendritic cells imprint lymphocytes with the
unique mucosal homing phenotype
Selective generation of gut tropic T cells in gut-associated
lymphoid tissue
Johansson-Lindbom,B J Exp. Med 2003
Peyer’s Patch DCs imprint gut homing on lymphocytes
whereas DCs from spleen or PLN cannot
Mora et al Nature 2003
Retinoic acid imprints gut-homing specificity on T cells
Iwata M, Hirakiyama A, Eshima Y, Kagechika H, Kato C, Song SY.
Immunity Oct 2004
•Retinoic acid, enhances expression of a4b7 and CCR9 on T cells
and imprints them with gut tropism.
•Vitamin A deficiency causes a reduction in a4b7 memory T cells
in lymphoid organs and intestinal lamina propria.
Novel role for retinoic acid in imprinting
gut-homing specificity on T cells
Retinoic acid imprints gut-homing specificity on T cells Iwata M et al Immunity Oct 2004
Retinal Retinol Retinoic acid
ADH Retinal
dehydrogenase
Retinoic acid imprints gut-homing specificity on T cells Iwata M et al Immunity Oct 2004
Retinal Retinol Retinoic acid
ADH Retinal
dehydrogenase
Selectively expressed
in gut DCs
Retinoic acid imprints gut-homing specificity on T cells Iwata M et al Immunity Oct 2004
Retinal Retinol Retinoic acid
ADH Retinal
dehydrogenase
RAR
Retinoic acid
receptors
Selectively expressed
in gut DCs
Retinoic acid imprints gut-homing specificity on T cells Iwata M et al Immunity Oct 2004
Retinal Retinol Retinoic acid
RA-responsive
enhancer sequence
CCR9
ADH Retinal
dehydrogenase
RAR
Retinoic acid
receptors
3p21.3
b7 Chromosome 15
Selectively expressed
in gut DCs
MLN DC 71%
Unstimulated 8.8%
Isotype
matched
control mab
CC
R9
Activation of CD3+ PBL at 7 days
CD3/CD28
100nM ATRA
66.4%
(52.6)
Generation of Gut-Homing IgA-Secreting B Cells by
Intestinal Dendritic Cells
Mora, Eksteen et al Science 2006
DCs from human mesenteric LN imprint
CCR9 on responding lymphocytes
Constitutively expressed in the liver
Supports lymphocyte recruitment McNab Gastro 1996; Lalor J Immunol 2002; Bonder et
al Immunity 2005
Increased in mesenteric vessels in IBD Grant 2002; Salmi 1998
Vascular adhesion protein 1 promotes
lymphocyte recruitment to the liver
Contact Arrest
Extravasation
Sinusoidal endothelium
VELOCITY VAP-1
ICAM-1 VAP-1
VAP-1 in normal human liver
VCAM-1 MAdCAM-1
α4β1
α4β7
CCR9
ICAM-1
αLβ2
Gut Endothelial cell
Gut homing lymphocytes Liver infiltrating lymphocytes
VAP-1R
VAP-1
VAP-1R
CXCR3
CXCL10 CCL25
Hepatic Endothelial cell
VAP-1
MAdCAM-1 (VAP-1)
α4β7
CCR9
MLN
MADCAM-1
PORTAL LN
a4b7+ T cells in the
PSC liver
Grant,A.J. et al. 2001 Hepatology.
CD3
α4β7
MAdCAM-1 in PSC
MAdCAM-1
MAdCAM-1
CCL25
α4β7
CCR9
MLN MADCAM-1 and CCL25 upregulated on
endothelium in PSC and autoimmune hepatitis Eksteen et al J Exp Med 2004
CCL25
Merged CCL25
CD31
In PSC 20% of liver lymphocytes express CCR9
+
0 20 40 60 80 100 Mean % CCR9+ T-cells
CD LPL n=3
CD PBL n=3
NL LIL n=6
N PBL n=6
PBC LIL n=6
PSC PBL n=8
PSC LIL n=8
a4b7
CC
R9
PSC LIL 17%
In PSC 20% of liver lymphocytes express CCR9
+
0 20 40 60 80 100 Mean % CCR9+ T-cells
CD LPL n=3
CD PBL n=3
NL LIL n=6
N PBL n=6
PBC LIL n=6
PSC PBL n=8
PSC LIL n=8
a4b7
CC
R9
PSC LIL 17%
In PSC 20% of liver lymphocytes express CCR9
+
0 20 40 60 80 100 Mean % CCR9+ T-cells
CD LPL n=3
CD PBL n=3
NL LIL n=6
N PBL n=6
PBC LIL n=6
PSC PBL n=8
PSC LIL n=8
PBL
IFNγ
PHA 0hrs
PHA 0hrs
PHA 4hrs
PHA 4hrs
1%
13%
78%
2%
These are primed T cells
+CCL25 CCL25 + PTX Control
% c
on
tro
l m
igra
tio
n
80
90
100
110
120
* * p<0.01
*
Miles et al Am J Phys 2008
Transfection of human hepatic sinusoidal endothelial
cells (HSEC) with MAdCAM-1 allows them to support
migration of T cells isolated from PSC liver in the
presence of CCL25 MAdCAM-1
140
What induces the expression of
MADCAM-1 and CCL25 in the
liver?
MAdCAM-1 staining in inflamed PSC liver A. small B. large portal vessels
VAP-1
(SSAO)
RGD motif
Endothelium
Lymphocyte
Amine
Aldehyde
NH3
H2O2
?
Activation
VAP-1:Ligand
interaction
VAP-1 is an adhesion molecule on liver endothelium
It is also an amine oxidase
Amines from
food and wine
and bacteria
Salmi Science 1991; Smith JEM 1998 Jalkanen and Salmi EMBOJ 2001
RGD motif
Endothelium
Lymphocyte
Amine
Aldehyde
NH3
H2O2
?
NFkB Activation
VAP-1:Ligand
interaction
How does enzyme activity affect adhesion?
TRANSCRIPTION OF
CHEMOKINES AND ADHESION
MOLECULES
Lalor et al Hepatology 2007; Liaskou et al Hepatology 2011
MAdCAM-1 is induced on liver endothelium by methylamine & TNFa A
dh
ere
nt
ce
lls
P=0.01 P=0.018
0 10
20 30 40 50 60
Control TNFa TNFa+ TNFa+Meth
Methylamine +anti MAdCAM-1
Liaskou et al Hepatology 2011
Feeding a VAP-1 enzyme substrate methyl amine to mice that over-
express VAP-1 results in increased MADCAM-1 expression in the gut
Liaskou et al Hepatology 2011
Induction of MADCAM-1
Natural amines from gut via portal vein
activate VAP-1
+
Pro-inflammatory cytokines
NFkB activation
MAdCAM-1 induction
VCAM-1 MAdCAM-1
α4β1
α4β7
CCR9
VCAM-1
α4β1
α4β7
MAdCAM-1
Endothelial cell
Gut homing lymphocytes PSC Liver infiltrating lymphocytes
VAP-1R
VAP-1
VAP-1R
CCR9
CCL25 CCL25
Where are gut-homing T cells in PSC
imprinted?
In the liver or the gut?
Neo lymphogenesis is seen in portal-associated
lymphoid follicles in PSC
CCL21 on HEV
CCL19 on
lymphatics and
stromal cells
Grant et al Am J Path 2002
VCAM-1 MAdCAM-1
α4β1 α4β7
CCR9
VCAM-1
α4β1
α4β7
MAdCAM-1
Endothelial cell
Gut homing lymphocytes PSC Liver infiltrating lymphocytes
VAP-1R
VAP-1
VAP-1R
CCR9
CCL25 CCL25
DCs from human liver CANNOT imprint high levels of
functional CCR9 on responding lymphocytes
VCAM-1 MAdCAM-1
α4β1 α4β7
CCR9
VCAM-1
α4β1
α4β7
MAdCAM-1
Endothelial cell
Gut homing lymphocytes PSC Liver infiltrating lymphocytes
VAP-1R
VAP-1
VAP-1R
CCR9
CCL25 CCL25
CCR9+ T cells in the liver are likely to
be mucosal T cells imprinted by gut DC
Under some circumstance T cell activation in the liver
can induce gut homing
Connecting liver and gut: murine liver sinusoidal endothelium
induces gut tropism of CD4+ T cells via retinoic acid.
Neumann K, Kruse N, Szilagyi B, Erben U, Rudolph C, Flach A, Zeitz
M, Hamann A, Klugewitz K. Hepatology. 2012 Jun;55(6):1976-84.
Hepatology. 2012 Jun;55(6):1976-84.
IBD
a4b7
Liver endothelium Gut endothelium
VAP-1, MAdCAM-1 MAdCAM VAP-1
CCL25, CXCR3
Naïve T cell
VAP-1r
MLN + PPs
CCR-9 CXCR3
IBD
a4b7
Naïve T cell
Memory T cell
VAP-1r
MLN + PPs
CCR-9
Liver endothelium Gut endothelium
VAP-1, MAdCAM-1 MAdCAM
CCL25, CCL28
GUT
a4b7
Liver endothelium Gut endothelium
VAP-1, MAdCAM-1 MAdCAM
NO antigen
NO retention
release to lymphatics
RECIRCULATION
Naïve T cell
Memory T cell
VAP-1r
MLN + PPs
a4b7 VAP-1r
Naïve T cell
Memory T cell
MLN + PPs
+antigen
activation
retention
INFLAMMATION
a4b7 VAP-1r
CCR-9
a4b7 VAP-1r
CCR9
GUT
VAP-1, MAdCAM-1 MAdCAM
Naïve T cell
Memory T cell
MLN + PPs
GUT
Liver endothelium Gut endothelium
MAdCAM-1 CCL25 MAdCAM, VAP-1
a4b7 VAP-1r
MAdCAM
CCL25
CCL28
CCR9
+antigen
activation
retention
INFLAMMATION
a4b7 VAP-1r
CCR9 CXCR3
Naïve T cell
Memory T cell
MLN + PPs
GUT
Liver endothelium Gut endothelium
MAdCAM-1 CCL25 MAdCAM, VAP-1
a4b7 VAP-1r
MAdCAM
CCL25
CCL28
CCR9
+antigen
activation
retention
INFLAMMATION
a4b7 VAP-1r
CCR9 CXCR3
Inhibitors of CCR9 and a4b7
currently in clinical trials for IBD
Will they work in PSC?
CREDITS
Collaborators
Sirpa Jalkanen and Marko Salmi (Turku)
Ron Gladue and Nick Pullen (Pfizer)
Mike Briskin (Merrimack Pharma)
Uli von Andrian and Rodrigo Mora (Harvard)
THE UNIVERSITY OF BIRMI NGHAMBirmingham
Liver Labs
Bertus Eksteen
Allister Grant
Trish Lalor
Lia Liaskou
Palak Trivedi
Stuart Curbishley
Alice Miles
Chris Weston
Dept Pathology
Stefan Hubscher