DEVELOPMENTAL EXPRESSION OF COXSACKIE ADENOVIRUS RECEPTOR

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 前言：我们进行了一项初步研究，以建立肠道病毒感染和心肌炎的非人类灵长类动物模型。方法：将从致死性心肌炎患者的临床标本中分离到的柯萨奇病毒B3(CVB-3)或柯萨奇病毒B3/肠道病毒86株(CVB-MCH)分别经口服和静脉途径接种食蟹猴幼猴(n=1)。在基线、第3、7、14和28天进行临床和超声心动图评估。检测CVB-3中和抗体效价。采用逆转录-聚合酶链式反应(RT-PCR)及血浆和心脏组织培养检测病毒RNA。第14天或28天处死动物，对心肌进行病毒病理学检查。结果：两种CVB-3毒株均通过两种途径获得感染。6/9出现病毒血症，3天后消退，全部(9/9)产生中和抗体。感染CVB-3107个感染单位(Iu)的两只动物分别于第11天和第14天死亡，随后接种106Iu的CVB-3动物一直表现良好，直到第14天(n=3)和第28天(n=1)被处死。感染106IU CVB-MCH的3只也表现良好，直到第28天处死。射血分数维持正常，但4只动物有轻微的室壁运动异常。血清肌钙蛋白保持正常。5/5的受试者心肌内有病毒存在。所有动物均有多个部位的心肌损伤，包括淋巴细胞性或嗜酸性心肌炎(n=6)，局灶性炎症(n=3)，肌细胞溶解(n=2)，缺血灶(n=4)，收缩带坏死(n=2)和肥大(n=3)。第14天有4/5的动物出现活动性心肌炎，第28天有2/4的动物出现活动性心肌炎。结论：我们能够一致地在食蟹猴体内建立CVB-3感染，导致临床明显或亚临床疾病，包括持续28天的心肌炎或心脏损伤。尽管对清除病毒血症有足够的免疫反应，病毒仍在心肌中持续存在。28d时出现的心肌细胞肥大和损伤可能为免疫或病毒诱导的心肌病的调控实验提供了基础。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Introduction: We performed a pilot study to establish a non-human primate model of enteroviral infection and myocarditis. Methods: Juvenile cynomolgus monkeys were inoculated via oral (n=1) or intravenous (n=8) routes with a cloned Coxsackievirus B3 (CVB-3) or a CVB-3 /Enterovirus 86 strain (CVB-MCH) isolated from a clinical specimen from a patient with fatal myocarditis. Clinical and echocardiographic assessment was performed at baseline and at days 3, 7, 14, and 28. CVB-3 neutralizing antibody titers were measured. Viral RNA was detected by RT-PCR and culture of plasma and cardiac tissue. Animals were sacrificed at 14 or 28 days and myocardium examined for viral pathology. Results: Infection was achieved by both routes with both CVB-3 strains. Viremia was detected in 6/9, resolving after day 3. All (9/9) generated neutralizing antibody. Two animals infected with 107 infectious units (IU) of CVB-3 became moribund and were sacrificed at day 11 and day 14. Subsequent animals received 106 IU of CVB-3 and appeared well until sacrificed at day 14 (n=3) or day 28 (n=1). Three infected with 106 IU CVB-MCH also appeared well until sacrificed at day 28. Normal ejection fraction was maintained, but four animals had minor wall motion abnormalities. Serum troponin remained normal. Virus was present in myocardium in 5/5 tested. Multiple areas of focal cardiac injury were present in all animals and included lymphocytic or eosinophilic myocarditis (n=6), focal inflammation (n= 3), myocytolysis (n=2), ischemic foci (n=4), contraction band necrosis (n=2), and hypertrophy (n=3). Active myocarditis was present in 4/5 animals at day 14 and 2/4 at day 28. Conclusions: We were able to consistently establish infection with CVB-3 in cynomolgus monkeys resulting in clinically apparent or sub clinical disease, including myocarditis or cardiac injury persisting at 28 days. Despite adequate immune responses to clear viremia, virus persisted in the myocardium. The presence of myocyte hypertrophy and injury at 28 days in this model may provide a basis for experiments aimed at modulating immune or virus induced cardiomyopathy.