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2007/2/8


1.

2.
3.

4.

5.

6.


1. :
a.
b. :
c .
d. :
2. :

3.
4.
3

1999
2001a

4~5,000
4~5,000

2000
2004

1999
2003c
2004d
2005f
2006

a. 200310;
d. UONS/11.20. 2004;
g. UNOS/Jan 14th 2006;

90
120

181
195

3,981
6,774

63,000
83,432
87,268d
91,432
94,757g

8,802
9,433e
13,333
12,394h

17,000
16,984
18,108e
25,952
24,435h

b. Jeong et al., 2006;


c. UONS/11.14. 2003;
e. UONS/11.12. 2004;
f. UONS/Nov. 2005;
h. UONS/Jan Oct, 2006.


1. :

2. :

1. :

2. :

-1, 3-galactosyl transferase


gene evolution mutated

4.

3.

1.

2.

(Nowak R, Science 1994)

(Banz and Rieben, 2006)

hDAF

1.
31
2.
78
3.
(2000)

Animal Cloning Cloned Mini Pigs


May Also Be Organ Donors
Heart and Renal Transplantation
1. Dont undergo HAR
2. Dont require Ab adsorption
3. Dont require complement
inhibition
4. Heart survive 50/60~179 days
5. Kidney survive 30 days
6. Kidney plus thymus survivals
increased to 81 days
1,3-galactosyl transferase KO pig
(November 2002 )
10

(Kuwaki et al., 2005. Nat Med.)

It has being progressing for 12


years on Xenotransplantation,
since November of 1994
In Taiwan
1997Nantes

11

HAL-DP A1+B1
HAL-DQ A1+B1
HAL-DR A1+B1

Y113-7&-8
Y122-02A
D146-13A

85/10/18 /
88/4/21

89/4/30

Y62-11A
Y25-12A
Y25-11A
Y25-13A
Y280-11A
Y15-11A
Y69-10a

91/02/21
92/1/23
91/01/18
91/01/18
91/09/23
92/1/16
93/3/10

Actin-hDAF
hDAF/hHO-1

hHO-1/GFP
12

Resistance
Pressure Sensor

Connecting
Needle
A
U

Isolated
Kidney

Pump

Holding
Funnel

Outlet

Reservoir
O2 and Heat

Filtrated
Collection

Volume Adjustment/
Blood Introduction

The device and facilities of perfusion system


13


Saline perfusion

Human blood perfusion


(40 min)

Saline perfusion

Human blood perfusion

Perfusion Time, (hr:min:sec)

14

Human blood perfusion in insolated kidney of a human DAF


transgenic or non-transgenic pigs. ()

Tg/B(931005)

15

Tg/O (931005)

Tg/A(930929)

SENSTIZATION

Donor
kidney

Passenger leukocyte

Class II MHC
antigen

TH

TH

TH

TH

IL-2
CTL

CTL

CTL

Effector

Lymph node

Migration of passenger leukocyte from a donor graft to regional


lymph nodes of the recipient results in activation of TH cells in
response to different class II MHC antigen expressed by the
passenger leukocytes.
16

HLA-DP Tg Pigs
Responder: HLA DPw4(+) human lymphocyte
*

p=0.08 as compared to SI of NTG

1
0.95

n=4

0.9

SI

0.85

n=4

n=4

0.8
0.75

Human DP4(-)

TG

NTG

Stimulators of MLC

Stimulating indices of direct xenogenic MLC with responder


of HLA DPw4(+) human lymphocytes
17

(JM Lee et al., 2002)

HLA-DQ Tg Pigs
2

1.8
1.6
1.4
1.2

Responder
Human (DQw3-)

0.8
0.6
0.4
0.2
0

18

Human

NP

NTG

TG

Stimulators

Stimulating indices of direct xenogenic MLC with responder of


HLA DQw0601(-) human PBMCs; TG, transgenic pig; NTG,
non-transgenic littermate pig; NP, normal pig. (n=3)
*: p<.05 as compared to TG (SI=1)
#: using the cpm of TG as reference (SI=1)
(JM Lee et al., 2003)

1.6

1.4
1.2

Responder

0.8

Human (DQ+)

0.6
0.4
0.2
0
Human

NP

NTG

TG

Stimulators

Stimulating indices of direct xenogenic MLC with responder of


HLA DQw0601(-) human PBMCs; TG, transgenic pig; NTG,
non-transgenic littermate pig; NP, normal pig. (n=3)
*: p<.05 as compared to TG (SI=1)
#: using the cpm of TG as reference (SI=1)
19

(JM Lee et al., 2003)

*
120
100
80

Responder

60

Human DQw3(-)

40
20
0
Human

NP

TG

Stimulators

INF production in direct xenogenic MLC with responder of HLA


DQw0601(-) human PBMCs after 3 days of stimulation.
TG, ransgenic pig; NP, normal control pig
*: p<.05 as compared to TG (representative one of three studies)
20

(JM Lee et al., 2003)

*
160
140
120
100

Responder

80
60

Human DQw3(+)

40
20
0
Human

NP

TG

Stimulators

TH1 response (INF production) in direct xenogenic MLC with


responder of HLA DQw0601(+) human PBMCs after 3 days of
stimulation.
TG, transgenic pig; NP, normal control pig
*: p<.05 as compared to TG (representative of one three studies)
21

(JM Lee et al., 2003)

Conclusion
This similarity of MHCII between pig and
human by the HLA transgenic pig can help
to reduce the xenogenic cellular response
between human and pig.
Ching-Fu Tu, C.-J. Lee, J. M. Lee, K. Tsuj.
2003. Transgenic swine having HLA-D gene,
swine cells thereof and xenografts there from.
(US 6,639,122 B1/ Patent)
22


Beyond
hyperacute
rejection ?

Hi, I am the first CMV-hDAF transgenic pig


HAR
in Taiwan.

Complement
How many transgenes would you like
Actin-hDAF
to put into my genome?
Natural Ab

Two, four or several?


Delayed-vascular
Protecting Genes
A20
Bcl-2
INB
ATPDase
Heme oxygenase-1
Thrombomodulin

23

1,3-GT KO

SLA

DXR?
HO-1

HAR

HLA-II
Genes
knock out
(Cloning pigs)

Cellular-mediated
SLA-I and II ?
HAL-DR
HLA-DQ
HLA-DP
Fas-L?
HLA-I

Multiple Genetic Modification Model

1()

24

hHO-1/GFP
25


1. Chapman et al., N. Eng. J. Med. 333: 1498-1501. (1995)
2. Patience et al., Nature Med. 3:282-286. (1997)

PK-15
3. Le Tisser et al., Nature 389: 681-682. (1997)
Two sets of human-tropic pig retrovirus. (PERV-A &-B)/PK-15
4. Wilson et al., J. Virol. 72:3082-3087. (1998)
Retrovirus released from porcine primary peripheral blood
mononuclear cells infects human cells.
5. Luc et al., Nature 407:90-94. (2000)

Infection by porcine endogenous retrovirus after


islet xenotransplantation in SCID mice

(Xenotransplantation, 2007)
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