CYTOPROTECTIVE EFFECTS OF VIRAL IL10 GENE

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CYTOPROTECTIVE EFFECTS OF VIRAL IL-10 GENE
TRANSFER IN LIVER ISCHEMIA AND REPERFUSION
INJURY ROLE OF INNATE AND ADAPTIVE IMMUNITY
B. Ke, X. Shen, S. Tsuchihashi, F. Gao, J. A.
Araujo, T. Ritter, R. W. Busuttil, and J. W.
Kupiec-Weglinski
The Dumont-UCLA Transplant Center, and
Division of Cardiology, David Geffen School of
Medicine at UCLA, Los Angeles, CA Institute of
Medical Immunology, Berlin, Germany
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INTRODUCTION

ISCHEMIA/REPERFUSION INJURY
Multifactorial process that affects graft
function after liver transplantation. Causes up
to 10 of early organ failure, and can lead to
the higher incidence of both acute and chronic
rejection. Contributes to the acute shortage
of livers.
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BACKGROUND

• TLR4 plays a key role in triggering inflammatory
  response associated with Ag-independent hepatic
  ischemia/reperfusion (I/R) injury. J. Immunol.
  2004 173 7115
• Depression of TLR4 activation or local induction
  of HO-1 is cytoprotective in hepatic I/R injury.
• Transplantation 2003 75 1118, Am. J.
  Transplant. 2003 3 1076, Hum. Gen. Ther. 2004
  15 691
• Interleukin-10 induces expression of heme
  oxygenase-1 (HO-1) with potential
  anti-inflammatory effect. Nat. Med. 2002 8 240

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AIM
To analyze putative cross-talk between innate and
adaptive immune mechanisms in liver I/R injury
following viral IL-10 gene transfer.
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MOUSE PARTIAL LIVER WARM ISCHEMIA MODEL
i. v.
Groups 1) Sham 2) WT (wild-type) 3) WT
Ad-?-gal 4) WT Ad-vIL-10 5) HO-1-/ 6)
HO-1-/ Ad-?-gal 7) HO-1-/ Ad-vIL-10
Ad-vIL-10 or Ad-?-gal (2.5x109 pfu)
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IL-10 ELISA
300
200
pg/ml
100
0
Sham
WT
WT Ad-b-gal
WT Ad-vIL-10
HO-1-/
HO-1-/ Ad- b-gal
HO-1-/ Ad- vIL-10
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sGOT LEVELS (IN VIVO WARM I/R, 6h)
plt 0.005



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HISTOPATHOLOGY (HE Staining)
HISTOPATHOLOGY (HE Staining)
WT Ad-vIL-10
WT
WT Ad-b-gal
1.2 0.5
3.1 0.8
3.40.6
HO-1-/ Ad-b-gal
HO-1-/
HO-1-/ Ad-vIL-10
3.3 0.8
3.5 0.8
1.3 0.5
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MPO ACTIVITY (IN VIVO WARM I/R, 6h)
plt0.05

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5
4
U/gm
3


2
1
0
Sham
WT
WT Ad-b-gal
WT Ad-vIL-10
HO-1-/
HO-1-/ Ad- b-gal
HO-1-/ Ad- vIL-10
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CYTOKINE EXPRESSION
TNF-a
IL-1b
plt0.05

plt0.05





HO-1-/ Ad- b-gal
IL-2
IFN-g
plt0.01

plt0.01





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CHEMOKINE EXPRESSION
E-selectin
MIP-2




plt0.05

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WESTERN BLOTS
HO-1-/ Ad-vIL-10
HO-1-/ Ad-b-gal
WT Ad-vIL-10
WT Ad-b-gal
HO-1-/
Sham
WT
HO-1
A
32 KD
B
Bcl-2
32 KD
C
Bcl-xl
26 KD
D
TLR4
90 KD
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CONCLUSION

• vIL-10 gene transfer prevents hepatic I/R injury
  by downregulating innate TLR4 and modulating the
  adaptive Th1 cytokine and chemokine programs.
• The induction of anti-oxidant HO-1 and
  anti-apoptotic Bcl-2/Bcl-xl by vIL-10 gene
  transfer exert synergistic cytoprotective effect
  against antigen-independent hepatic inflammatory
  injury induced by I/R.