(Circulation. 2000;101:2290.)
© 2000 American Heart Association, Inc.
Basic Science Reports |
Prominent Role of P-Selectin in the Development of Advanced Atherosclerosis in ApoE-Deficient Mice
From the Center for Blood Research (Z.M.D., A.A.B., D.D.W.), Department of Pathology (Z.M.D., D.D.W.), Harvard Medical School, Boston, Mass.
Correspondence to Denisa D. Wagner, PhD, Center for Blood Research, Harvard Medical School, 800 Huntington Ave, Boston, MA 02115.
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Abstract |
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Background—Adhesive interactions between leukocytes and endothelial cells are characteristic of the development of atherosclerotic lesions, but the receptors involved remain to be defined. P-selectin is an adhesion receptor expressed on activated endothelial cells or platelets and was shown to be involved in fatty streak formation in LDL receptor–deficient mice on an atherogenic diet. The main purpose of this study is to examine the role of P-selectin in the spontaneous development of advanced atherosclerosis in apoE-deficient mice.
Methods and Results—We intercrossed P-selectin–deficient mice with mice lacking apoE and compared lesion development in apoE-deficient mice with P-selectin (apoE-/- P+/+) and without P-selectin (apoE-/- P-/-) that were fed normal mouse chow. At 4 months of age, apoE-/- P-/- mice had 3.5-fold smaller aortic sinus lesions than apoE-/- P+/+ mice. These were limited to fatty streaks in the apoE-/- P-/- mice, whereas 70% of apoE-/- P+/+ lesions contained smooth muscle cells. Significantly more of the aortic sinus circumference was covered by lesions in the apoE-/- P+/+ animals. The P-selectin genotype affected macrophage recruitment, because twice as many mononuclear cells were present in the P-selectin–positive lesions. At 15 months, the lesions progressed to the fibrous plaque stage in both genotypes and spread throughout the aorta, but this process was delayed in apoE-/- P-/- mice. In the aortic sinus, the lesions of the apoE-/- P-/- mice were 2.6-fold smaller and less calcified.
Conclusions—P-selectin appears to be a key adhesion receptor mediating leukocyte recruitment into lesions and promoting advanced atherosclerosis in apoE-deficient mice.
Key Words: selectins • atherosclerosis • lipoproteins • leukocytes
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Introduction |
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Atherosclerosis is an inflammatory disease of large and medium-size arteries. Lesion development involves leukocyte, especially monocyte, adhesion to the vascular endothelium and migration into the intima, where it becomes a macrophage or foam cell.1 Experimental evidence shows that in human and animal atherosclerosis, one of the observed changes in the endothelium is an increase in the expression of leukocyte adhesion receptors, such as P-selectin, E-selectin, vascular cell adhesion molecule-1 (VCAM-1), and intercellular adhesion molecule-1.2 However, the role of these adhesion molecules in mediating monocyte infiltration and promoting the late stages of fibrous plaque lesion development in atherogenesis has not been clearly defined.
P-selectin is a member of the selectin family of adhesion receptors and is expressed on activated endothelium and platelets.3 It binds to monocytes, neutrophils, T cells, and platelets and mediates the first step (rolling) in leukocyte extravasation.4 5 Soluble P-selectin is proposed to be a marker of endothelial dysfunction in hypercholesterolemic patients.6 P-selectin polymorphism is reported to be associated with human atherosclerosis.7 To test the role of P-selectin in the formation of atherosclerotic lesions, we have intercrossed P-selectin–deficient mice with mice lacking the LDL receptor (LDLR-/-), which are susceptible to atherosclerosis when fed an atherogenic diet. After 8 weeks on this diet, male mice lacking both LDLR and P-selectin showed a 2-fold reduction in fatty streak lesion size in the aortic sinus. Such reduction was not observed in the females. At 37 weeks on the diet, the lesions in the LDLR-/- animals progressed to the fibrous plaque stage, and lesion size and distribution were no longer dependent on P-selectin.8 The atherogenic "Paigen" diet9 10 includes an artificially high cholesterol content (10 to 20 times that of a Western-type diet) plus the unnatural dietary constituent cholic acid. This diet could produce unintended pathophysiological processes in these animals. We have shown that it induced leukocyte rolling in the LDLR-/- mice.8 E-selectin, which has an overlapping function with P-selectin,3 could be upregulated during the proinflammatory state of mice on this diet. Furthermore, expression of VCAM-1 on the endothelium in experimental atherosclerosis has been found to be positively correlated to cholesterol levels.11 VCAM-1 has also been found to mediate leukocyte arrest as well as rolling.12 13 Therefore, the atherogenic diet could reduce the role of P-selectin in both early and late stages of lesion development in the LDLR-/- mice because of an excessive induction of other adhesion molecules with similar functions.
For this reason, we were interested in evaluating the role of P-selectin in atherosclerosis in an animal model that develops atherosclerosis spontaneously. ApoE-/- mice have elevated cholesterol levels (300 to 600 mg/dL)10 and develop fibrous plaque lesions with features characteristic of those seen in humans.14 We therefore compared the lesion development in apoE-deficient mice with and without P-selectin maintained on normal chow diet.
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Methods |
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Mice, Genotype Analysis, and Diet
P-selectin–deficient (P-/-) mice15 were backcrossed 4 times to C57BL/6J. ApoE-deficient (apoE-/-) mice backcrossed 8 times to C57BL/6J were purchased from Jackson Laboratory.16 P-/- mice were interbred with apoE-/- mice. Littermates from the F2 generation of this intercross were used to establish apoE-/- P+/+ matings and apoE-/- P-/- matings used in our study. Polymerase chain reaction analysis was performed for genotyping for P-selectin17 and cholesterol measurement for apoE.16 Mice were maintained on a normal chow diet containing 5% fat (wt/wt) (Prolab 3000; PMI Feeds).
Cholesterol Determination
Mice were fasted overnight, and blood was collected through the
retro-orbital venous plexus in polypropylene tubes containing EDTA.
Cholesterol in total plasma was determined with a kit
(352-50, Sigma).
Blood Counts
Blood was obtained as above. Complete blood counts were
determined with an automatic cell counter (Coulter).
Quantification of Aortic Sinus Lesions
The sections containing aortic sinus were prepared as
described,8 with modifications. The heart and attached
aorta were fixed in 4% paraformaldehyde for 30 minutes
before being embedded in OCT compound. Four consecutive 10-µm
sections were collected for each slide. Ten slides were made from each
animal. Odd-numbered slides were stained with oil red O and
hematoxylin, counterstained with light green, and examined for lesion
size and calcium deposits. Even-numbered slides were kept for
immunohistochemical analysis. The area of the lesion was
measured with a Leica Q500 MC image analysis program (Leica
Inc). Values reported represent the mean lesion area from 5
sections for each animal.
Quantification of Lesions in Entire Aorta
The procedure for quantification of the lesions was described in
Reference 18 .
Histological Analysis
Calcium deposits were identified by the hematoxylin
stain.18 19 Mononuclear cells/macrophages were
identified by their cellular and nuclear appearance and counted on
hematoxylin-stained sections.
Immunohistochemical Analysis
To identify macrophages in spleens, frozen sections
10 µm thick were fixed in cold acetone for 5 minutes.
Endogenous peroxidase activity was blocked with 3%
hydrogen peroxide for 5 minutes and then incubated with a
biotin-conjugated rat anti-mouse F4/80 antibody (BioSource
International). Antibodies were visualized by an avidin/biotin
peroxidase–linked detection system (Vector Laboratories). To quantify
smooth muscle cells, 2 sections on even-numbered slides for each mouse
were stained with a mouse monoclonal antibody against human 
-actin
directly coupled to horseradish peroxidase (No. U7033, dilution 1:2;
Dako Corp).18 The -actin–positive area was measured
with an ocular micrometer.
Statistical Analysis
Data are presented as mean±SEM. Student’s t
test was performed with software for MacIntosh. Because a relatively
small number of animals were available for the 15-month group, the
Mann-Whitney U test (2-tailed) was selected. Pearson
r product-moment correlations were calculated.
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Results |
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Comparison of ApoE-Deficient Mice With and Without P-Selectin
We bred P-/- mice with apoE-/- mice to generate apoE-/- P+/+ and apoE-/- P-/- mice. At 4 months of age on normal chow, male and female apoE-/- P+/+ and apoE-/- P-/- mice had similar body weights and comparable blood levels of cholesterol (Table
).
However, beyond 
12 months of age, most male
apoE-/- P-/- mice
started to lose body weight (wasting), whereas this did not happen to
female apoE-/- P-/-
mice. Therefore, at 15 months of age, apoE-/-
P-/- male mice had significantly lower body
weight (34.7±1.8 versus 49.3±4.6 g, n=7, P=0.0086) and
plasma cholesterol (407±26 versus 756±51 mg/dL,
P=0.0008) than apoE-/-
P+/+ control male mice.
ApoE-/- P-/- female
mice maintained body weights and cholesterol levels similar
to those of apoE-/- P+/+
control female mice at that age (Table). Because body weight and
cholesterol are significant factors affecting
atherosclerotic lesion development,20 21 we excluded
the older male mice from this study. Unlike mice deficient in
P-selectin only,22 apoE-/-
P-/- mice consistently displayed
splenomegaly. Splenic weights, normalized to body weights, from
apoE-/- P-/- and
apoE-/- P+/+ mice were
increased 2-fold at 4 months of age and 4-fold at 15 months of age
in favor of the apoE-/-
P-/- mice (Table). Microscopic
examination of spleen sections from apoE-/-
P-/- mice disclosed hyperplasia with an
increased amount of white pulp and expanded red pulp (not shown), with
larger infiltrates of F4/80-positive macrophages (Figure 1). Weights of livers,
peripheral blood total leukocyte counts, and platelet
counts were comparable between apoE-/-
P+/+ and apoE-/-
P-/- mice.
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Reduction in Lesion Size in ApoE-Deficient Mice Without
P-Selectin
The extent of the lesions in the aortic sinus has been shown to
correlate well with that of the entire aorta in
apoE-/- mice.23 Mean lesion areas
per cross section were measured in oil red O–stained tissue sections.
At 4 months of age on normal chow, apoE-/-
P-/- male mice had lesions 3.6-fold smaller
than the apoE-/- P+/+
males (0.039±0.001 versus 0.142±0.036 mm2,
n=7 to 14, P=0.002), and apoE-/-
P-/- female mice had lesions 3.4-fold smaller
than the apoE-/- P+/+
females (0.077±0.013 versus 0.258±0.03
mm2, n=12 or 13, P<0.0001). The
combined data from males and females are shown in Figure 2. In addition, at this time point, 74%
of the aortic sinus circumference in apoE-/-
P+/+ mice was covered by lesions, compared with
30% found in apoE-/-
P-/- mice (P<0.0001, n=19 to 27).
Thus, apoE-/- P-/- mice
had more lesion-free aortic sinus surface as well as smaller lesions
than apoE-/- P+/+
mice.
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With increasing age, lesions grew in size in both
apoE-/- P+/+ and
apoE-/- P-/- mice.
However, a significant difference in lesion size between the 2
genotypes persisted at 15 months, with 2.6-fold smaller lesions
in apoE-/- P-/- mice
and no overlap in lesion size between the 2 groups (Figure 2
).
Among all the 15-month-old animals combined, the
cholesterol levels did not correlate with their lesion size
(r=0.2, P=NS), indicating that the differences in
lesion size were not linked to minor cholesterol variations
but rather to the P-selectin genotype. At this time point, the
lesions spread throughout the entire aorta. The percentage of surface
area occupied by the lesions was determined from the subclavian branch
to the iliac bifurcation and was significantly higher in
apoE-/- P+/+ aortas than
apoE-/- P-/- aortas
(Figure 3).
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Comparison of Lesion Composition in ApoE-Deficient Mice With and
Without P-Selectin
Through histological analysis and
immunohistochemical staining, we characterized the stages of the
atherosclerotic lesions in the apoE-/-
P+/+ and apoE-/-
P-/- mice. At 4 months of age, 70% (13/19) of
apoE-/- P+/+ mice had
already developed fibrofatty or early fibrous plaque lesions containing
smooth muscle cells (Figure 4, A and C),
and 30% (6/19) remained at the fatty streak stage. In contrast, all
(27/27) of the apoE-/-
P-/- mice developed only a fatty streak lesion
at this stage (Figure 4B). No smooth muscle cells were found in
these lesions by -actin staining (Figure 4D). The fibrofatty
lesions of apoE-/- P+/+
mice contained a mixture of foam cells and spindle-shaped cells,
presumably smooth muscle cells (Figure 4A). The early fibrous
plaques encompassed small necrotic cores, together with a few foam
cells that were covered by a well-formed fibrous cap containing
-actin–positive smooth muscle cells (Figure 4C). We stained
aortic sinus sections of the 4-month-old
apoE-/- mice with hematoxylin and counted all
mononuclear cells in the lesions. We found that the lesions in
P+/+ mice had twice as many mononuclear cells
(173±27 per section, n=14) as the lesions of
P-/- mice (88±15 per section, n=19). This
difference was significant (P<0.006) and showed that the
absence of P-selectin inhibited the recruitment of
monocytes/macrophages to the lesion.
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Furthermore, at 15 months of age, the fibrous plaques in
apoE-/- P+/+ appeared
more advanced, with larger necrotic cores, more abundant fibrous
tissue, and more calcification (Figure 4E) than in
apoE-/- P-/- mice.
Although the lesions in the latter also progressed to the fibrous
plaque stage at this age, they were less calcified and contained less
fibrous tissue (Figure 4F). The calcification frequencies of
fibrous plaque lesions were 6 of 8 in the
apoE-/- P+/+ and 1 of 6
in the apoE-/- P-/-
mice. The area of the lesion covered by -actin–positive smooth
muscle cells tended to be less in apoE-/-
P-/- mice than in
apoE-/- P+/+ mice (6.5%
versus 11.8%; P=0.058).
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Discussion |
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P-selectin is located in the membranes of
-granules in
platelets and the Weibel-Palade bodies in
endothelial cells.24 On stimulation,
P-selectin is rapidly translocated to the cell surface, where it
mediates adhesion to carbohydrate ligand(s) on
leukocytes.25 P-selectin is involved in many acute
inflammatory conditions mediated by neutrophils,26 27 and
because it regulates monocyte and T-lymphocyte emigration from the
blood stream,22 28 it may also be involved in chronic
inflammatory conditions, such as atherosclerosis.
P-selectin has been detected on the surface of the
endothelium in all stages of atherosclerotic lesion
development in humans and in animal models of
atherosclerosis.2 Many atherogenic
factors, such as oxidized LDL and inflammatory cytokines,
induce P-selectin expression.29 30 31 Ramos et
al32 demonstrated that anti–P-selectin antibodies inhibit
monocyte rolling on endothelium of the carotid artery
isolated from apoE-deficient mice. However, these studies could not
distinguish whether the increased expression of P-selectin is a cause
or a consequence of the atherosclerotic process. The availability of
mice with a deficiency in P-selectin makes it possible to address these
issues directly. Using C57BL/6 mice fed the atherogenic diet, Nageh and
colleagues33 observed a 50% to 70% reduction in fatty
streak size in the P-/- mice. Unfortunately,
because normal mice are resistant to the development of fibrous
plaque,10 this study could not evaluate the role of
P-selectin in the more advanced stages of lesion development. As
described in the introduction, using LDLR-deficient mice fed the
atherogenic diet, we found that the absence of P-selectin partially
reduces fatty streak formation but does not affect the more advanced
stages of atherosclerosis.8 The
present study is the first to demonstrate that P-selectin is also
important for lesion growth at the fibrous plaque stage. If we consider the many differences between the LDLR-deficient and apoE-deficient mouse atherosclerosis models,10 it is not surprising to find that P-selectin plays a more prominent role in the apoE-/- mice than in the LDLR-/- mice. On a normal chow diet, LDLR-/- mice reach a total plasma cholesterol of 200 mg/dL, which is only 2-fold higher than that of wild-type mice, and do not develop any gross aortic lesions up to 13 months of age.34 35 In contrast, apoE-/- mice on chow food achieve cholesterol levels of 400 to 600 mg/dL and develop fatty streak and fibrofatty (intermediate lesion) and fibrous plaque starting at 2.5, 4, and 5 months of age, respectively.14 The increased lipoproteins, which may serve as atherogenic stimuli and inducers of adhesion molecules on endothelium in LDLR-/- mice, are the IDL and LDL, whereas the lipoproteins in apoE-/- mice are chylomicron remnants and VLDL.10 To induce lesion development in LDLR-/- mice, a high-fat/high-cholesterol diet such as the Paigen atherogenic diet9 is required.35 This diet, which contains cholic acid, can stimulate an inflammatory response and could induce expression of adhesion molecules such as E-selectin. Accordingly, we have found that the size of the fibrous plaque formed in the aortic sinus of LDLR-deficient mice on the atherogenic diet for 37 weeks is identical in the presence and absence of P-selectin but is reduced significantly, by 40%, when both P- and E-selectin are absent.18 On the basis of our present study in the apoE-deficient mouse, in which absence of P-selectin alone inhibited advanced lesion progression, we suspect that the profile and relative importance of adhesion molecules that promote lesion development in different animal models of atherosclerosis and different subgroups of human patients with this disease can vary.
One mechanism that may be responsible for decreased lesion progression
in the absence of P-selectin in apoE-deficient mice is the inhibition
of leukocyte, especially monocyte, infiltration into the lesions.
Indeed, we observed 2-fold fewer mononuclear cells in the lesions of
4-month-old apoE-deficient mice lacking P-selectin. Monocyte
attachment to endothelial cells of the aortic sinus in
apoE-deficient mice was observed before obvious fatty streak
formation.36 Macrophages loaded with
fluorescent microspheres injected
intravenously into 40-week-old apoE-deficient mice adhered
to all stages of atherosclerotic plaques, including mature calcified
fibrous lesions.37 Macrophages in the
subendothelium may at first protect against
atherosclerosis but later also contribute to lesion
progression.1 38 It is also possible that the absence of
P-selectin modifies systemic immune response in the animals, which
could affect lesion progression. Atherosclerotic lesions, even in the
apoE-deficient mouse model, are infiltrated with T helper (Th) 1
cells38 secreting proinflammatory cytokines. Th1
cells but not the anti-inflammatory Th2 cells can bind to P-selectin
and thus efficiently enter Th1-dominated inflammatory
sites.39 P-selectin deficiency could cause an imbalance in
the recruitment of the Th1 versus Th2 cells, which could have a
significant effect on the lesion progression and maturation. Indeed, we
observed that the absence of P-selectin caused a delay in lesion
maturation, with a smaller proportion of smooth muscle cells and
reduced calcification (Figure 4). Platelets may also
contribute to lesion development,1 and P-selectin mediates
platelet interaction with both endothelial cells
and leukocytes.4 5
In this study, we unexpectedly found that apoE-/- P-/- mice had splenomegaly, with an increased number of macrophages. The cause of the large spleen in these animals is not clear. It is not due to infections, because these mice had a normal blood leukocyte count. We hypothesize that the monocytes, which were prevented from migrating into the intima in the absence of P-selectin, homed to spleen, became macrophages, and finally caused hyperplasia in this organ. Macrophage homing to spleen in apoE-/- mice appears not to depend on leukocyte adhesion molecules.37 Excessive, chronic accumulation of macrophages in spleen may cause these cells to become transformed. We had found on autopsy that 2 of 4 apoE-/- P-/- mice developed histiocytic sarcoma (a malignant tumor of macrophages) before 1 year of age. In addition, P-selectin has a protective role in experimental glomerulonephritis.40 Consistent with this finding, we also found on autopsy that 3 of 4 apoE-/- P-/- mice spontaneously developed glomerulonephritis. These diseases are the likely cause of wasting observed in the apoE-/- P-/- mice, especially males, leading to a higher mortality at 1 year (35% in apoE-/- P-/- mice versus <10% in apoE-/- P+/+ or mice deficient in P-selectin only). Although anti–P-selectin therapy may be a powerful tool in inhibiting atherosclerotic lesion progression, possible side effects should be considered in evaluating its therapeutic potential.
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Acknowledgments |
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This study was supported by NIH NHLBI grant HL-53756 (Dr Wagner) and fellowship HL-09869 (Dr Dong). A.A. Brown was supported by a grant from the Genetics Institute. The care of the experimental mice was in accordance with the guidelines of the ACUC of the Center for Blood Research. We are grateful to Lesley Cowan and Maria Economopoulos for help in preparing the manuscript and to Dr Sheldon Wagner for statistical analysis.
Received August 24, 1999; revision received December 6, 1999; accepted December 17, 1999.
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References |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
1. Ross R. The pathogenesis of atherosclerosis: a perspective for the 1990s. Nature. 1993;362:801–809.[Medline] [Order article via Infotrieve]
2. Dong ZM, Wagner DD. Leukocyte-endothelium adhesion molecules in atherosclerosis. J Lab Clin Med.. 1998;132:369–375.[Medline] [Order article via Infotrieve]
3. Tedder TF, Steeber DA, Chen A, Engel P. The selectins: vascular adhesion molecules. FASEB J. 1995;9:866–873.[Abstract]
4. Frenette PS, Wagner DD. Insights into selectin function from knockout mice. Thromb Haemost.. 1997;78:60–64.[Medline] [Order article via Infotrieve]
5.
Frenette PS, Johnson RC, Hynes RO, Wagner DD.
Platelets roll on stimulated endothelium in vivo:
an interaction mediated by endothelial P-selectin.
Proc Natl Acad Sci U S A. 1995;92:7450–7454.
6.
Davi G, Romano M, Mezzetti A, Procopio A, Iacobelli S,
Antidormi T, Bucciarelli T, Alessandrini P, Cuccurullo F, Bon GB.
Increased levels of soluble P-selectin in
hypercholesterolemic patients. Circulation. 1998;97:953–957.
7.
Herrmann SM, Ricard S, Nicaud V, Mallet C, Evans A,
Ruidavets JB, Arveiler D, Luc G, Cambien F. The P-selectin gene is
highly polymorphic: reduced frequency of the Pro715 allele
carriers in patients with myocardial infarction. Hum Mol
Genet. 1998;7:1277–1284.
8. Johnson RC, Chapman SM, Dong ZM, Ordovas JM, Mayadas TN, Herz J, Hynes RO, Schaefer EJ, Wagner DD. Absence of P-selectin delays fatty streak formation in mice. J Clin Invest. 1997;99:1037–1043.[Medline] [Order article via Infotrieve]
9. Nishina PM, Verstuyft J, Paigen B. Synthetic low and high fat diets for the study of atherosclerosis in the mouse. J Lipid Res. 1990;31:859–869.[Abstract]
10. Breslow JL. Mouse models of atherosclerosis. Science. 1996;272:685–688.[Abstract]
11.
Nakashima Y, Raines EW, Plump AS, Breslow JL, Ross R.
Upregulation of VCAM-1 and ICAM-1 at
atherosclerosis-prone sites on the
endothelium in the apoE-deficient mouse.
Arterioscler Thromb Vasc Biol. 1998;18:842–851.
12.
Berlin C, Bargatze RF, Campbell JJ, von Andrian UH,
Szabo MC, Hasslen SR, Nelson RD, Berg EL, Erlandsen SL, Butcher
EC. 4 integrins mediate lymphocyte attachment and rolling under
physiologic flow. Cell. 1995;80:413–422.[Medline]
[Order article via Infotrieve]
13.
Alon R, Kassner PD, Carr MW, Finger EB, Hemler ME,
Springer TA. The integrin VLA-4 supports tethering and rolling in flow
on VCAM-1. J Cell Biol.. 1995;128:1243–1253.
14.
Nakashima Y, Plump AS, Raines EW, Breslow JL, Ross R.
ApoE-deficient mice develop lesions of all phases of
atherosclerosis throughout the arterial
tree. Arterioscler Thromb. 1994;14:133–140.
15. Mayadas TN, Johnson RC, Rayburn H, Hynes RO, Wagner DD. Leukocyte rolling and extravasation are severely compromised in P-selectin-deficient mice. Cell. 1993;74:541–554.[Medline] [Order article via Infotrieve]
16.
Zhang S, Reddick RL, Piedrahita JA, Maeda N.
Spontaneous hypercholesterolemia and
arterial lesions in mice lacking apolipoprotein E.
Science. 1992;258:468–471.
17. Frenette PS, Mayadas TN, Rayburn H, Hynes RO, Wagner DD. Susceptibility to infection and altered hematopoiesis in mice deficient in both P- and E-selectins. Cell.. 1996;84:563–574.[Medline] [Order article via Infotrieve]
18. Dong ZM, Chapman SM, Brown AA, Frenette PS, Hynes RO, Wagner DD. The combined role of P- and E-selectins in atherosclerosis. J Clin Invest.. 1998;102:145–152.[Medline] [Order article via Infotrieve]
19.
Qiao JH, Xie PZ, Fishbein MC, Kreuzer J, Drake TA,
Demer LL, Lusis AJ. Pathology of atheromatous
lesions in inbred and genetically engineered mice: genetic
determination of arterial calcification. Arterioscler
Thromb. 1994;14:1480–1497.
20. Nishina PM, Naggert JK, Verstuyft J, Paigen B. Atherosclerosis in genetically obese mice: the mutants obese, diabetes, fat, tubby, and lethal yellow. Metabolism. 1994;43:554–558.[Medline] [Order article via Infotrieve]
21.
Smith JD, Trogan E, Ginsberg M, Grigaux C, Tian J,
Miyata M. Decreased atherosclerosis in mice deficient
in both macrophage colony-stimulating factor (op) and
apolipoprotein E. Proc Natl Acad Sci U S A. 1995;92:8264–8268.
22.
Johnson RC, Mayadas TN, Frenette PS, Mebius RE,
Subramaniam M, Lacasce A, Hynes RO, Wagner DD. Blood cell dynamics in
P-selectin-deficient mice. Blood. 1995;86:1106–1114.
23. Tangirala RK, Rubin ER, Palinski W. Quantitation of atherosclerosis in murine models: correlation between lesions in the aortic origin and in the entire aorta, and differences in the extent of lesions between sexes in LDL receptor-deficient and apolipoprotein E-deficient mice. J Lipid Res. 1995;36:2320–2328.[Abstract]
24. Wagner DD. The Weibel-Palade body: the storage granule for von Willebrand factor and P-selectin. Thromb Haemost. 1993;70:105–110.[Medline] [Order article via Infotrieve]
25.
McEver RP, Moore KL, Cummings RD. Leukocyte trafficking
mediated by selectin- carbohydrate interactions. J Biol
Chem. 1995;270:11025–11028.
26. Bevilacqua MP, Nelson RM, Mannori G, Cecconi O. Endothelial-leukocyte adhesion molecules in human disease. Annu Rev Med. 1994;45:361–378.[Medline] [Order article via Infotrieve]
27. Jang Y, Lincoff AM, Plow EF. Cell adhesion molecules in coronary artery disease. J Am Coll Cardiol.. 1994;24:1591–1601.[Abstract]
28.
Subramaniam M, Saffaripour S, Watson SR, Mayadas TN,
Hynes RO, Wagner DD. Reduced recruitment of inflammatory cells in a
contact hypersensitivity response in P- selectin-deficient mice.
J Exp Med. 1995;181:2277–2282.
29.
Mehta A, Yang B, Khan S, Hendricks JB, Stephen C, Mehta
JL. Oxidized low-density lipoproteins facilitate leukocyte adhesion to
aortic intima without affecting endothelium- dependent
relaxation: role of P-selectin. Arterioscler Thromb Vasc
Biol.. 1995;15:2076–2083.
30.
Vora DK, Fang ZT, Liva SM, Tyner TR, Parhami F, Watson
AD, Drake TA, Territo MC, Berliner JA. Induction of P-selectin by
oxidized lipoproteins: separate effects of synthesis and surface
expression. Circ Res. 1997;80:810–818.
31. Gimbrone MA Jr, Nagel T, Topper JN. Biomechanical activation: an emerging paradigm in endothelial adhesion biology. J Clin Invest. 1997;100(suppl):S61–S65.
32.
Ramos CL, Huo Y, Jung U, Ghosh S, Manka DR, Sarembock
IJ, Ley K. Direct demonstration of P-selectin– and VCAM-1–dependent
mononuclear cell rolling in early atherosclerotic lesions of
apolipoprotein E–deficient mice. Circ Res. 1999;84:1237–1244.
33.
Nageh MF, Sandberg ET, Marotti KR, Lin AH, Melchior EP,
Bullard DC, Beaudet AL. Deficiency of inflammatory cell adhesion
molecules protects against atherosclerosis in mice.
Arterioscler Thromb Vasc Biol. 1997;17:1517–1520.
34. Ishibashi S, Brown MS, Goldstein JL, Gerard RD, Hammer RE, Herz J. Hypercholesterolemia in low density lipoprotein receptor knockout mice and its reversal by adenovirus-mediated gene delivery. J Clin Invest. 1993;92:883–893.
35. Ishibashi S, Goldstein JL, Brown MS, Herz J, Burns DK. Massive xanthomatosis and atherosclerosis in cholesterol-fed low density lipoprotein receptor-negative mice. J Clin Invest.. 1994;93:1885–1893.
36.
Tamminen M, Mottino G, Qiao JH, Breslow JL, Frank JS.
Ultrastructure of early lipid accumulation in apoE-deficient mice.
Arterioscler Thromb Vasc Biol. 1999;19:847–853.
37.
Patel SS, Thiagarajan R, Willerson JT, Yeh ETH.
Inhibition of 4 integrin and ICAM-1 markedly attenuate
macrophage homing to atherosclerotic plaques in
apo-E–deficient mice. Circulation. 1998;97:75–81.
38. Zhou X, Paulsson G, Stemme S, Hannsson GK. Hypercholesterolemia is associated with a T helper (Th) 1/Th2 switch of the autoimmune response in atherosclerotic apo E-knockout mice. J Clin Invest. 1998;101:1717–1725.[Medline] [Order article via Infotrieve]
39. Austrup F, Vestweber D, Borges E, Lohning M, Brauer R, Herz U, Renz H, Hallmann R, Scheffold A, Radbruch A, Hamann A. P- and E-selectin mediate recruitment of T-helper-1 but not T-helper-2 cells into inflamed tissues. Nature. 1997;385:81–83.[Medline] [Order article via Infotrieve]
40. Rosenkranz AR, Mendrick DL, Cotran RS, Mayadas TN. P-selectin deficiency exacerbates experimental glomerulonephritis: a protective role for endothelial P-selectin in inflammation. J Clin Invest. 1999;103:649–659.[Medline] [Order article via Infotrieve]
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