Published online before print December 1, 2003, doi: 10.1161/01.RES.0000109793.17591.4E
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
© 2004 American Heart Association, Inc.
Integrative Physiology |
Antagonism of RANTES Receptors Reduces Atherosclerotic Plaque Formation in Mice
From the Division of Cardiology (N.R.V., B.K., G.P., F. Mulhaupt, F. Mach), Foundation for Medical Research; Endocrinology and Diabetes Division (R.W.J.), Faculty of Medicine, University Hospital Geneva; and Serono Pharmaceutical Research Institute (A.E.I.P.), Geneva, Switzerland.
Correspondence to François Mach, MD, Cardiology Division, Department of Medicine, University Hospital Geneva, Foundation for Medical Research, 64 Ave Roseraie, 1211 Geneva 4, Switzerland. E-mail Francois.Mach{at}medecine.unige.ch
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Increasing evidence supports the involvement of inflammation in the early phases of atherogenesis. Recruitment of leukocytes within the vascular wall, controlled by chemokines, is an essential process in the development of this common disease. In this study, we report that blocking a chemokine pathway in vivo with the CC chemokine antagonist Met-RANTES reduces the progression of atherosclerosis in a hypercholesterolemic mouse model. The reduction of lesions was correlated with a diminution of expression of several major chemokines and chemokine receptors, a decrease in leukocyte infiltration, and an increase of collagen-rich atheroma, features associated with stable atheroma. Treatment was well tolerated and serum lipid profiles were not affected. Whereas genetically engineered mice with deletion of either a CC chemokine or its receptor have demonstrated resistance to disease, to our knowledge, this is the first demonstration that treatment with a chemokine receptor antagonist limits the progression of atherosclerosis in vivo. Thus, our findings indicate that blockade of chemokine receptor/ligand interactions might become a novel therapeutic strategy to reduce the evolution of this common disease.
Key Words: atherogenesis • leukocytes • vascular cells • chemokines • chemokine receptor antagonists
|
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Atherosclerosis is an inflammatory disease characterized by arterial lesions containing cholesterol, immune infiltrates, and fibrosis,1–3 resulting essentially from hyperlipidemia, hypertension, smoking, and diabetes.4 These cardiovascular risk factors cause endothelial dysfunction, which triggers the migration of leukocytes, mainly of monocyte/macrophage and T lymphocyte type, within the vessel wall intima area. Proinflammatory factors, such as cytokines and chemokines, released from endothelial cells (ECs), macrophages, or T cells, cause proliferation and migration of smooth muscle cells (SMCs) from the media to the intima, as well as recruitment of new immunoinflammatory cells. Within the intima, SMCs secrete extracellular matrix components, leading to the accumulation of collagen and proteoglycans, key factors implicated in plaque stability. Conversely, the secretion of matrix metalloproteinases by vascular and inflammatory cells degrades matrix components, such as collagen, gelatin, or elastin within atherosclerotic lesions.
Chemokines (chemotactic cytokines) belong to a large superfamily of low molecular weight proteins with a highly homologous 3-dimensional structure, which are divided into four groups based on the configuration of the first two cysteines.5,6 Chemokines are known to induce leukocyte migration, growth, and activation through 7 transmembrane heptahelical, G protein–coupled cell-surface receptors on target cells and regulate leukocyte trafficking during inflammation. The chemokine CCL5/RANTES (Regulated on Activated Normal T-Cell Expressed and Secreted) is a soluble chemokine of 7.8 kDa secreted by many different cell types, such as ECs, SMCs, activated T cells, macrophages, and platelets.7,8 RANTES interacts with the chemokine receptors CCR1, CCR3, and CCR5, and has been implicated in cardiac inflammatory disorders after organ transplantation or arterial injury.9,10 In addition, RANTES has been detected in plasma samples of patients suffering from cardiovascular diseases.11
When RANTES is produced recombinantly in prokaryotic cells, the initiating methionine residue is retained (Met-RANTES), resulting in a RANTES receptor antagonist with nanomolar potency, which is able to block calcium mobilization and chemotaxis,12 and induces only weak internalization of the receptor CCR513 and CCR1.14 Furthermore, it has been demonstrated that Met-RANTES do not retains high affinity for the murine chemokine receptors mCCR3.15 This antagonist has been shown to significantly reduce inflammatory responses in several animal models including crescentic glomerular nephritis,16 rheumatoid arthritis,17 airways inflammation,18 and organ transplant rejection.19 However, in a recent publication, Kuziel et al20 reported that CCR5 deficiency was not protective in the early stages of atherosclerosis in ApoE knockout mice. These intriguing results are surprising knowing that RANTES avidly binds CCR5 and that this receptor is highly expressed on the principal cell types implicated during atherogenesis such as T cells,21 macrophages,22 SMCs,23 and ECs.24
Based on the crucial role of immunoinflammatory processes during atherogenesis and the fact that RANTES may trigger monocyte/macrophages and T lymphocytes chemotaxis and activation, we decided to investigate the effect of Met-RANTES treatment on atherosclerotic plaque formation in vivo.
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Reagents
The human chemokine antagonist Met-RANTES was generated as previously described.12 For in vivo experiments, Met-RANTES was dissolved in sterile water and diluted to a concentration of 1 µg/µL (125 µmol/L) in PBS before administration. Rabbit anti-mouse CD4 (Pharmingen, clone H129.19), rabbit anti-mouse Mac-1 (Pharmingen, clone M1/70), rabbit anti-mouse RANTES (R&D, clone 53405.111), goat anti-mouse MMP9 (R&D), and rabbit anti-mouse smooth muscle myosin heavy chain (gift from Dr G. Gabbiani, Geneva, Switzerland) antibodies were used for immunohistochemical analysis. Mouse anti-human RANTES capture antibody (R&D, clone 21418) and goat anti-human RANTES detection antibody (R&D) were used for ELISA analysis. Lipids were stained with Sudan IV and collagen with Sirius red. Chemokine and chemokine receptor expressions were detected by ribonuclease protection assay (RPA) using RPA Multi-Probe Template Set (Pharmingen, probe sets mCR-5 and mCK-5b). [125I]Met-RANTES has been custom radiolabeled (Amersham). Stable CHO transfectants expressing mCCR1 and mCCR5 were provided by Dr Gerry Graham (Beatson Institute, Glasgow, UK).
Animals
As a model of in vivo atherosclerosis, 12-week-old male LDLR-/- C57BL/6J mice (Iffa Credo, Lyon, France) were fed with high-cholesterol diet (1.25% cholesterol, 0% cholate; product No. D12108, Research Diets) in conventional housing. Littermate mice were divided into three different groups: control, saline (PBS), and Met-RANTES (n=8 per group for histological analysis and n=6 per group for mRNA analysis). Treatment by Met-RANTES was administered by intraperitoneal injection (100 µg) twice a week during 14 weeks (in parallel with diet). For histological and immunohistochemical analysis, refer to Kwak et al,25 and for more details, see reference 1 in the expanded Materials and Methods in the online data supplement available at http://www.circresaha.org.
Western Blotting
For Western blotting, 100 ng of recombinant mouse RANTES (R&D) and Met-RANTES were loaded on tricine peptide separation gels and transferred on PVDF Immobilon-Psq Transfer Membrane (Millipore Laboratory Corp). After blocking step, blots were incubated for 1 hour at room temperature with Met-RANTES treated mouse serum or mouse preserum (prior treatment) diluted at 1:100 in BSA 3%. Secondary peroxidase-conjugated antibody (1:5000 in BSA 3%; Jackson ImmunoResearch) was incubated for 1 hour. As a loading control, Coomassie blue staining was performed.
ELISA
In a separate set of experiments, mice treated with Met-RANTES (n=6) were euthanized at a different time after injection. To assess the concentration of Met-RANTES within mice sera, anti-human RANTES capture antibody was used in combination with anti-human RANTES detection antibody. The protocol and antibody concentrations were used according to the manufacturer’s instructions.
Chemokine Receptor Specificity
Equilibrium competition binding assays were performed according to Van Riper at al.26 For transmigration assay details, refer to reference 2 in the expanded Materials and Methods.
[125I]Met-RANTES
Male LDLR-/- mice fed a hypercholesterolemic diet during 2 months, as well as control WT mice not fed with high-fat diet, were injected (intraperitoneally) with 2 µCi radiolabeled [125I]Met-RANTES 12 hours before euthanasia. Aortas were isolated as described before and exposed onto developing film (Kodak Biomax MS Film) for 7 days at -80°C. Mice sera and aortas were used to detect [125I]Met-RANTES concentration. CPM measured from aortas were normalized by dividing total counts of each sample by its dried weight.
RPA
Total mouse mRNA was extracted from total aorta (aortic arch and thoracoabdominal aorta) and prepared with Tri reagent (MRC Inc) according to the manufacturer’s instructions. RPA with 10 µg of mRNA per reaction were performed using commercial RPA Multi-Probe Template Set as recommended by the supplier. Signal quantitation was determined using a PhosphorImager (Molecular Dynamics) and normalized to GAPDH and L32 housekeeping genes.
Blood Analysis and Leukocyte Quantification
Hematocrit and leukocyte counts were measured, and SERA were used for measurement of cholesterol and triglycerides content.27 HDL and VLDL cholesterol fractions were measured by fast protein liquid chromatography.28 Presence of macrophages and lymphocytes within lesions was determined by quantification of immunohistochemistry for Mac-1 and CD4, respectively. Percentage positive areas were reported as areas within intima as previously described.29
Statistical Analysis
All results are expressed as mean±SEM. Differences between the groups were considered significant at P<0.05 using the Student’s t test or the Mann-Whitney U Wilcoxon sum test.
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Expression of the Chemokine RANTES Is Colocalized at Site of Inflammation With Atheroma-Associated Cells
We first analyzed the expression of the chemokine RANTES comparing nondiseased arteries and atherosclerotic vessels. Whereas expression of RANTES was not detected in normal arteries, immunohistochemistry performed on mouse aortic root showed that RANTES was highly expressed within the atheroma (Figure 1A). Staining for macrophages and T lymphocytes on consecutive sections revealed localization of RANTES with these cell-types, in particular with macrophage foam cells (Figure 1A). Similar results were obtained with human carotid atherosclerotic lesions (data not shown).
|
) and T cells on serial sections revealed localization with RANTES. Similar results were obtained in separate experiments using 5 different mice. Scale bar=20 µm. B, Concentration of Met-RANTES within mice sera at different times after injection detected by ELISA. Control (Ctrl) represents mice not treated with Met-RANTES. C, Antibodies produced by mice treated with Met-RANTES. Western blots were performed by using mice sera as first antibodies. Coomassie staining (top) and immunoblotting (bottom) were performed on transfer membranes. M-R indicates human Met-RANTES; R, recombinant mouse RANTES.
Met-RANTES Binds to Vascular Endothelium and Retains High Affinity for Murine Receptors
In order to show the localization of Met-RANTES within vascular tissue, we injected intraperitoneally LDLR-/- and wild-type (WT) mice with [125I]Met-RANTES. The radioactive labeled molecule could be found all along the aorta and highly concentrated at site of atheroma (Figure 2A) of LDLR-/- mice. In contrast, only weak signals could be detected within WT mice. Counts of the 125I isotope activity indicated that LDLR-/- mice had 2.8-fold and 2.3-fold more radioactive Met-RANTES compared with WT mice within serum and aorta, respectively (online Table 1, in the online data supplement). In addition, we performed receptor-binding assays and showed that Met-RANTES retains high-affinity binding for the murine receptors CCR1 and CCR5 (Figures 2B and 2C). [125I]-mMIP-1
could be displaced by hRANTES and Met-RANTES from mCCR1 with an IC50 of 89 and 79 nmol/L, respectively. Similarly, [125I]-mMIP-1 could be displaced by hRANTES and Met-RANTES from mCCR5 with an IC50 of 18 and 61 nmol/L, respectively. In order to investigate whether Met-RANTES could bind CCR2, one of the most important receptor implicated in atherosclerosis, we performed transmigration assays using macrophages isolated from peritoneal exudates of ApoE-/- and ApoE-/- CCR2-/--deficient mice. Met-RANTES inhibited significantly cell migration induced by RANTES, but no difference was observed in chemotaxis for macrophages obtained from either mouse strains (Figure 2D). In another set of experiments using ApoE-/- mouse macrophages, Met-RANTES could not block chemotaxis induced by MCP-1 (Figure 2E), even at high concentrations of Met-RANTES. Similar results were obtained using THP1 cells (data not shown).
|
Met-RANTES Influences Atherosclerotic Plaque Development and Composition
To determine the possible beneficial effect of treatment with the chemokine antagonist Met-RANTES in vivo, we analyzed the development and the quality of atherosclerotic plaques within LDLR-/- mice aortas fed a high-cholesterol diet.30 We randomly assigned these mice to either no treatment, saline, or Met-RANTES treatment over a period of 14 weeks. As an internal control, we measured by ELISA the concentration of Met-RANTES within mice sera at different times after injection with an anti-human RANTES capture antibody completely lacking cross-reactivity with endogenous murine RANTES. We showed that Met-RANTES could be rapidly detected in mice SERA 2 hours after injection and reached a maximal concentration of about 2 ng/mL (250 pmol/L) at 12 hours, and then decreased slowly to reach similar concentrations at 40 hours as those observed after 2 hours (Figure 1B). Thus, these results indicate that, by injecting our mice twice a week, we are dealing with low and nonsaturating concentrations of Met-RANTES. Administration of the human Met-RANTES protein into mice indeed induced antibody production. However, by using SERA from mice treated by Met-RANTES as the first antibody, we demonstrated that the detection of the antagonist was specific for the human molecule, as antibodies could not bind to murine recombinant RANTES (Figure 1C). In addition, Met-RANTES treatment did not affect RANTES protein expression within mouse aorta compared with controls (data not shown). We also verified that Met-RANTES did not alter other essential control parameters, and no statistical differences were detected within the three different groups for weight, hematocrit, blood leukocyte count, and lipid profile modifications (Table). The distribution in lipoprotein fractions (VLDL and HDL) was determined by fast protein liquid chromatography (FPLC) and showed no difference between controls and Met-RANTES–treated animals (Figure 3). Furthermore, neither liver nor kidney toxicity was found and none of our mice died during treatment nor showed unhealthy behavior (data not shown). Control and saline-treated mice exhibited large and extended atherosclerotic lesions compared with Met-RANTES–treated animals. Examination of aortic roots (Figures 4A through 4C) as well as thoracoabdominal aortas (Figures 4D through 4F) stained with Sudan IV for lipids showed a significant decrease in plaque formation in mice treated by Met-RANTES. The results showed that the extent of lesions was significantly decreased within Met-RANTES–treated mice compared with controls, with 43% reduction for aortic roots and 58% reduction for thoracoabdominal aortas (Figure 4G).
|
|
, saline; 
, Met-RANTES; and 
, control.
|
To investigate in more detail the composition of the atherosclerotic plaques after Met-RANTES treatment, we compared the quality and the cellularity of atherosclerotic lesions between groups. Macrophage foam cells and T lymphocytes were significantly more abundant within tissues isolated from controls (Figures 5A and 5B) compared with Met-RANTES–treated mice (Figures 5C and 5D). Indeed, quantification of immunohistochemistry for Mac-1 and CD4 present within aortic lesions confirmed a reduction of macrophages and lymphocytes in Met-RANTES–treated animals, by 43% and 83%, respectively (Figure 5E). Analysis of SMC quantity into the neointima showed relevant differences between each treatment group: SMCs were more abundant within a thicker fibrous cap covering atherosclerotic lesions in Met-RANTES–treated mice compared with controls (Figures 5F, 5G, and 5L). Detailed analysis of plaque components also revealed that Met-RANTES treatment increased interstitial collagen content, a product of SMCs, at the shoulder of the lesion compared with a very poor concentration observed within control atheroma (Figures 5H, 5I, and 5M). To further explore these intriguing results, we performed staining for matrix metalloproteinases (MMPs), secreted mainly by macrophages and SMCs, and observed a lower expression of MMP-9 within atherosclerotic lesions in Met-RANTES–treated mice compared with control lesions (Figures 5J, 5K, and 5N). Thus, we can hypothesize that the reduced degradation of collagen might be principally due to the difference in MMP secretion from the lower number of macrophages, but might also result from a lower activation state of SMCs within Met-RANTES–treated mice compared with controls. Knowing that vulnerability and rupture of atherosclerotic lesion are dependent on the thickness and number of SMCs within the fibrous cap, we can speculate an additional beneficial effect of Met-RANTES on the stability of atherosclerotic plaques.
|
Expression of Proinflammatory Molecules
We then analyzed the expression of certain chemokine receptors and their proinflammatory chemokine ligands known to be implicated during atherogenesis. We observed a highly significant reduction of mRNA expression for the chemokine receptors CCR2 (-86%) and CCR5 (-87%) in mice treated with Met-RANTES (Figures 6A and 6B), thus confirming a reduction in inflammation within vascular atherosclerotic tissue. In a second set of experiments, we analyzed the expression of certain chemokines within the same vascular tissues. Surprisingly, we observed no significant variation of the expression of the chemokines RANTES, CCL2/MCP-1 (monocyte chemoattractant protein 1) and XCL11/lymphotactin (Ltn) between the different treated groups. Nevertheless, for the chemokines CCL11/eotaxin, CCL3/macrophage inflammatory protein 1 (MIP-1), CCL4/macrophage inflammatory protein 1ß (MIP-1ß), and macrophage inflammatory protein 2 (MIP-2), mRNAs within vascular atherosclerotic tissue were expressed at lower levels in Met-RANTES–treated animals by 35%, 40%, 40%, and 34%, respectively (Figures 6C and 6D). Data for chemokine and chemokine receptor mRNAs were normalized to GAPDH, as well as for L32 with same results. In order to investigate whether the reduced expression of these proinflammatory molecules resulted mainly from the lower number of inflammatory cells present within atherosclerotic lesions, we normalized our data for macrophage content (Figure 7). Tested for statistically significant differences, our results demonstrated that reduced expression of the receptors CCR5 and CCR2 were still significant after this normalization procedure. However, expression of the chemokine eotaxin, MIP-1, MIP-1ß, and MIP-2 normalized to macrophage content did not result in any longer significant differences between control and Met-RANTES–treated animals.
|
|
|
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Inflammatory infiltrates, mainly composed of T-lymphocytes and mononuclear phagocytes, are recruited by a network of chemokines and chemokine receptors highly expressed by vascular cells within atherosclerotic lesions.5 The chemokines RANTES and MCP-1 are potent activators and chemoattractant signals of inflammatory and vascular cells by binding to CCR1, CCR2, and CCR5 receptors.31–34 The role of MCP-1 has previously been established by both MCP-1– and CCR2-deficient mice.35,36 Several experiments have described the expression and implication of RANTES in animal models of inflammation by treatment with the chemokine receptor antagonist Met-RANTES.16–19 Thus, we decided to investigate the effect of inhibiting RANTES actions on the development of atherosclerosis in a mouse model fed with high-cholesterol diet using Met-RANTES.
First, we demonstrated that RANTES is highly expressed within atheroma, that Met-RANTES had significant antiinflammatory effects even though it was only detectable in the serum at picomolar concentrations, and that antibodies produced against Met-RANTES do not cross-react with the murine RANTES. Detection of Met-RANTES in murine plasma 40 hours after injection indicates that chemokines are protected from degradation in vivo. This stabilization of chemokines may result from their immobilization on cell surface glycosaminoglycans (GAGs).37 The detection of Met-RANTES in murine serum would then be explained by a slow release mechanism from cell surface GAGs into the circulation. This mechanism could explain the long-lasting activity of Met-RANTES in vivo. In addition, we demonstrated that Met-RANTES could bind to vascular tissue, especially at atherogenic sites, and that Met-RANTES retains high affinity for murine CCR1 and CCR5 and do not bind to CCR2.
Treatment with Met-RANTES reduced the extent of atherosclerosis lesions analyzed on aortic roots and thoracoabdominal aortas. These effects could be linked to a considerable decrease in leukocyte infiltration, to an increase of atherosclerotic lesion stability, as well as to a reduction in expression of certain chemokines and chemokine receptors. Levels of CCR5 and CCR2 receptors were significantly decreased within vascular tissues isolated from Met-RANTES–treated mice. The role of CCR2, highly expressed on macrophages, but also on SMCs7 and T lymphocytes,38 especially on Th1 T cells (the principal T lymphocyte type detected within atheroma), is of a major impact for inflammatory processes during atherogenesis. Detection of CCR5 within atherosclerotic lesions confirms a previous report.39 Our results could demonstrate a possible importance of the receptor CCR5 in the development of atherosclerosis, which is in contrast with the recent observations of Kuziel and colleges20 where it has been reported that CCR5 deficiency was not protective in the early stages of atherosclerosis in ApoE-/- mice. These differences with our study might include the possibility that chemokines contribute differentially to the development of atherosclerosis in one model (LDLR-/-) but not in the other (ApoE-/-). However, these observations might more likely result from genetic variations because the generated CCR5-/- 129/Ola mice used in their study was only twice backcrossed with ApoE-/- C57BL/6 mice. Furthermore, Kuziel and colleges described effects of CCR5-/- mice only on very early atherosclerotic lesions. It would have been interesting to see whether deletion of CCR5 may alter advanced atherosclerotic plaque formation. In addition, this MIP-1/MIP-1ß/RANTES receptor, expressed on T cells, macrophages, SMCs, and ECs, has been reported to be implicated in the activation and chemotaxis of different cell types. MIP-1 has been shown, by in vitro studies, to chemoattract many different leukocyte types including T cells, monocytes, and neutrophils, to be expressed at sites of inflammation and to activate ECs.40 In animal models, neutralizing antibodies to MIP-1 have been shown to decrease inflammation by limiting monocyte and lymphocyte recruitment.40 In SMC culture, MIP-1ß induces a significant increase of intracellular calcium concentration, which could be blocked by an antibody to CCR5.23 Thus, CCR5 may play a role in the recruitment and activation of leukocytes as well as of vascular cells, and the blockade of CCR1 and CCR5 by Met-RANTES would prevent leukocyte migration into lesion. However, the reduction of CCR2 and CCR5 may be secondary to the reduced macrophage content within lesions. Indeed, our results also indicate that the reduced expression of these receptors might be due to the decreased activation state of vascular cells, and not to macrophage content within atherosclerotic lesions. Indeed, expression of these chemokine receptors per macrophage still showed significant differences between control and Met-RANTES–treated animals, thus indicating that other cells, such as SMCs and ECs might be involved.
Interestingly, regulation of chemokines showed different variation patterns. No differences were detected for the chemokines RANTES and MCP-1. Because Met-RANTES significantly decreases the number of infiltrating leukocytes, we assumed that the unaltered high transcription levels of RANTES and MCP-1 chemokine mRNAs result mainly from vascular SMCs and ECs that are activated by hypercholesterolemia. We hypothesize that the degree of activation of SMCs and ECs within the vessel wall of atherosclerotic lesion of Met-RANTES–treated mice may compensate for the loss of RANTES and MCP-1 mRNA from the decreased number of leukocytes. In contrast, significant decreases were observed for MIP-1, MIP-1ß, MIP-2, and eotaxin mRNA within vascular tissues isolated from Met-RANTES–treated mice. Nevertheless, because of the nonstatistical differences observed between each treatment group, higher expression of the chemokines eotaxin, MIP-1, MIP-1ß, and MIP-2 within control mice lesions may result only from macrophage content reflecting regulatory loops affected by Met-RANTES. Indeed, it has been reported that MIP-1 and MIP-1ß are secreted by a variety of cells including macrophages, neutrophils, and fibroblasts, but not, as far as we know, by SMCs and ECs.23 Mouse MIP-2, secreted by mononuclear cells,41 may serve the same chemotactic function as CXCL8/IL-8 in human, because IL-8 does not exist in mice.42 MIP-2 has also been shown to play a key role in neutrophil recruitment in the development of inflammation and tissue injury,43 indicating that Met-RANTES may also play an essential role in the activation and chemotactic function of leukocytes at the early stages of atherogenesis. Expression of the chemokine eotaxin, known to be secreted by many different cell type including vascular and inflammatory cells,44 seems in our study to be mainly secreted from macrophages because no significant differences were obtained by macrophage normalization. Thus, the reduction of leukocyte infiltration into the neointima might explain the lower expression of these chemokines after Met-RANTES administration. Altogether, we hypothesize that in our model of atherosclerosis, Met-RANTES may reduce the activation of the endothelium and SMCs, and also acts by limiting the recruitment of monocyte-macrophages and T lymphocytes into the vascular wall, resulting in slowing down the inflammatory processes of atherosclerosis. The reduction of macrophages within lesions may also lead to the observed striking decrease of matrix metalloproteinases leading to more stable lesions that contain more collagen.
In conclusion, blocking RANTES signaling in vivo with the CC chemokine antagonist Met-RANTES influences the development of atherosclerotic lesions and also regulates several vascular-cell functions related to acute manifestations of atherosclerosis. These findings highlight the involvement of CCR1 and/or CCR5, in addition to CCR2, as novel targets for the treatment of atherogenesis.
|
Acknowledgments |
|---|
This work was supported by grants from the Swiss National Science Foundation (No. 3200-065121.01/1 to François Mach, No. 3100-067777.02 to Brenda Kwak, No. 3100-64788.01/1 to Richard James), and by a grant from the Foundation for Medical Research (Geneva) to Niels Veillard.
|
Footnotes |
|---|
Original received May 20, 2003; revision received November 19, 2003; accepted November 19, 2003.
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
1. Libby P. Inflammation in atherosclerosis. Nature. 2002; 420: 868–874.[CrossRef][Medline] [Order article via Infotrieve]
2. Hansson GK. Inflammation and immune response in atherosclerosis. Curr Atheroscler Rep. 1999; 1: 150–155.[Medline] [Order article via Infotrieve]
3. Glass CK, Witztum JL. Atherosclerosis. the road ahead. Cell. 2001; 104: 503–516.[CrossRef][Medline] [Order article via Infotrieve]
4. Lusis AJ. Atherosclerosis. Nature. 2000; 407: 233–241.[CrossRef][Medline] [Order article via Infotrieve]
5. Luster AD. Chemokines: chemotactic cytokines that mediate inflammation. N Engl J Med. 1998; 338: 436–445.
6. Veillard NR, Mach F. Statins: the new aspirin? Cell Mol Life Sci. 2002; 59: 1771–1786.[CrossRef][Medline] [Order article via Infotrieve]
7. Hayes IM, Jordan NJ, Towers S, Smith G, Paterson JR, Earnshaw JJ, Roach AG, Westwick J, Williams RJ. Human vascular smooth muscle cells express receptors for CC chemokines. Arterioscler Thromb Vasc Biol. 1998; 18: 397–403.
8. Pattison JM, Nelson PJ, Huie P, Sibley RK, Krensky AM. RANTES chemokine expression in transplant-associated accelerated atherosclerosis. J Heart Lung Transplant. 1996; 15: 1194–1199.[Medline] [Order article via Infotrieve]
9. von Hundelshausen P, Weber KS, Huo Y, Proudfoot AE, Nelson PJ, Ley K, Weber C. RANTES deposition by platelets triggers monocyte arrest on inflamed and atherosclerotic endothelium. Circulation. 2001; 103: 1772–1777.
10. Yun JJ, Fischbein MP, Laks H, Irie Y, Espejo ML, Fishbein MC, Berliner JA, Ardehali A. RANTES production during development of cardiac allograft vasculopathy. Transplantation. 2001; 71: 1649–1656.[CrossRef][Medline] [Order article via Infotrieve]
11. Parissis JT, Adamopoulos S, Venetsanou KF, Mentzikof DG, Karas SM, Kremastinos DT. Serum profiles of C-C chemokines in acute myocardial infarction: possible implication in postinfarction left ventricular remodeling. J Interferon Cytokine Res. 2002; 22: 223–229.[CrossRef][Medline] [Order article via Infotrieve]
12. Proudfoot AE, Power CA, Hoogewerf AJ, Montjovent MO, Borlat F, Offord RE, Wells TN. Extension of recombinant human RANTES by the retention of the initiating methionine produces a potent antagonist. J Biol Chem. 1996; 271: 2599–2603.
13. Mack M, Luckow B, Nelson PJ, Cihak J, Simmons G, Clapham PR, Signoret N, Marsh M, Stangassinger M, Borlat F, Wells TN, Schlondorff D, Proudfoot AE. Aminooxypentane-RANTES induces CCR5 internalization but inhibits recycling: a novel inhibitory mechanism of HIV infectivity. J Exp Med. 1998; 187: 1215–1224.
14. Solari R, Offord RE, Remy S, Aubry JP, Wells TN, Whitehorn E, Oung T, Proudfoot AE. Receptor-mediated endocytosis of CC-chemokines. J Biol Chem. 1997; 272: 9617–9620.
15. Chvatchko Y, Proudfoot AE, Buser R, Juillard P, Alouani S, Kosco-Vilbois M, Coyle AJ, Nibbs RJ, Graham G, Offord RE, Wells TN. Inhibition of airway inflammation by amino-terminally modified RANTES/CC chemokine ligand 5 analogues is not mediated through CCR3. J Immunol. 2003; 171: 5498–5506.
16. Lloyd CM, Minto AW, Dorf ME, Proudfoot A, Wells TN, Salant DJ, Gutierrez-Ramos JC. RANTES and monocyte chemoattractant protein-1 (MCP-1) play an important role in the inflammatory phase of crescentic nephritis, but only MCP-1 is involved in crescent formation and interstitial fibrosis. J Exp Med. 1997; 185: 1371–1380.
17. Plater-Zyberk C, Hoogewerf AJ, Proudfoot AE, Power CA, Wells TN. Effect of a CC chemokine receptor antagonist on collagen induced arthritis in DBA/1 mice. Immunol Lett. 1997; 57: 117–120.[CrossRef][Medline] [Order article via Infotrieve]
18. Gonzalo JA, Lloyd CM, Wen D, Albar JP, Wells TN, Proudfoot A, Martinez AC, Dorf M, Bjerke T, Coyle AJ, Gutierrez-Ramos JC. The coordinated action of CC chemokines in the lung orchestrates allergic inflammation and airway hyperresponsiveness. J Exp Med. 1998; 188: 157–167.
19. Grone HJ, Weber C, Weber KS, Grone EF, Klier CM, Wells TN, Proudfoot AE, Schlondorff D, Nelson PJ. [Reduction of acute kidney transplantation rejection by the chemokine receptor antagonist Met-RANTES]. Verh Dtsch Ges Pathol. 1999; 83: 205–211.[Medline] [Order article via Infotrieve]
20. Kuziel WA, Dawson TC, Quinones M, Garavito E, Chenaux G, Ahuja SS, Reddick RL, Maeda N. CCR5 deficiency is not protective in the early stages of atherogenesis in apoE knockout mice. Atherosclerosis. 2003; 167: 25–32.[CrossRef][Medline] [Order article via Infotrieve]
21. Sallusto F, Lenig D, Mackay CR, Lanzavecchia A. Flexible programs of chemokine receptor expression on human polarized T helper 1 and 2 lymphocytes. J Exp Med. 1998; 187: 875–883.
22. Raport CJ, Gosling J, Schweickart VL, Gray PW, Charo IF. Molecular cloning and functional characterization of a novel human CC chemokine receptor (CCR5) for RANTES, MIP-1ß, and MIP-1. J Biol Chem. 1996; 271: 17161–17166.
23. Schecter AD, Calderon TM, Berman AB, McManus CM, Fallon JT, Rossikhina M, Zhao W, Christ G, Berman JW, Taubman MB. Human vascular smooth muscle cells possess functional CCR5. J Biol Chem. 2000; 275: 5466–5471.
24. Berger O, Gan X, Gujuluva C, Burns AR, Sulur G, Stins M, Way D, Witte M, Weinand M, Said J, Kim KS, Taub D, Graves MC, Fiala M. CXC and CC chemokine receptors on coronary and brain endothelia. Mol Med. 1999; 5: 795–805.[Medline] [Order article via Infotrieve]
25. Kwak BR, Veillard N, Pelli G, Mulhaupt F, James RW, Chanson M, Mach F. Reduced connexin43 expression inhibits atherosclerotic lesion formation in low-density lipoprotein receptor-deficient mice. Circulation. 2003; 107: 1033–1039.
26. Van Riper G, Siciliano S, Fischer PA, Meurer R, Springer MS, Rosen H. Characterization and species distribution of high affinity GTP-coupled receptors for human RANTES and monocyte chemoattractant protein 1. J Exp Med. 1993; 177: 851–856.
27. James RW, Pometta D. Differences in lipoprotein subfraction composition and distribution between type I diabetic men and control subjects. Diabetes. 1990; 39: 1158–1164.[Abstract]
28. Mach F, Schonbeck U, Sukhova GK, Atkinson E, Libby P. Reduction of atherosclerosis in mice by inhibition of CD40 signalling. Nature. 1998; 394: 200–203.[CrossRef][Medline] [Order article via Infotrieve]
29. Aikawa M, Rabkin E, Sugiyama S, Voglic SJ, Fukumoto Y, Furukawa Y, Shiomi M, Schoen FJ, Libby P. An HMG-CoA reductase inhibitor, cerivastatin, suppresses growth of macrophages expressing matrix metalloproteinases and tissue factor in vivo and in vitro. Circulation. 2001; 103: 276–283.
30. Tangirala RK, Rubin EM, 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]
31. Appay V, Dunbar PR, Cerundolo V, McMichael A, Czaplewski L, Rowland-Jones S. RANTES activates antigen-specific cytotoxic T lymphocytes in a mitogen-like manner through cell surface aggregation. Int Immunol. 2000; 12: 1173–1182.
32. Kaplan AP. Chemokines, chemokine receptors and allergy. Int Arch Allergy Immunol. 2001; 124: 423–431.[CrossRef][Medline] [Order article via Infotrieve]
33. Horuk R. Chemokine receptors. Cytokine Growth Factor Rev. 2001; 12: 313–335.[CrossRef][Medline] [Order article via Infotrieve]
34. Peters W, Charo IF. Involvement of chemokine receptor 2 and its ligand, monocyte chemoattractant protein-1, in the development of atherosclerosis: lessons from knockout mice. Curr Opin Lipidol. 2001; 12: 175–180.[CrossRef][Medline] [Order article via Infotrieve]
35. Gu L, Okada Y, Clinton SK, Gerard C, Sukhova GK, Libby P, Rollins BJ. Absence of monocyte chemoattractant protein-1 reduces atherosclerosis in low density lipoprotein receptor-deficient mice. Mol Cell. 1998; 2: 275–281.[CrossRef][Medline] [Order article via Infotrieve]
36. Boring L, Gosling J, Cleary M, Charo IF. Decreased lesion formation in CCR2-/- mice reveals a role for chemokines in the initiation of atherosclerosis. Nature. 1998; 394: 894–897.[CrossRef][Medline] [Order article via Infotrieve]
37. Proudfoot AE, Handel TM, Johnson Z, Lau EK, LiWang P, Clark-Lewis I, Borlat F, Wells TN, Kosco-Vilbois MH. Glycosaminoglycan binding and oligomerization are essential for the in vivo activity of certain chemokines. Proc Natl Acad Sci U S A. 2003; 100: 1885–1890.
38. Kim CH, Rott L, Kunkel EJ, Genovese MC, Andrew DP, Wu L, Butcher EC. Rules of chemokine receptor association with T cell polarization in vivo. J Clin Invest. 2001; 108: 1331–1339.[CrossRef][Medline] [Order article via Infotrieve]
39. Dol F, Martin G, Staels B, Mares AM, Cazaubon C, Nisato D, Bidouard JP, Janiak P, Schaeffer P, Herbert JM. Angiotensin AT1 receptor antagonist irbesartan decreases lesion size, chemokine expression, and macrophage accumulation in apolipoprotein E–deficient mice. J Cardiovasc Pharmacol. 2001; 38: 395–405.[CrossRef][Medline] [Order article via Infotrieve]
40. Lee SC, Brummet ME, Shahabuddin S, Woodworth TG, Georas SN, Leifermani KM, Gilman SC, Stellato C, Gladue RP, Schleimer RP, Beck LA. Cutaneous injection of human subjects with macrophage inflammatory protein-1 induces significant recruitment of neutrophils and monocytes. J Immunol. 2000; 164: 3392–3401.
41. Weiss SJ. Tissue destruction by neutrophils. N Engl J Med. 1989; 320: 365–376.[Medline] [Order article via Infotrieve]
42. Shanley TP, Schmal H, Warner RL, Schmid E, Friedl HP, Ward PA. Requirement for C-X-C chemokines (macrophage inflammatory protein-2 and cytokine-induced neutrophil chemoattractant) in IgG immune complex-induced lung injury. J Immunol. 1997; 158: 3439–3448.[Abstract]
43. Schmal H, Shanley TP, Jones ML, Friedl HP, Ward PA. Role for macrophage inflammatory protein-2 in lipopolysaccharide-induced lung injury in rats. J Immunol. 1996; 156: 1963–1972.[Abstract]
44. Haley KJ, Lilly CM, Yang JH, Feng Y, Kennedy SP, Turi TG, Thompson JF, Sukhova GH, Libby P, Lee RT. Overexpression of eotaxin and the CCR3 receptor in human atherosclerosis: using genomic technology to identify a potential novel pathway of vascular inflammation. Circulation. 2000; 102: 2185–2189.
This article has been cited by other articles:
 |
|
 |
|
  M. Qiao, Q. Zhao, C. F. Lee, L. R. Tannock, E. J. Smart, R. G. LeBaron, C. F. Phelix, Y. Rangel, and R. Asmis Thiol Oxidative Stress Induced by Metabolic Disorders Amplifies Macrophage Chemotactic Responses and Accelerates Atherogenesis and Kidney Injury in LDL Receptor-Deficient Mice Arterioscler Thromb Vasc Biol, November 1, 2009; 29(11): 1779 - 1786. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. L.H. Muntinghe, M. Verduijn, M. W. Zuurman, D. C. Grootendorst, J. J. Carrero, A. R. Qureshi, K. Luttropp, L. Nordfors, B. Lindholm, V. Brandenburg, et al. CCR5 Deletion Protects Against Inflammation-Associated Mortality in Dialysis Patients J. Am. Soc. Nephrol., July 1, 2009; 20(7): 1641 - 1649. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Madani, K. Karastergiou, N. C. Ogston, N. Miheisi, R. Bhome, N. Haloob, G. D. Tan, F. Karpe, J. Malone-Lee, M. Hashemi, et al. RANTES release by human adipose tissue in vivo and evidence for depot-specific differences Am J Physiol Endocrinol Metab, June 1, 2009; 296(6): E1262 - E1268. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. M. van Gils, J. J. Zwaginga, and P. L. Hordijk Molecular and functional interactions among monocytes, platelets, and endothelial cells and their relevance for cardiovascular diseases J. Leukoc. Biol., February 1, 2009; 85(2): 195 - 204. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. S. Bae, J. H. Lee, S. H. Choi, S. Kim, F. Almazan, J. L. Witztum, and Y. I. Miller Macrophages Generate Reactive Oxygen Species in Response to Minimally Oxidized Low-Density Lipoprotein: Toll-Like Receptor 4- and Spleen Tyrosine Kinase-Dependent Activation of NADPH Oxidase 2 Circ. Res., January 30, 2009; 104(2): 210 - 218. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Shah, N. Mehta, and M. P. Reilly Adipose Inflammation, Insulin Resistance, and Cardiovascular Disease JPEN J Parenter Enteral Nutr, November 1, 2008; 32(6): 638 - 644. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Zernecke, E. Shagdarsuren, and C. Weber Chemokines in Atherosclerosis: An Update Arterioscler Thromb Vasc Biol, November 1, 2008; 28(11): 1897 - 1908. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Z. Rocha, E. J. Folco, G. Sukhova, K. Shimizu, I. Gotsman, A. H. Vernon, and P. Libby Interferon-{gamma}, a Th1 Cytokine, Regulates Fat Inflammation: A Role for Adaptive Immunity in Obesity Circ. Res., August 29, 2008; 103(5): 467 - 476. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. R. Afzal, S. Kiechl, Y. P. Daryani, A. Weerasinghe, Y. Zhang, M. Reindl, A. Mayr, S. Weger, Q. Xu, and J. Willeit Common CCR5-del32 Frameshift Mutation Associated With Serum Levels of Inflammatory Markers and Cardiovascular Disease Risk in the Bruneck Population Stroke, July 1, 2008; 39(7): 1972 - 1978. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Braunersreuther, S. Steffens, C. Arnaud, G. Pelli, F. Burger, A. Proudfoot, and F. Mach A Novel RANTES Antagonist Prevents Progression of Established Atherosclerotic Lesions in Mice Arterioscler Thromb Vasc Biol, June 1, 2008; 28(6): 1090 - 1096. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. Saederup, L. Chan, S. A. Lira, and I. F. Charo Fractalkine Deficiency Markedly Reduces Macrophage Accumulation and Atherosclerotic Lesion Formation in CCR2-/- Mice: Evidence for Independent Chemokine Functions in Atherogenesis Circulation, April 1, 2008; 117(13): 1642 - 1648. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Combadiere, S. Potteaux, M. Rodero, T. Simon, A. Pezard, B. Esposito, R. Merval, A. Proudfoot, A. Tedgui, and Z. Mallat Combined Inhibition of CCL2, CX3CR1, and CCR5 Abrogates Ly6Chi and Ly6Clo Monocytosis and Almost Abolishes Atherosclerosis in Hypercholesterolemic Mice Circulation, April 1, 2008; 117(13): 1649 - 1657. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Plat, A. Jellema, J. Ramakers, and R. P. Mensink Weight Loss, but Not Fish Oil Consumption, Improves Fasting and Postprandial Serum Lipids, Markers of Endothelial Function, and Inflammatory Signatures in Moderately Obese Men J. Nutr., December 1, 2007; 137(12): 2635 - 2640. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Krohn, U. Raffetseder, I. Bot, A. Zernecke, E. Shagdarsuren, E. A. Liehn, P. J. van Santbrink, P. J. Nelson, E. A. Biessen, P. R. Mertens, et al. Y-Box Binding Protein-1 Controls CC Chemokine Ligand-5 (CCL5) Expression in Smooth Muscle Cells and Contributes to Neointima Formation in Atherosclerosis-Prone Mice Circulation, October 16, 2007; 116(16): 1812 - 1820. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Thalmann and C. A. Meier Local adipose tissue depots as cardiovascular risk factors Cardiovasc Res, September 1, 2007; 75(4): 690 - 701. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Barlic and P. M. Murphy Chemokine regulation of atherosclerosis J. Leukoc. Biol., August 1, 2007; 82(2): 226 - 236. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 O. L. Volger, J. O. Fledderus, N. Kisters, R. D. Fontijn, P. D. Moerland, J. Kuiper, T. J. van Berkel, A.-P. J.J. Bijnens, M. J.A.P. Daemen, H. Pannekoek, et al. Distinctive Expression of Chemokines and Transforming Growth Factor-{beta} Signaling in Human Arterial Endothelium during Atherosclerosis Am. J. Pathol., July 1, 2007; 171(1): 326 - 337. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Cavusoglu, C. Eng, V. Chopra, L. T. Clark, D. J. Pinsky, and J. D. Marmur Low Plasma RANTES Levels Are an Independent Predictor of Cardiac Mortality in Patients Referred for Coronary Angiography Arterioscler Thromb Vasc Biol, April 1, 2007; 27(4): 929 - 935. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. M. Matter and C. Handschin RANTES (Regulated on Activation, Normal T Cell Expressed and Secreted), Inflammation, Obesity, and the Metabolic Syndrome Circulation, February 27, 2007; 115(8): 946 - 948. [Full Text] [PDF]
|
|
|
|
|
|
V. Braunersreuther, A. Zernecke, C. Arnaud, E. A. Liehn, S. Steffens, E. Shagdarsuren, K. Bidzhekov, F. Burger, G. Pelli, B. Luckow, et al. Ccr5 But Not Ccr1 Deficiency Reduces Development of Diet-Induced Atherosclerosis in Mice Arterioscler Thromb Vasc Biol, February 1, 2007; 27(2): 373 - 379. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. von Hundelshausen and C. Weber Platelets as Immune Cells: Bridging Inflammation and Cardiovascular Disease Circ. Res., January 5, 2007; 100(1): 27 - 40. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Hernandez-Vargas, G. Ortiz-Munoz, O. Lopez-Franco, Y. Suzuki, J. Gallego-Delgado, G. Sanjuan, A. Lazaro, V. Lopez-Parra, L. Ortega, J. Egido, et al. Fc{gamma} Receptor Deficiency Confers Protection Against Atherosclerosis in Apolipoprotein E Knockout Mice Circ. Res., November 24, 2006; 99(11): 1188 - 1196. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Potteaux, C. Combadiere, B. Esposito, C. Lecureuil, H. Ait-Oufella, R. Merval, P. Ardouin, A. Tedgui, and Z. Mallat Role of Bone Marrow-Derived CC-Chemokine Receptor 5 in the Development of Atherosclerosis of Low-Density Lipoprotein Receptor Knockout Mice Arterioscler Thromb Vasc Biol, August 1, 2006; 26(8): 1858 - 1863. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A.P.J.J. Bijnens, E. Lutgens, T. Ayoubi, J. Kuiper, A.J. Horrevoets, and M.J.A.P. Daemen Genome-Wide Expression Studies of Atherosclerosis: Critical Issues in Methodology, Analysis, Interpretation of Transcriptomics Data Arterioscler Thromb Vasc Biol, June 1, 2006; 26(6): 1226 - 1235. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. V. d'Uscio and Z. S. Katusic Increased vascular biosynthesis of tetrahydrobiopterin in apolipoprotein E-deficient mice Am J Physiol Heart Circ Physiol, June 1, 2006; 290(6): H2466 - H2471. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Zernecke, E. A. Liehn, J.-L. Gao, W. A. Kuziel, P. M. Murphy, and C. Weber Deficiency in CCR5 but not CCR1 protects against neointima formation in atherosclerosis-prone mice: involvement of IL-10 Blood, June 1, 2006; 107(11): 4240 - 4243. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Mateo, Y. Naim Abu Nabah, M. Abu Taha, M. Mata, M. Cerda-Nicolas, A. E. I. Proudfoot, R. A. K. Stahl, A. C. Issekutz, J. Cortijo, E. J. Morcillo, et al. Angiotensin II-Induced Mononuclear Leukocyte Interactions with Arteriolar and Venular Endothelium Are Mediated by the Release of Different CC Chemokines J. Immunol., May 1, 2006; 176(9): 5577 - 5586. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Tedgui and Z. Mallat Cytokines in Atherosclerosis: Pathogenic and Regulatory Pathways Physiol Rev, April 1, 2006; 86(2): 515 - 581. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. B. Holven, J. K. Damas, A. Yndestad, T. Waehre, T. Ueland, B. Halvorsen, L. Heggelund, W. J. Sandberg, A. G. Semb, S. S. Froland, et al. Chemokines in Children With Heterozygous Familiar Hypercholesterolemia: Selective Upregulation of RANTES Arterioscler Thromb Vasc Biol, January 1, 2006; 26(1): 200 - 205. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Weber Killing Two Birds With One Stone: Targeting Chemokine Receptors in Atherosclerosis and HIV Infection Arterioscler Thromb Vasc Biol, December 1, 2005; 25(12): 2448 - 2450. [Full Text] [PDF]
|
|
|
|
|
|
E. J.A. van Wanrooij, H. Happe, A. D. Hauer, P. de Vos, T. Imanishi, H. Fujiwara, T. J.C. van Berkel, and J. Kuiper HIV Entry Inhibitor TAK-779 Attenuates Atherogenesis in Low-Density Lipoprotein Receptor-Deficient Mice Arterioscler Thromb Vasc Biol, December 1, 2005; 25(12): 2642 - 2647. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Herder, B. Haastert, S. Muller-Scholze, W. Koenig, B. Thorand, R. Holle, H.-E. Wichmann, W. A. Scherbaum, S. Martin, and H. Kolb Association of Systemic Chemokine Concentrations With Impaired Glucose Tolerance and Type 2 Diabetes: Results from the Cooperative Health Research in the Region of Augsburg Survey S4 (KORA S4) Diabetes, December 1, 2005; 54(suppl_2): S11 - S17. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. A. Ali, C. A. Bursill, Y. Hu, R. P. Choudhury, Q. Xu, D. R. Greaves, and K. M. Channon Gene Transfer of a Broad Spectrum CC-Chemokine Inhibitor Reduces Vein Graft Atherosclerosis in Apolipoprotein E-Knockout Mice Circulation, August 30, 2005; 112(9_suppl): I-235 - I-241. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. R. Veillard, S. Steffens, G. Pelli, B. Lu, B. R. Kwak, C. Gerard, I. F. Charo, and F. Mach Differential Influence of Chemokine Receptors CCR2 and CXCR3 in Development of Atherosclerosis In Vivo Circulation, August 9, 2005; 112(6): 870 - 878. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Baltus, P. von Hundelshausen, S. F. Mause, W. Buhre, R. Rossaint, and C. Weber Differential and additive effects of platelet-derived chemokines on monocyte arrest on inflamed endothelium under flow conditions J. Leukoc. Biol., August 1, 2005; 78(2): 435 - 441. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. F. Mause, P. von Hundelshausen, A. Zernecke, R. R. Koenen, and C. Weber Platelet Microparticles: A Transcellular Delivery System for RANTES Promoting Monocyte Recruitment on Endothelium Arterioscler Thromb Vasc Biol, July 1, 2005; 25(7): 1512 - 1518. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Lutgens, B. Faber, K. Schapira, C. T.A. Evelo, R. van Haaften, S. Heeneman, K. B.J.M. Cleutjens, A. P. Bijnens, L. Beckers, J. G. Porter, et al. Gene Profiling in Atherosclerosis Reveals a Key Role for Small Inducible Cytokines: Validation Using a Novel Monocyte Chemoattractant Protein Monoclonal Antibody Circulation, June 28, 2005; 111(25): 3443 - 3452. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. K. Hansson Inflammation, Atherosclerosis, and Coronary Artery Disease N. Engl. J. Med., April 21, 2005; 352(16): 1685 - 1695. [Full Text] [PDF]
|
|
|
|
|
|
C. Weber Platelets and Chemokines in Atherosclerosis: Partners in Crime Circ. Res., April 1, 2005; 96(6): 612 - 616. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S.-P. Gravel and M. J. Servant Roles of an I{kappa}B Kinase-related Pathway in Human Cytomegalovirus-infected Vascular Smooth Muscle Cells: A MOLECULAR LINK IN PATHOGEN-INDUCED PROATHEROSCLEROTIC CONDITIONS J. Biol. Chem., March 4, 2005; 280(9): 7477 - 7486. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. R. Veillard, S. Steffens, F. Burger, G. Pelli, and F. Mach Differential Expression Patterns of Proinflammatory and Antiinflammatory Mediators During Atherogenesis in Mice Arterioscler Thromb Vasc Biol, December 1, 2004; 24(12): 2339 - 2344. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Collin-Osdoby Regulation of Vascular Calcification by Osteoclast Regulatory Factors RANKL and Osteoprotegerin Circ. Res., November 26, 2004; 95(11): 1046 - 1057. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Weber, A. Schober, and A. Zernecke Chemokines: Key Regulators of Mononuclear Cell Recruitment in Atherosclerotic Vascular Disease Arterioscler Thromb Vasc Biol, November 1, 2004; 24(11): 1997 - 2008. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. F. Charo and M. B. Taubman Chemokines in the Pathogenesis of Vascular Disease Circ. Res., October 29, 2004; 95(9): 858 - 866. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. A. Bursill, R. P. Choudhury, Z. Ali, D. R. Greaves, and K. M. Channon Broad-Spectrum CC-Chemokine Blockade by Gene Transfer Inhibits Macrophage Recruitment and Atherosclerotic Plaque Formation in Apolipoprotein E-Knockout Mice Circulation, October 19, 2004; 110(16): 2460 - 2466. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Kathiresan, C. Newton-Cheh, and R. E. Gerszten On the interpretation of genetic association studies Eur. Heart J., August 2, 2004; 25(16): 1378 - 1381. [Full Text] [PDF]
|
|
|
|
|
|
E. Simeoni, B. R. Winkelmann, M. M. Hoffmann, S. Fleury, J. Ruiz, L. Kappenberger, W. Marz, and G. Vassalli Association of RANTES G-403A gene polymorphism with increased risk of coronary arteriosclerosis Eur. Heart J., August 2, 2004; 25(16): 1438 - 1446. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||