MECHANISMS BY WHICH CMV MAY CONTRIBUTE TO ATHEROGENESIS AND RESTENOSIS

CMV 导致动脉粥样硬化和再狭窄的机制

• 批准号: 6109286
• 负责人: J ZHU
• 金额: --
• 依托单位: NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6109286
• 关键词: Herpesviridae disease; Herpesviridae vaccine; atherosclerosis; cell migration; cellular immunity; cytomegalovirus; gene targeting; human tissue; humoral immunity; interleukin 12; laboratory mouse; laboratory rat; phenotype; restenosis; transfection; vascular smooth muscle

Increasing evidence suggests human cytomegalovirus (HCMV) infection contributes to the development of atherosclerosis. We investigated the relationship between cytomegalovirus and atherosclerosis by inoculating mice with cytomegalovirus (CMV) or virus free media. Since mice are generally resistant to the development of atherosclerosis, we used C57BL/6J apolipoprotein-e knockout mice which have been genetically engineered to develop premature atherosclerosis. Sixty mice (f=28, m=32)thirteen to fourteen days old received an intraperitoneal injection of either murine CMV (30,000 PFU)(n=30) or virus free media (n=30). Murine CMV was cultured in embryonic mouse fibroblasts. Virus free media was prepared by filtering the virus with a 0.1 um filter. All animals were kept in a sterile environment and given a regular chow diet. Half the animals were sacrificed at 13 weeks of age and the other half at sixteen weeks of age (n=15/group) since the ideal time to assess for a difference in atherosclerosis was unknown. The natural progression begins at ten weeks with fatty plaques and continues to progress to its final fibrous plaque at twenty weeks on a regular chow diet. Atherosclerosis was assessed for each mouse by examining the aortic sinus (an area at the level of the heart valves known to be predisposed to atherosclerosis in mice). An outside histologist who was blinded stained the tissues for fat and calculated the lipid surface area per cross-section. In all groups there was a trend that CMV infected mice had more atherosclerosis and the difference was greatest in older mice. At thirteen weeks of age the difference between the mean lesion area in the CMV infected group was only 9% and at sixteen weeks of age the difference was 30% (P=0.01). Subgroup analysis revealed a significant difference (P=0.04) between the female groups [mCMV=119+/-17(um x10+/-SEM;n=sections)vs. control=98+/-14] and no significant difference (P=0.09) between the male groups (54+/-8 vs. 47+/-7). Serum samples were obtained immediately prior to sacrifice and data is currently available for the sixteen-week-old animals. CRP levels (mean mg/L+/-SEM;n=4/group) in these mCMV mice (14+/-4) were higher than the control (10+/-4) but the difference was not significant. The mCMV infected mice (n=3) had the following plasma lipid levels (mean mg/dL+/-SEM); TC=638+/-86, TTG=105+/-21, HDL=23+/-4 and the control mice (n=3) had TC=632+/-114, TTG=114+/-25, and HDL=22+/-5. The CMV group had lower Gamma-IF (mean pg/mL+/-SEM;677+/-396 vs. 1024+/-579;n=4/group) and higher IL-4(mean pg/ml+/-SEM;98+/-29 vs 76+/-29) although no difference was significant. In summary, mCMV significantly increased the development of atherosclerosis in C57BL/6J apoE -/- congenic mice and the difference was greatest in sixteen-week-old females. Cytokine analysis suggests that this difference is due to a Th-2 response.

越来越多的证据表明人类 巨细胞病毒（HCMV）感染有助于发展 动脉粥样硬化我们调查了 巨细胞病毒与动脉粥样硬化的关系 巨细胞病毒（CMV）或无病毒培养基。因为老鼠是 一般抵抗动脉粥样硬化的发展，我们使用 C57 BL/6 J载脂蛋白-e基因敲除小鼠， 基因工程导致过早的动脉粥样硬化六十 小鼠（f=28，m=32）13至14天龄接受 腹腔内注射鼠CMV（30，000 PFU）（n=30） 或无病毒培养基（n=30）。小鼠CMV培养于 小鼠胚胎成纤维细胞通过以下步骤制备无病毒培养基： 用0.1 μ m过滤器过滤病毒。所有动物均饲养在 在无菌环境中并给予常规食物。一半动物 在13周龄时处死，另一半在16周龄时处死 自评估a的理想时间以来， 动脉粥样硬化的差异尚不清楚。自然进展 开始于10周的脂肪斑块，并继续发展， 其最后的纤维斑块是在20周的常规食物饮食中。 通过检查每只小鼠的动脉粥样硬化指数来评估每只小鼠的动脉粥样硬化。 主动脉窦（已知在心脏瓣膜水平的区域， 在小鼠中易患动脉粥样硬化）。一个外来的组织学家 盲法染色组织中的脂肪并计算脂质 表面积/横截面。在所有的群体中， CMV感染小鼠动脉粥样硬化发生率更高， 在年老的老鼠身上表现最好。在13周龄时， CMV感染组的平均病变面积仅为 16周龄时差异为30%（P=0.01）。 亚组分析显示有显著差异（P=0.04） 女性组之间[mCMV=119+/-17（um x10+/-SEM;n=切片）与对照组=98+/-14]，无显著性差异 男性组之间的差异（P=0.09）（54+/-8 vs. 47+/-7）。 在处死前即刻获得血清样品， 目前可获得16周龄动物的数据。CRP 这些mCMV小鼠中的水平（平均mg/L+/-SEM;n=4/组） (14+/-4）高于对照组（10+/-4），但差异 并不显著。mCMV感染的小鼠（n=3）具有 根据血浆脂质水平（平均mg/dL+/-SEM）; TC=638+/-86， TTG=105+/-21，HDL=23+/-4，对照小鼠（n=3） TC=632+/-114，TTG=114+/-25，HDL=22+/-5。的CMV 组的γ-IF较低（平均pg/mL+/-SEM;677+/-396 vs. 1024+/-579;n=4/组）和较高的IL-4（平均值 pg/ml+/-SEM;98+/-29 vs 76+/-29），但在 显著总之，mCMV显著增加了 C57 BL/6 J apoE -/-同源基因小鼠动脉粥样硬化的发生 小鼠和差异是最大的16周大的女性。 细胞因子分析表明，这种差异是由于Th-2 反应