LDL Receptor-deficient Mice Research Articles

Interplay Between Apolipoprotein E and Scavenger Receptor Class B Type I Controls Coronary Atherosclerosis and Lifespan in the Mouse

The mouse has become a standard model for the study of atherosclerosis induced by extreme dyslipidemia in genetically modified strains such as the apolipoprotein E (apoE)–deficient or the LDL-receptor–deficient mouse. Although the process of plaque formation appears to repeat the fundamental steps of the human disease, it has been notoriously difficult to reproduce in the mouse the clinical consequences of atherosclerosis, such as myocardial infarction and stroke, that so commonly in patients signal plaque rupture or total lumen occlusion.1 The apoE-deficient mouse is a particularly interesting model because of its susceptibility to atherosclerotic lesions spanning from the aortic sinus to the abdominal aorta and involving the brachiocephalic and carotid arteries as well.2,3 It has recently been reported that the brachiocephalic plaque in the apoE-deficient mouse is prone to rupture on feeding a high-fat diet for periods as short as 8 weeks,4 but even in this setting of exaggerated hypercholesterolemia, the aortic sinus and coronary tree do not appear to be susceptible to plaque rupture. Scavenger receptor class B type I (SR-BI) is a scavenger-type receptor expressed by many tissues and accepted by consensus to represent the docking receptor for HDL in the liver. SR-BI-deficient mice have raised HDL levels and increased atherosclerosis susceptibility on fat feeding.5 ApoE and SR-BI are both expressed, among other cells, by the hepatocyte and the macrophage. ApoE is an efficient ligand for receptor-mediated lipoprotein removal by the liver, but it is also a strong acceptor of cellular cholesterol. As such, it may control the rate of foam cell formation in the vessel wall. Similarly, SR-BI channels cholesterol in and out of the cell, and its abundant expression by the macrophage may regulate cholesterol homeostasis in arterial foam cells. Although apoE and SR-BI play crucial roles in lipoprotein metabolism and have been …

SLE, atherosclerosis and cardiovascular disease

Atherosclerosis is an inflammatory disease and the major cause of cardiovascular disease (CVD) in general. Atherosclerotic plaques are characterized by the presence of activated immune competent cells, but antigens and underlying mechanisms causing this immune activation are not well defined. During recent years and with improved treatment of acute disease manifestations, it has become clear that the risk of CVD is very high in a prototypic autoimmune disease, systemic lupus erythematosus (SLE). SLE-related CVD and atherosclerosis are important clinical problems but may in addition also shed light on how immune reactions are related to premature atherosclerosis and atherothrombosis. A combination of traditional and nontraditional risk factors, including dyslipidaemia (and to a varying degree hypertension, diabetes and smoking), inflammation, antiphospholipid antibodies (aPL) and lipid oxidation are related to CVD in SLE. Premature atherosclerosis in some form leading to atherothrombosis is likely to be a major underlying mechanism, though distinctive features if any, of SLE-related atherosclerosis when compared with 'normal' atherosclerosis are not clear. One interesting possibility is that factors such as inflammation or aPL make atherosclerotic lesions in autoimmune disease more prone to rupture than in 'normal' atherosclerosis. Whether premature atherosclerosis is a general feature of SLE or only affects a subgroup of patients remains to be demonstrated. Treatment of SLE patients should also include a close monitoring of traditional risk factors for CVD. In addition, attention should also be paid to nontraditional risk factors such as inflammation and SLE-related factors such as aPL. Hopefully novel therapeutic principles will be developed that target the causes of the inflammation and immune reactions present in atherosclerotic lesions.

Expression of Human Myeloperoxidase by Macrophages Promotes Atherosclerosis in Mice

Myeloperoxidase (MPO) colocalizes with macrophages in the human artery wall, and its characteristic oxidation products have been detected in atherosclerotic lesions. Thus, oxidants produced by the enzyme might promote atherosclerosis. However, macrophages in mouse atherosclerotic tissue do not express MPO. Therefore, mice are an inappropriate model for testing the role of MPO in vascular disease. To overcome this problem, we generated and studied transgenic (Tg) mice that contained the human MPO gene. We produced human MPO-Tg mice with use of a Visna virus promoter. To confine MPO expression to macrophages, we lethally irradiated LDL receptor-deficient mice and repopulated their bone marrow with cells from wild-type mice or MPO-Tg mice. Despite having similarly high levels of cholesterol after maintenance on a high-fat, high-cholesterol diet, the MPO-Tg animals developed a 2-fold greater atherosclerotic area in the aorta than did mice transplanted with wild-type bone marrow (P=0.00003). Our observations indicate that expression of human MPO in macrophages promotes atherosclerosis in hypercholesterolemic mice, raising the possibility that the enzyme might be a potential therapeutic target for preventing cardiovascular disease in humans.

Mulberry (Morus alba L.) Leaves and Their Major Flavonol Quercetin 3-(6-Malonylglucoside) Attenuate Atherosclerotic Lesion Development in LDL Receptor-Deficient Mice

The effects of dietary consumption of mulberry (Morus alba L.) leaves and their major flavonol glycoside, quercetin 3-(6-malonylglucoside) (Q3MG), on the development of atherosclerotic lesions, in relation to the susceptibility of plasma LDL to oxidative modification, was studied in LDL receptor-deficient (LDLR-/-) mice. Male mice aged 8 wk were randomly assigned to 4 groups (control, quercetin, Q3MG, and mulberry). The control group was fed an atherogenic-diet containing 3 g cholesterol and 15 g cocoa butter/100 g. The other experimental groups were fed the same atherogenic diet supplemented with 0.05 g quercetin/100 g for the quercetin group, 0.05 g Q3MG/100 g for the Q3MG group, and 3 g dried mulberry-leaf powder/100 g for the mulberry group. The mice were fed their respective diets for 8 wk. The susceptibility of LDL to oxidative modification was significantly decreased in the Q3MG- and mulberry-treated mice, as evidenced by the 44.3 and 42.2% prolongation of the lag phase for conjugated diene formation compared with that of the control mice. The atherosclerotic lesion area in both the Q3MG- and mulberry-treated mice was significantly reduced by 52% compared with that of the controls. However, in the quercetin group, no protective effects were observed against LDL oxidation or atherosclerotic lesion formation. In conclusion, mulberry leaves attenuated the atherosclerotic lesion development in LDLR-/- mice through enhancement of LDL resistance to oxidative modification, and these antioxidative and antiatherogenic protective effects were attributed mainly to Q3MG, the quantitatively major flavonol glycoside in mulberry leaves.

W09-P-016 Plant sterols and stanols retard lesion progression and induce lesion regression in heterozygous LDL receptor deficient mice as well as atorvastatin

W09-P-016 Plant sterols and stanols retard lesion progression and induce lesion regression in heterozygous LDL receptor deficient mice as well as atorvastatin

Long term Rho-kinase inhibition ameliorates endothelial dysfunction in LDL-Receptor deficient mice

Chronic inhibition of Rho-kinase has been recently implicated in retardation of atherogenesis induced by high-fat diet in low-density lipoprotein receptor deficient (LDLR −/−) mice. However, it remains to be examined whether long-term Rho-kinase inhibition will reduce vascular dysfunction in this model. LDLR −/− mice on a high-fat diet were treated either with saline (LDLR −/−) or with the Rho-kinase inhibitor Fasudil (HA1077, 5-Isoquinolinesulfonyl homopiperazine, 100 mg/kg/day by gavage, LDLR −/− + Fasudil) for 10 weeks. Fasudil-treatment normalized endothelial function (measured by means of endothelium-dependent vasorelaxation) in LDLR −/− + Fasudil, to the level of controls (C57BL/6J). No tolerance toward Rho-kinase inhibition has been detected in Fasudil-treated animals. We conclude that long-term Rho-kinase inhibition normalizes endothelial function without development of tolerance.

Changes in intestinal and liver global gene expression in response to a phytosterol-enriched diet

Background: Dietary phytosterols are a recommended therapeutic option for decreasing plasma cholesterol. The increased activity of ATP-binding cassette (ABC) transporters ABCA1, ABCG5 and ABCG8, or, alternatively, a decrease in Niemann–Pick C1 Like 1 (NPC1L1) could mediate the reduction in intestinal cholesterol absorption caused by phytosterols. Other biological properties such as a direct immune modulatory activity have recently been ascribed to these plant compounds. Methods: To gain insight into the molecular effects of phytosterols, global genome-wide gene profiling and real-time RT-PCR studies were conducted in small intestines and livers of phytosterol-treated apolipoprotein E-deficient (apoE −/−) mice. Re-testing of the main results was performed in C57BL/6J and LDL receptor-deficient (LDLR −/−) mice. Results: Intestinal cholesterol absorption was decreased in all mouse models but plasma cholesterol was only decreased in apoE −/− and LDLR −/− mice. ABCA1, ABCG5, ABCG8 and NPC1L1 mRNA levels were slightly reduced in the intestine of phytosterol-treated apoE −/− and LDLR −/− mice, but increased in C57BL/6J-treated mice. Phytosterols changed genes involved in immune regulation in apoE −/− mice. However, these changes were less extensive in LDLR −/− mice and were not found in C57BL/6J mice. Conclusions: Inhibition of intestinal cholesterol absorption by phytosterols is not mediated via transcriptional changes in ABCA1, ABCG5, ABCG8 or NPC1L1. Changes suggestive of immunomodulation are associated with the hypocholesterolemic effect of phytosterols and with apoE deficiency.

Macrophage-specific overexpression of group IIa sPLA2 increases atherosclerosis and enhances collagen deposition

Atherosclerosis is a chronic inflammatory disease of the vessel wall characterized by the accumulation of lipid-laden macrophages and fibrotic material. The initiation of the disease is accompanied by the accumulation of modified lipoproteins in the vessel wall. Group IIa secretory phospholipase A2 (sPLA2 IIa) is a key candidate player in the enzymatic modification of low density lipoproteins. To study the role of sPLA2 IIa in macrophages during atherogenesis, transgenic mice were generated using the human sPLA2 IIa gene and the CD11b promoter. Bone marrow transplantation to LDL receptor-deficient mice was performed to study sPLA2 IIa in atherosclerosis. After 10 weeks of high-fat diet, mice overexpressing sPLA2 IIa in macrophages showed 2.3-fold larger lesions compared with control mice. Pathological examination revealed that sPLA2 IIa-expressing mice had increased collagen in their lesions, independent of lesion size. However, smooth muscle cells or fibroblasts in the lesions were not affected. Other parameters studied, including T-cells and cell turnover, were not significantly affected by overexpression of sPLA2 IIa in macrophages. These data clearly show that macrophage sPLA2 IIa is a proatherogenic factor and suggest that the enzyme regulates collagen production in the plaque and thus fibrotic cap development.

Lesion Macrophages Are a Key Target for the Antiatherogenic Effects of LXR Agonists

The Liver X Receptors (LXRα and LXRβ) are members of the nuclear hormone receptor superfamily.1 These sterol-responsive transcription factors regulate the expression of a number of genes involved in intestinal cholesterol absorption, conversion of cholesterol to bile acids, and reverse cholesterol transport. Activation of LXRs blocks cholesterol absorption and induces cholesterol efflux from lipid-loaded cells such as macrophages. Because increased cholesterol efflux is predicted to limit the transformation of macrophages into atherosclerotic foam cells, LXR activity is predicted to be antiatherogenic. This hypothesis is supported by several studies using mice genetically deficient in LXR expression. Previous work from Tangirala and colleagues established that selective elimination of macrophage LXRα and LXRβ expression in the bone marrow compartment results in accelerated disease in both the spontaneous (ApoE−/−) and Western-diet inducible (LDLR−/−) models of atherosclerosis.2 Furthermore, LXRαβ double knockout mice fed a normal chow diet exhibit increased cholesterol accumulation in the macrophages of their arterial walls.2,3 In addition to their effects on cholesterol metabolism, activation of the LXRs by synthetic agonists has an inhibitory effect on inflammatory gene expression in macrophages, pointing to a second potential antiatherogenic mechanism of these receptors.4 Finally, recent studies suggest that the ability of microbial infections to modify the development of atherosclerosis may be accomplished in part through interference with the LXR signaling pathway and macrophage cholesterol metabolism.5 See page 135 Collectively, these studies point to LXRs as potential therapeutic targets for the treatment or prevention of atherosclerosis. However, it …

Effects of diabetes and CETP expression on diet‐induced atherosclerosis in LDL receptor‐deficient mice

The role of CETP expression and diabetes in atherogenesis was investigated in mice with heterozygous disruption of the LDL receptor gene (LDLR1). LDLR1 mice with and without CETP expression were treated with streptozotocin (STZ) and maintained on a standard diet for one month before switching to an atherogenic diet for an additional month. STZ-sensitive mice had approximately 2.5-fold higher glycemia and 7.5- to 8.0-fold higher cholesterolemia. Factorial analysis of variance showed no significant effect of diabetes, CETP or diabetes-CETP interaction on the size of the atherosclerotic lesions. CETP expression in non-diabetic mice resulted in a 50% reduction in the area of the atherosclerotic lesions. Multiple regression analysis showed a positive and independent atherogenic effect of triglyceridemia in LDLR1 mice and of cholesterolemia in diabetic mice. Logistic analysis showed that elevated plasma cholesterol level significantly increased the risk of developing large lesion size (>75th percentile). In conclusion, CETP expression did not alter the lesion formation in response to diabetes, although it may be protective in the euglycemic state; the triglyceride level was an independent risk factor for LDL receptor-deficient mice but not for CETP-expressing mice; and elevated plasma cholesterol levels increased the risk of developing large atherosclerotic lesions, independently of CETP and diabetes.

Hypercholesterolemia Stimulates Angiotensin Peptide Synthesis and Contributes to Atherosclerosis Through the AT 1A Receptor

Hypercholesterolemia-induced atherosclerosis is attenuated by either pharmacological antagonism of AT1 receptors or AT1A receptor deficiency. However, the mechanism underlying the pronounced responses to angiotensin II (Ang II) antagonism has not been determined. We hypothesized that hypercholesterolemia stimulates the production of angiotensin peptides to provide a rationale for the profound effect of AT1A receptor deficiency on atherogenesis. Atherosclerotic lesions were analyzed in LDL receptor-deficient mice. Immunocytochemical analysis demonstrated that atherosclerotic lesions contained all the components of the conventional pathway for Ang II synthesis. AT1A receptor deficiency caused a marked decrease in atherosclerotic lesion size in both the aortic root and arch of male and female mice, without a discernible effect on composition. AT1A receptor deficiency-induced reductions in atherosclerosis were independent of systolic blood pressure and measurements of oxidation and chemoattractants. Aortic AT2 receptor mRNA expression was not altered in AT1A receptor-deficient mice, and AT2 receptor deficiency had no effect on lesion area or cellular composition. Hypercholesterolemia greatly augmented the systemic renin-angiotensin system, as demonstrated by large increases in plasma concentrations of angiotensinogen and angiotensin peptides (Ang II, III, IV, and 4-8). These increases were ablated in hypercholesterolemic AT1A receptor-deficient mice. AT1A receptor deficiency had a striking effect in reducing hypercholesterolemia-induced atherosclerosis in LDL receptor-negative mice. Hypercholesterolemia was associated with increased systemic angiotensinogen and angiotensin peptides, which were reduced in AT1A receptor-deficient mice. These results demonstrate that hypercholesterolemia-induced stimulation of angiotensin peptide production provides a basis for the marked effect of AT1A receptor deficiency in reducing atherosclerosis.

Hyperlipidemia in Concert With Hyperglycemia Stimulates the Proliferation of Macrophages in Atherosclerotic Lesions

Hyperglycemia and hyperlipidemia are important risk factors for diabetes-accelerated atherosclerosis. Macrophage proliferation has been implicated in the progression of atherosclerosis. We therefore investigated the effects of hyperglycemia and hyperlipidemia on macrophage proliferation in murine atherosclerotic lesions and isolated primary macrophages. Hyperglycemic LDL receptor-deficient mice that were fed a cholesterol-free diet for 12 weeks did not have elevated cholesterol levels compared with nondiabetic mice, and there was no evidence of increased macrophage proliferation in atherosclerotic lesions. Moreover, elevated glucose levels did not increase proliferation of isolated mouse peritoneal macrophages. In contrast, hyperglycemic LDL receptor-deficient mice that were fed a cholesterol-rich diet showed increased cholesterol levels concomitant with macrophage proliferation in atherosclerotic lesions. Glucose promoted lipid and protein oxidation of LDL in vitro. Glucose-oxidized LDL resulted in phosphorylation of extracellular signal-regulated kinase and protein kinase B/Akt and stimulated proliferation of isolated macrophages. The mitogenic effect of glucose-oxidized LDL was mediated by CD36 and by extracellular signal-regulated kinase activation induced by protein kinase C-dependent and phosphatidylinositol 3-kinase-dependent pathways. Thus, hyperglycemia is not sufficient to stimulate macrophage proliferation in lesions of atherosclerosis or in isolated macrophages. A combination of hyperglycemia and hyperlipidemia, however, stimulates macrophage proliferation by a pathway that may involve the glucose-dependent oxidation of LDL.

Are Anti–Oxidized Low-Density Lipoprotein Antibodies Pathogenic or Protective?

Despite a recent decline, atherosclerosis remains the most common cause of death in the Western world. The disease course of atherosclerosis is characterized by its chronicity, with progression in its initial stages being particularly insidious. Chronic inflammation is the pathological hallmark of atherosclerosis,1–4 and inflammatory processes are instrumental at all stages of this disease. Even before the development of detectable intimal lesions, the expression pattern of the endothelium has been shown to be inflammatory in nature, conforming to the response-to-injury hypothesis first postulated by the late Russell Ross.5 Thus, in lesion-prone sites of the arterial tree, the endothelial expression of adhesion molecules is upregulated, reflecting endothelial dysfunction secondary to unfavorable blood rheology6 and/or hypercholesterolemia.7–9 Atherosclerosis is known today to be associated with the burden of infection as well.10 The presence of infectious/inflammatory pathogeneses raises the autoimmune aspect of the condition.11–14 Anti–oxidized LDL (oxLDL) antibodies are exceptional among the autoantibodies prevalent in atherosclerosis: on the one hand, a correlation was found between the existence and titers of anti-oxLDL antibodies and the extent of atherosclerosis and cardiovascular disease (CVD) (Table 1). On the other hand, experimental data indicate that anti-oxLDL antibodies may be protective. View this table: TABLE 1. Correlation Between Titers of Anti-oxLDL Abs and Atherosclerosis and CVD oxLDL frequently presents in the sera of patients with autoimmune conditions, acute coronary syndrome, or stable coronary artery disease (CAD).15–18 OxLDL has been associated with both subclinical atherosclerosis and inflammatory variables.19 A large amount of oxLDL accumulates in atherosclerotic plaques,20–22 and the serum concentrations of circulating oxLDL may correlate with the severity of CAD23 and acute coronary syndrome.24 OxLDL seems to be an immunogenic molecule that stimulates the induction of anti-oxLDL antibodies. Thus, it is not surprising that an association was found between the presence …

Overexpression of 15-Lipoxygenase in the Vascular Endothelium Is Associated with Increased Thymic Apoptosis in LDL Receptor-Deficient Mice

Background: 15-Lipoxygenase (15-LO) is a nonheme iron-containing enzyme that catalyzes the peroxidation of fatty acids. Herein, we studied the effect of 15-LO overexpression in the vascular endothelium on thymocyte apoptosis by evaluating thymuses from low-density lipoprotein receptor-deficient (LDL-RD) mice and LDL-RD/15-LO mice. Thymuses were evaluated by immunhistochemistry and by TUNEL whereas in vitro studies were carried out by employing freshly isolated thymocytes from the respective mice and evaluation of apoptosis by propidium iodide and annexin V cytometry. Methods and Results: The apoptotic index in LDL-RD/15-LO mice was significantly higher than in the LDL-RD mice. In the thymic medulla the difference was smaller, although still significant. Freshly isolated thymus cells from LDL-RD/15-LO mice exhibited a higher rate of spontaneous cell death than controls. Incubation of thymus cells in the presence of the cell-permeable caspase-3 inhibitor DEVD-CMK resulted in a decrease in the frequency of apoptotic cells in LDL-RD/15-LO thymocytes, whereas no effect was evident in control thymocytes. The antioxidant N-acetylcysteine causes the increase in apoptosis in both groups. Conclusion: LDL-RD/15-LO mice exhibit increased thymocyte apoptosis both in vivo and in vitro. These findings may suggest a role for 15-LO in the natural selection of thymocytes.

IL-5 links adaptive and natural immunity in reducing atherosclerotic disease

Oxidized LDL induces changes in several facets of the immune system, although the relationships between these facets and their contributions to atherogenesis have yet to be fully elucidated. A report in this issue of the JCI provides a novel demonstration of the adaptive immune system influencing the production of natural antibodies. The results demonstrate that injection of malondialdehyde-modified LDL promotes a Th2 response that in turn increases the titers of the natural antibody T15/EO6, which recognizes the oxidized phospholipid POVPC. Atherosclerotic lesion size in LDL receptor–deficient mice is reduced as a consequence of the increase in natural antibody titers, and IL-5 is identified as the link between the adaptive and natural immune systems.

The recycling of apolipoprotein E and its amino-terminal 22 kDa fragment: evidence for multiple redundant pathways

A portion of apolipoprotein E (apoE) internalized by hepatocytes is spared degradation and is recycled. To investigate the intracellular routing of recycling apoE, primary hepatocyte cultures from LDL receptor-deficient mice and mice deficient in receptor-associated protein [a model of depressed expression of LDL receptor-related protein (LRP)] were incubated with human VLDL containing 125I-labeled human recombinant apoE3. Approximately 30% of the internalized intact apoE was recycled after 4 h. The N-terminal 22 kDa fragment of apoE was also resecreted, demonstrating that this apoE domain contains sufficient sequence to recycle. The 22 kDa fragment has reduced affinity for lipoproteins, suggesting that apoE recycling is linked to the ability of apoE to bind directly to a recycling receptor. Finally, apoE was found to recycle equally well in the presence of brefeldin A, a drug that blocks transport from the endoplasmic reticulum and leads to collapse of the Golgi stacks. Our studies demonstrate that apoE recycling occurs 1) in the absence of the LDL receptor or under conditions of markedly reduced LRP expression; 2) when apoE lacks the carboxyl-terminal domain, which allows binding to the lipoprotein; and 3) in the absence of an intact Golgi apparatus. We conclude that apoE recycling occurs through multiple redundant pathways.

Participation of macrophages in atherosclerotic lesion morphology in LDLr−/− mice

Lystbeige (beige) mice crossed with LDL receptor-deficient (LDLr-/-) mice had a distinct atherosclerotic lesion morphology that was not observed in LDLr-/- mice. This morphology is often associated with a stable plaque phenotype. We hypothesized that macrophage expression of the beige mutation accounted for this distinct morphology. Cultured bone marrow-derived macrophages from LDLr-/- and beige,LDLr-/- mice were compared for their ability to accumulate cholesterol, efflux cholesterol, migrate in response to chemotactic stimuli through Matrigel-coated membranes, and express matrix metalloproteinase 9 (MMP9). No differences in cholesterol metabolism were identified. Beige,LDLr-/- macrophage invasion in vitro appeared to be less than LDLr-/- macrophage invasion but did not achieve significance. Nevertheless, tumor necrosis factor-alpha-induced MMP9 expression, secretion, and enzymatic activity of beige,LDLr-/- macrophages were all significantly decreased compared with those of LDLr-/- macrophages (P < 0.05). For in vivo analyses of macrophage function, bone marrow transplantation (BMT) studies were performed. LDLr-/- mice and beige,LDLr-/- mice were irradiated and reconstituted with wild-type or beige bone marrow from mice expressing green fluorescent protein (GFP). Identification of GFP cells provided for direct identification of donor-derived cells within lesions. Only expression of the beige mutation in the BMT recipients altered the macrophage location and collagen content of the lesions. These results suggested that impaired macrophage function by itself did not account for the stable lesion morphology of beige,LDLr-/- double-mutant mice.

Characterization of apoM in normal and genetically modified mice

A novel human apolipoprotein [apolipoprotein M (apoM)] was recently described and demonstrated to be a lipocalin. We have now examined apoM in wild-type mice and mice with genetically altered lipoprotein metabolism. Liver and kidney showed high mRNA expression, whereas spleen, heart, brain, and testis demonstrated low expression. ApoM gene expression was initiated on embryonic day 10. Western blot analysis of plasma suggested that mouse apoM, like its human counterpart, is secreted with a retained signal peptide, but unlike human apoM it is not glycosylated. Gel filtration of plasma showed apoM to be associated with HDL-sized particles in wild-type and apoA-I-deficient mice and with HDL- and LDL-sized particles in LDL receptor-deficient mice, whereas apoM was mainly found in VLDL-sized particles in high-fat, high-cholesterol-fed apoE-deficient mice. The plasma concentration of apoM was similar in wild-type, LDL receptor-deficient, and apoE-deficient mice but was reduced to 33% in apoA-I-deficient compared with wild-type mice (P = 0.007). These data suggest that apoM mainly associates with HDL in normal mice but also with the pathologically increased lipoprotein fraction in genetically modified mice. The substantially decreased apoM levels in apoA-I-deficient mice suggest a connection between apoM and apoA-I metabolism.

Suppression of early atherosclerosis in LDL-receptor deficient mice by oral tolerance with $beta;2-glycoprotein I

Background: Atherosclerosis is considered analogous to chronic inflammatory diseases. Beta2-glycoprotein I (β2GPI) is a phospholipid binding protein shown to serve as a target for prothrombotic antiphospholipid antibodies. It has recently been demonstrated to drive an immune mediated reaction and enhance murine atherosclerosis. Oral tolerance is a method in which feeding a given antigen, downregulates the respective immune responses towards it, and attenuates concomitant organ specific disorders. Herein, we tested the hypothesis, that inhibiting cellular immunity to β2GPI would result in suppression of fatty streak formation in mice. Methods and results: LDL receptor deficient mice were fed different doses of human or bovine β2GPI or BSA and than switched to an atherogenic diet. To determine the effect of feeding on lymph node proliferative indices, separate groups of mice were fed β2GPI and then immunized with the respective antigen. Feeding either human or bovine β2GPI was effective in attenuating atherosclerosis as compared to control fed animals. Oral feeding with of β2GPI inhibited lymph node cell reactivity to β2GPI in mice immunized against the human protein. Oral tolerance was also capable of reducing reactivity to oxidized LDL in mice immunized against oxLDL. IL-4 and IL-10 production was upregulated in lymph node cells of β2GPI-tolerant mice immunized against β2GPI, upon priming with the respective protein. Conclusion: Thus, oral administration of β2GPI is an effective means of suppressing atherogenesis in mice and should further be investigated.

Suppression of early atherosclerosis in LDL-receptor deficient mice by oral tolerance with beta 2-glycoprotein I.

Atherosclerosis is considered analogous to chronic inflammatory diseases. Beta 2-glycoprotein I (beta 2GPI) is a phospholipid binding protein shown to serve as a target for prothrombotic antiphospholipid antibodies. It has recently been demonstrated to drive an immune mediated reaction and enhance murine atherosclerosis. Oral tolerance is a method in which feeding a given antigen, downregulates the respective immune responses towards it, and attenuates concomitant organ specific disorders. Herein, we tested the hypothesis, that inhibiting cellular immunity to beta 2GPI would result in suppression of fatty streak formation in mice. LDL receptor deficient mice were fed different doses of human or bovine beta 2GPI or BSA and than switched to an atherogenic diet. To determine the effect of feeding on lymph node proliferative indices, separate groups of mice were fed beta 2GPI and then immunized with the respective antigen. Feeding either human or bovine beta 2GPI was effective in attenuating atherosclerosis as compared to control fed animals. Oral feeding with of beta 2GPI inhibited lymph node cell reactivity to beta 2GPI in mice immunized against the human protein. Oral tolerance was also capable of reducing reactivity to oxidized LDL in mice immunized against oxLDL. IL-4 and IL-10 production was upregulated in lymph node cells of beta 2GPI-tolerant mice immunized against beta 2GPI, upon priming with the respective protein. Thus, oral administration of beta 2GPI is an effective means of suppressing atherogenesis in mice and should further be investigated.