© 2000 American Heart Association, Inc.
Vascular Biology |
Mouse Model of Transplant Arteriosclerosis
Role of Intercellular Adhesion Molecule-1
From the Institute for General and Experimental Pathology (H.D., Y.H., G.W.), University of Innsbruck Medical School; the Institute for Biomedical Aging Research (Y.Z., G.W., Q.X.), Austrian Academy of Sciences; and the Departments of Pathology (S.D.) and Internal Medicine (R.K.), University of Innsbruck Medical School, Innsbruck, Austria.
Correspondence to Dr Qingbo Xu, Institute for Biomedical Aging Research, Austrian Academy of Sciences, Rennweg 10, A-6020 Innsbruck, Austria. E-mail Qingbo.Xu{at}oeaw.ac.at
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Abstract |
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Abstract—Transplant-accelerated arteriosclerosis in coronary arteries is the major limitation to long-term survival of patients with heart transplantation. The pathogenesis of this disease is not fully understood. Herein, we describe a simplified model of artery allografts in the mouse that allows us to take advantage of transgenic, knockout, or mutant animals. Common carotid arteries or aortic vessels were end-to-end allografted into carotid arteries between C57BL/6J and BALB/c mice. Neointimal lesions were observed as early as 2 weeks after surgery and had progressed at 4 and 6 weeks postoperatively. The lumen of grafted arteries was significantly narrowed due to neointima hyperplasia 4 weeks after transplantation. Using this model, we studied the role of intercellular adhesion molecule-1 (ICAM-1) in the development of transplant arteriosclerosis in ICAM-1–deficient mice. Neointimal lesions of artery grafts from ICAM-1 –/– C57BL/6J to BALB/c mice were reduced up to 60% compared with wild-type controls. MAC-1 (CD11b/18)-positive cells adhering to the surface of ICAM-1 –/– artery grafts were significantly less as identified by en face immunofluorescence, and these positive cells were more abundant in intimal lesions of artery grafts in wild-type mice. Furthermore, the major cell component of neointimal lesions 4 weeks after surgery was found to be
-actin–positive
smooth muscle cells, which were significantly reduced in lesions of
ICAM-1 –/– artery grafts. Thus, this model has been proven to be
useful for understanding the mechanism of transplant
arteriosclerosis. Our findings demonstrate that
ICAM-1 is critical in the development of allograft
arteriosclerosis via mediation of leukocyte
adhesion to, and infiltration into, the vessel wall.
Key Words: transplant arteriosclerosis • mouse models • ICAM-1 deficiency
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Introduction |
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Allograft accelerated-transplant arteriosclerosis is the main limitation to long-term survival of patients with organ transplantation.1 2 A hallmark of lesions is mononuclear cell infiltration into the vessel wall of grafts at the early stage, followed by neointimal formation, which largely constitutes smooth muscle cells (SMCs).3 Although the pathogenesis of the disease remains to be studied, it has been observed that the earliest cellular event is leukocyte adhesion to the endothelial surface of the vessel wall within transplanted organs. Subsequently, T cells and monocytes infiltrate into the arterial wall.4 However, the molecular mechanisms by which leukocytes are continuously recruited to the vessel wall of grafted organs in vivo is not fully elucidated.
Intercellular adhesion molecule-1 (ICAM-1), a surface
glycoprotein of the immunoglobulin superfamily, contains 5
immunoglobulin-like motifs in its extracellular domain, followed by a
single transmembrane region and a short cytoplasmic
tail.5 6 Several lines of evidence suggest that
ICAM-1/MAC-1–dependent cellular interaction is involved in a number of
inflammatory processes and in arteriosclerosis via
mononuclear cell adhesion and migration.7 8 9 10 ICAM-1 is a
counterreceptor for the ß2 leukocyte integrins MAC-1
(Mß2, CD11b/CD18) and LFA-1
(Lß2, CD11a/CD18), and their engagement
results in leukocyte adhesion and transmigration through the
endothelium.11
Attracted by the well-defined genetic systems, a number of investigators have begun to use the mouse as an experimental system for transplant arteriosclerosis research.12 13 14 15 Hundreds of inbred lines have been established, the genetic map is relatively well-defined, and both congenic strains and recombinant strains are available to facilitate genetic experimentation. In just a few years, murine lipoproteins have been characterized, genetic variants of apolipoproteins identified,16 17 and genetic variation in susceptibility to atherosclerosis among inbred mouse strains demonstrated. Several mouse models manifesting lesions resembling human transplant arteriosclerosis have been developed18 19 20 21 22 and have helped address specific interventional issues and the mechanism of the disease. The main limitation to the mouse model is the technical complexity, which requires an extended training period for the operator to become proficient. Therefore, we decided to establish a simplified mouse model of transplant arteriosclerosis that uses a cuff technique.
In the current study, we performed allografts of carotid arteries or aortas to carotid arteries between C57BL/6J and BALB/c mice, evaluated the role of ICAM-1 in the development of allograft lesions by using ICAM-1–deficient mice, and demonstrated that ICAM-1 plays an important role in the pathogenesis of transplant arteriosclerosis.
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Methods |
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Mice and Artery Allograft Procedure
All animal experiments were performed according to protocols approved by the Institutional Committee for Use and Care of Laboratory Animals. Artery transplantation was performed between BALB/c (H2d) and C57BL/6J (H2b) (Charles River Laboratories, Sulzfeld, Germany) or ICAM-1–deficient mice of the C57BL/6J strain, which were purchased from The Jackson Laboratory, Bar Harbor, Me. Three genotypes of ICAM-1 –/–, +/-, and +/+ mice were identified by using The Jackson Laboratory’s polymerase chain reaction (PCR) protocol (primers: oIMR017 5'-CTG AGC CAG CTG GAG GTC TCG-3'; oIMR018 5'-GAG CGG CAG AGC AAA AGA AGC-3'; and oIMR019 5'-AGG ACA GCA AGG GGG AGG ATT-3'). The mice were maintained on a light/dark (12/12-hour) cycle at 22°C and received food and water ad libitum.
Two types of arteries, the carotid artery and aorta, were used as organ
donors in the current study. Mice were anesthetized with
pentobarbital sodium (50 mg/kg body weight IP), and heparin (100 U in
100 µL of saline solution) was administered via the
inferior vena cava. After 3 minutes, the right common
carotid artery was exposed and removed (
1 cm). For aorta
preparation, a midline abdominal incision was made in the donor animal,
the anterior thoracic cage was opened from the diaphragm and incised
laterally to the internal mammary vessels, and the aorta was removed.
Each vessel graft was harvested carefully to avoid mechanical injury
during surgical preparation. All grafts were washed with saline
solution containing 100 U/mL heparin.
Recipients were anesthetized, and atropine sulfate (1.7 mg/kg
body weight IP) was administered to maintain the respiratory tract in
good condition. The operation was performed under a dissecting
stereomicroscope (Olympus SZH 10). The mouse was fixed in a supine
position with its neck extended. A midline incision was made on the
ventral side of the neck from the lower mandible to the sternum. The
right cleidomastoid muscle was resected. Figure 1
shows the transplantation procedure.
The right common carotid artery was mobilized free from the bifurcation
in the distal end toward the proximal end as far as possible. The
vessel was ligated with an 8-0 silk suture and dissected between the
middle ties. The proximal and distal portions of the artery were passed
through cuffs made of an autoclavable nylon tubing, 0.63 mm in
outside diameter and a 0.5-mm inside diameter (catalog No.
800/200/100/200; Portex Ltd). The cuff length was 1 mm, with a
1-mm "handle" or extension. The vessel, together with the handle,
was fixed by microhemostat clamps (4 mm long, Martin). The suture
at the end of the artery was removed, and a segment of the artery was
everted over the cuff body and fixed to the cuff with an 8-0 silk
suture. Another portion of the artery was similarly prepared (Figure 1A).
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The arterial segment was implanted between the 2 ends of
the carotid artery by sleeving the ends of the arterial
segment over the artery cuff and ligating them together with the 8-0
suture (Figures 1B
to 1D). The vascular clamps were removed and
evidence of pulsations was sought in both the grafted and native
vessels. If there were vigorous pulsations in the transplanted vessel,
the skin incision was closed with a 6-0 interrupted suture. About 30
minutes were needed to perform the whole operation, and the
ischemia time of artery segments was between 5 and 10
minutes
For histological analysis, perfusion was performed as described previously.23 In brief, mice were anesthetized, perfused with 0.9% NaCl solution via cardiac puncture to the left ventricle, and subsequently perfusion-fixed with 4% phosphate-buffered formaldehyde (pH 7.2) for 2 and 5 minutes, respectively. The grafts were harvested at 1 day or 1, 2, 4, and 6 weeks postoperatively (6 to 8 randomly chosen mice at each time point) by cutting the transplanted segments from the native vessels at the cuff end. Samples were fixed with 4% phosphate-buffered formaldehyde at 4°C for 24 hours. For frozen section preparation, mice were killed by cervical dislocation, and arterial grafts were harvested, immediately frozen in LN2, and stored at -80°C.
Histological and Morphometric Analyses
The grafts were processed for routine histology and embedded in
paraffin. Sections (4 µm) began at the center of the graft and
were stained with hematoxylin and eosin (HE) for
histological evaluation. Sections were reviewed for
quantification of the lesions with a BX60 microscope (Olympus Optical
Co, Ltd) equipped with a Sony 3CCD camera and television monitor. The
intima was defined as the region between the lumen and the internal
elastic lamina. The media was defined as the region between the
internal and external elastic laminas. By using a transmission scanning
microscope (Zeiss LSM 510, Zeiss) equipped with a 488-nm argon-ion
laser and Plan Neofluar 10x/0.3 oculars and interfaced to the program
Start LSM 510, images were first scanned, saved, and then overlaid with
different lines to trace the lumen, the internal elastic lamina, and
the external elastic lamina. The (neo)intimal area was determined by
subtracting the area of the lumen from the area enclosed by the
internal elastic lamina. The medial area was determined by subtracting
the area enclosed by the internal elastic lamina from the area of the
external elastic lamina. Six to 8 cross sections were obtained by
selecting the first of every 3 sections from each graft. Areas were
measured and recorded in square micrometers. In the
statistical analyses, the individual values for the area from
each animal at each time point (1, 2, 4, and 6 weeks) were averaged.
From these data, the neointima/media ratios were
obtained.
The number of cell nuclei was counted in the intima, media, and adventitia region by using an eyepiece equipped with a measuring grid (WH10X-H with SQ10/10, Olympus). The area of the eyepiece grid measured 1x1 mm and consisted of 100 units (10x10). For counting the number of cell nuclei, a magnification of 400x was used. In each section, the number of cell nuclei was determined in 10 units measuring 0.025 mmx0.025 mm each (area of 625 µm2 per unit), and the mean values were recorded for each histological section.
Immunofluorescence Staining
The procedure used for immunofluorescent staining was
similar to that described previously.24 In brief, serial
5-µm-thick frozen sections were labeled with rat monoclonal
antibodies against mouse MAC-1 (CD11b/18) leukocytes, CD4, and CD8
(PharMingen) or a mouse monoclonal antibody against -actin
conjugated with FITC (Sigma Chemical Co). Sections were then labeled
with rabbit anti-rat antibodies conjugated with FITC. After being
washed, the sections were examined with a fluorescence
microscope.
En Face Immunofluorescence
The procedure used in this experiment was similar to that
described previously.25 In short, each
arterial graft segment was cut longitudinally, mounted
endothelium side up on a glass slide (2.6x7.5 cm), and
air-dried for 1 to 2 hours at room temperature. The segments were fixed
in cold acetone (-20°C) for 10 minutes and rinsed in PBS. The
segments were then incubated with appropriately diluted rat monoclonal
antibody to MAC-1 for 30 minutes. After being washed in PBS, they were
incubated with FITC-labeled rabbit anti-rat immunoglobulin for 30
minutes and washed again 3 times with a change of PBS every 30 minutes.
All incubations were performed at room temperature. Finally, sections
were mounted in gelvatol/PBS and examined with the fluorescence
microscope.
Statistical Analysis
Statistical analyses were performed on a Macintosh
computer (GraphPad Prism 2.0 program) using ANOVA and Student’s
t test. Results are given as mean±SEM. The level for
statistical significance was set at values of P<0.05 for
all comparisons.
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Results |
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Figure 1
demonstrates the procedure of an
arterial segment transplanted to the carotid artery by
using the cuff technique, by which 140 arterial grafts
between C57BL/6J (H2b) and BALB/c
(H2d) mice were studied with a surgical success
rate of 90%. About half an hour was needed for performing the
operation, and ischemia time of graft segments was between 5
and 10 minutes. The vessel grafts were harvested at various time points
and histologically examined, and 5 grafts per group
were analyzed by immunofluorescence.
Carotid or Aortic Allograft Arteriosclerosis
Representative histological
sections of control carotid artery and arterial grafts are
shown in Figure 2. In the control artery,
4 to 5 layers of cells formed the intima and media, whereas the
adventitia was composed of connective tissues (Figure 2a).
Obviously, cell infiltration into the media and neointimal
formation were found in the allografts of C57BL/6J to BALB/c mice 2
weeks postoperatively (Figure 2b). By 4 weeks,
neointimal area was enlarged, resulting in a significant
stenosis of the vessel (Figure 2c).
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Using the same technique, we performed experiments with aortic segments
as grafts transplanted (C57BL/6J to BALB/c mice) to carotid arteries.
Figure 3 shows the results, which
indicate neointimal development in grafted arteries.
Likewise, stenosis of the vessels was observed by 4 weeks, and
the lumen was completely occluded (2 of 6 grafts) by 6 weeks after
transplantation (Figures 3e and 3f). To statistically
analyze lesion development, Figure 4 summarizes data of
neointima thickness measured microscopically.
Neointima appeared as early as 2 weeks after surgery,
although no significant difference was found compared with controls.
The neointimal thickness increased in 2-week artery grafts
and significantly progressed throughout all time points. During lesion
development, 4- and 10-fold increases in neointimal area
were found in 4- and 6-week grafts, respectively (Figure 4). In
addition, experiments with aortic isografts implanted to carotid
arteries in BALB/c mice were also performed, but no
neointimal lesions were observed 4 weeks after
transplantation (data not shown).
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Reduced Lesions in ICAM-1 –/– Allografted Arteries
The arterial wall of ICAM-1 –/– and wild-type
mice is similarly composed of intima, media, 4 or 5 layers of smooth
muscle cells and adventitia, and a small amount of connective tissue
(Figures 3a and 3d). Transplanted arteries donated by
wild-type C57BL/6J mice at 4 and 6 weeks (Figures 3e and 3f)
showed neointimal hyperplasia, ie, thickening of the vessel
wall up to 20 layers of cells. Interestingly, neointimal
lesions of arterial grafts from ICAM-1 –/– to BALB/c mice
showed a marked reduction at 4 as well as 6 weeks (Figures 3b and 3c). Figure 4 summarizes data of neointima
thickness and the ratio of neointima and media as measured
microscopically. A significant difference in lesion size of
transplanted arteries derived from ICAM-1 –/– and wild-type mice was
found (P<0.01). Neointimal lesion area of
artery grafts donated by ICAM-1 –/– mice was reduced 50% to 60%
compared with wild-type controls (Figure 4).
To study any effects of ICAM-1 expression of other cells or organs on
neointimal formation, an artery transplantation from BALB/c
to ICAM-1–knockout mice was performed. When aortic segments donated by
BALB/c mice were allografted to carotid arteries of ICAM-1 –/– or
ICAM-1 +/+ mice, neointimal lesions developed in both types
of mice (Figure 5). The lumen
stenosis and the neointimal area of grafted
arteries were different between ICAM-1 +/+ and ICAM-1 –/– recipients,
although no statistical significance was found.
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Decreased Leukocyte Adhesion to and Infiltration in Artery Grafts
of ICAM-1 –/– Mice
We previously adopted the vessel en face
immunofluorescence method25 for
semiquantifying cells adhering to the endothelium of
vascular segments. This method was generally useful in clarifying the
kinetics and phenotypes of cells adhering to the vascular
endothelial surface in vivo. Nonspecific reactivity was
minimal in the negative control labeled with normal rat serum (Figure 6a), and cells adhering to the
endothelial surface were positively stained with a rat
monoclonal antibody recognizing MAC-1–positive leukocytes (CD11b/18;
Figure 6b through 6d). A large number of MAC-1–positive
leukocytes were observed adhering to the endothelium of
transplanted artery segments of wild-type mice 1 day after operation
(Figure 6c), whereas cells adherent to the surface of artery
grafts from ICAM-1 –/– mice were much less profound (Figure 6d). Occasionally, MAC-1–positive stained cells were also seen
on the surface of freshly harvested artery segments (Figure 6b).
These results indicate that leukocyte adhesion to the
endothelium is one of the earliest cellular events in
transplant-accelerated arteriosclerosis.
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To assay whether cell tethering to the endothelium is
followed by transmigration and localization in the grafted artery,
cells infiltrated in 1-week grafts were evaluated. Figures 7a through 7d shows the
histological data that indicate cell infiltration.
Normal arterial walls of both ICAM-1 +/+ (Figure 7a)
and ICAM-1 –/– (Figure 7c) mice exhibited a similar structure,
ie, 4 to 5 layers of cells, whereas increased numbers of cells in
1-week artery grafts were evident. Interestingly, abundant cell
infiltration into the vessel wall donated by ICAM-1+/+ mice (Figure 7b) was observed compared with that from ICAM-1 –/– mice
(Figure 7d). When cell nuclei in the intima and media of control
and grafted vessels were counted in 10 units of the measuring grid,
total cell numbers of the grafts from ICAM-1 +/+ mice were
significantly higher than those from ICAM-1 –/– mice 1 week
postoperatively (Figure 7e).
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Using immunofluorescent techniques, we found that these
infiltrated cells in 1-week artery grafts were MAC-1–positive,
CD4-positive cells (Figure 8) and rare
CD8-positive T cells. MAC-1–positive cells were
monocytes/macrophages, natural killer cells, and
granulocytes. The majority of infiltrating cells in the grafts was
mononuclear cells, ie, monocytes/macrophages. Abundant
infiltration of these positive cells was found in the media of 1-week
artery grafts from ICAM-1 +/+ mice (Figure 8b), whereas only low
numbers of MAC-1–positive cells were seen in the artery grafts of
ICAM-1 –/– mice (Figure 8c). MAC-1–positive
monocytes/macrophages were also detected at the luminal surface
1 week after grafting in both ICAM-1 –/– and +/+ mice (Figures 8b and 8c), but these positive cells were rarely seen in the
normal vessel (Figure 8a). Figure 8d summarizes the mean
data obtained from 5 mice per group, indicating a significant
difference in the numbers of MAC-1–positive cells, but not
CD4-positive T cells, that infiltrated into the artery grafts from
ICAM-1 +/+ and ICAM-1 –/– mice.
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SMC Proliferation in Neointima
A previous study had demonstrated the presence of abundant
SMCs in neointimal lesions of transplanted arteries. In the
current study, immunofluorescence staining with
monoclonal antibodies against -actin on frozen sections also
demonstrated the presence of abundant SMCs in neointima
lesions 4 and 6 weeks after surgery (Figure 9). No positive staining was seen in
artery segments labeled with normal rat serum as a negative control
(Figure 9a). Strong staining was observed in sections of the
grafted artery from ICAM-1+/+ mice 4 weeks postoperatively (Figure 9b). Importantly, the number of positively stained SMCs was
markedly reduced in artery grafts donated by ICAM-1 –/– mice at 4
weeks (Figure 9c). These observations indicate that the major
cells in neointimal lesions 4 weeks or thereafter
postoperatively are SMCs.
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Discussion |
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Murine genetic models in which genes are overexpressed, deleted, or mutated have been developed. These mouse models have considerable advantages over other animal systems in that they overcome the need to administer factors or their inhibitors. In the current report, we establish and characterize a new model for the study of transplant-accelerated arteriosclerosis in mice. When used with allografts from mice subjected to targeted gene deletion, the model provide a powerful tool for dissecting the relative contributions of such genes, eg, ICAM-1, in the development of neointimal hyperplasia. When used with allografts that can be treated ex vivo with drugs or gene transfer, the vessel wall allows easy penetration by small molecules and plasmids from the adventitia side. By using this model, we believe that significant progress in understanding the pathogenesis and treatment of transplant-accelerated arteriosclerosis may be seen in the near future.
Shi et al18 established the first mouse model of
transplant arteriosclerosis by end-to-side suturing
of carotid artery segments to carotid arteries, and Koulack et
al26 developed aortic transplantation to the infrarenal
aorta by an end-to-end anastomosis. These mouse models have proved to
be useful tools in studying the pathogenesis of transplant
arteriosclerosis.19 27 28 29 30 31 In the
current study, we describe a simplified mouse model of transplant
arteriosclerosis that has several advantages:
First, the operative procedure is simple and easy to learn. The vast
majority of investigators could perform the surgery after a short
period of training time. Second, the traumatic and ischemic
injuries to the grafts are minimal. Half an hour is needed to perform
the whole operation by our trained surgeon, and the ischemic
time of artery segments is between 5 and 10 minutes. Third, the success
rate of surgery is higher since the operation is performed in the neck
region and takes a shorter time. Finally, neointimal lesion
development in the transplanted vessel segments of both carotid and
aortic arteries is comparable to that of transplanted arteries by other
methods.22 31 For instance, Shi et al31
demonstrated that neointimal lesions were reduced by 52%
in allograft arteries donated by ICAM-1 –/– mice, whereas our
observations indicate a 60% reduction in neointimal
lesions of ICAM-1 –/– arteries (Figure 4). Taken together, our
mouse model has proved to be a simple and powerful tool for studying
the pathogenesis of the disease and therapeutic intervention for
transplant arteriosclerosis.
In addition, 2 types of donor organs, carotid arteries and aortas, have been used to transplant into carotid arteries in the current experiments. Although accelerated arteriosclerosis develops in both allografts, the following differences exist: (1) From a technical point of view, aortic segments are relatively easy to be sleeved over the cuff, which can be used for grafting the vessel donated by smaller mice. Since the body weight of some mutant mice is less than one third of the wild-type controls, the diameter of the carotid artery is not big enough to anastomose with the end of the cuff. (2) The intimal lesion area of aortic grafts is mostly larger than that of carotid arteries. Possibly, mononuclear cells adhere more easily to the uneven surface of the aortic intima owing to the existence of many intercostal arterial branches. (3) Intimal lesions of carotid artery allografts are of relatively uniform size and shape because they lack branches. Therefore, both donor organs can be used for transplantation, depending on the donor size and the skill of the investigator.
Although the importance of ICAM-1 in mediating cell adhesion to the
endothelium has been established, little is known about
the role of ICAM-1 expressed in allografted vessels during the
development of transplant
arteriosclerosis.31 In the current
study, we demonstrated that one of the earliest cellular events is
leukocyte adhesion to the surface of allografted vessels, which
declines in ICAM-1 –/– allografts. Obviously, these adherent cells
are destined to infiltrate into the vessel wall, because the numbers of
MAC-1–positive leukocytes in ICAM-1 +/+ artery grafts are
significantly higher than those in ICAM-1 –/– mice (Figure 8).
Therefore, ICAM-1 adhesion molecules expressed in allografted vessels
are critical in mediating leukocyte adhesion and infiltration. There is
evidence that infiltrated (activated)
monocytes/macrophages produce a large number of
cytokines and proteinases,32 of which matrix
metalloproteinases play an important role in initiating SMC
migration.33 34 We observed that a hallmark of grafted
vessels from ICAM-1 +/+ mice at the early stages is macrophage
infiltration into the media and SMC accumulation in the intima at the
late stage. Both macrophage infiltration and SMC accumulation
are reduced in ICAM-1 –/– allografted vessels. We postulate that
infiltrated macrophages may be critical in initiating SMC
migration from the media to the intima, where they proliferate to form
arteriosclerotic lesions.
Several reports provide evidence that ICAM-1 is highly expressed
on endothelial cells as well as SMCs of transplanted
arteries, cardiac allografts, and atherosclerotic
lesions.35 36 37 38 39 40 41 42 We have also observed that
neointimal SMCs express ICAM-1 (data not shown). Given the
fact that SMCs of ICAM-1 –/– mice do not express ICAM-1, which is
correlated with reduced neointimal lesions, we postulate a
crucial role for ICAM-1 expression on SMCs in the development of
intimal hyperplasia. The interaction of MAC-1 and ICAM-1 expressed on
SMCs may initiate the intracellular signaling necessary for
cytokine secretion by monocytes/macrophages. Support
for this notion comes from the fact that macrophage
inflammatory protein-1 production was induced in monocytes
cultured on ICAM-1–coated plates.43 In addition, it has
been reported that expression of ICAM-1 on SMCs may be relevant to the
phenotypic change of SMCs,44 which is considered to be
essential to their migration and proliferation in the pathogenesis of
atherosclerosis.45 Therefore, the
interaction of MAC-1 with ICAM-1 on SMCs might also play a part in the
pathogenesis of transplant arteriosclerosis.
In summary, we have established a new mouse model of transplant arteriosclerosis, which has proved to be a powerful tool for studying the mechanisms of disease development. We have provided solid evidence that neointimal lesions are reduced up to 60% in artery allografts donated by ICAM-1 –/– mice. If other adhesion molecules, including vascular cell adhesion molecule and selectins, which have also been identified as being expressed in the arterial wall,46 47 48 49 could be "knocked out" or inhibited in parallel, transplant arteriosclerosis might be prevented or retarded. Thus, further study on the role of adhesion molecules could provide valuable information for the therapeutic intervention of transplant arteriosclerosis.
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Acknowledgments |
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This work was supported by grant P12847-MED and P-13099-BIO from the Austrian Science Fund and P7917 from the Jubiläumsfonds of the Austrian National Bank. We thank A. Jenewein for excellent technical assistance and T. Öttl for the preparation of photographs.
Received June 2, 1999; accepted July 28, 1999.
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