登革热病毒诱发急性病毒性脑炎样疾病的鼠害模型
Summary
在这里,我们提出了一个协议,创建一个免疫能力ICR(癌症研究所)的中枢神经系统感染的鼠模型,以显示神经病变的发展。通过相同的疾病评分监测急性病毒性脑病,可以显示体内登革热病毒引起的神经病变。
Abstract
登革热病毒(DENV)是一种由蚊子传播的节肢传播病毒,可引起称为登革热出血热的严重疾病,其特征是血浆泄漏、腹水、胸膜渗漏、呼吸窘迫等致命并发症,严重出血和器官损伤。少数DENV感染病例有神经表现;然而,研究没有进一步探讨DENV引起的神经发病机制。在这项研究中,我们提出了一个协议,使用免疫能力超种ICR(癌症研究所)小鼠来研究中枢神经系统(CNS)感染DENV的诱导,然后急性病毒性脑炎类似进展疾病。
Introduction
DENV是Flaviviridae家族的节肢传播病毒,含有一个正义RNA基因组,该基因组编码三种病毒结构蛋白(细胞、前膜和包络)和七种病毒非结构蛋白(NS1、NS2A、NS2B、NS3、NS4A、NS4B),和 NS5)。DENV(DENV1-4)的四种血清型每年感染约3.9亿人,造成全球负担,尽管各国政府已大力控制蚊媒和疾病控制1。目前,保护性疫苗和治疗性抗病毒药物正在研制中,需要进一步的长期验证2。在临床实践中,虽然感染中枢神经系统的登革热患者很少,但还需要进一步探讨,以了解登革热发展的多样性。需要进一步调查和验证;值得注意的是,世界卫生组织(WHO)将中枢神经系统损伤(如认知障碍、抽搐、脑病和脑炎)纳入严重登革热3、4的分类。构建DENV感染的动物模型对于探索DENV感染的神经发病机制是必不可少的。
为了产生DENV的CNS感染,几项研究已经执行了不同的DENV感染途径,包括(1)接受4 x103斑块形成单位(PFU)的C57BL/6小鼠的脑内接种,6、 (2) 接受7 x 104 PFU的体外神经适应DENV47的BALB/c小鼠的腹腔内接种,(3)接受体内神经适应DENV18 1x 105 PFU的瑞士小鼠的脑内接种,以及(4)接受1 x 106 PFU的不适应DENV29的ICR吸吮小鼠的脑内和腹腔内共接种。根据这些研究的结果,小鼠的DENV感染导致病毒复制,导致急性病毒脑炎样综合征,行为变化伴随肢体瘫痪和姿势不稳定,中枢神经系统神经毒性和炎症,一般和局部血浆泄漏通过血脑屏障(BBB),和致命性。这些研究的所有结果5,6,7,8,9表明DENV感染CNS和诱导急性病毒性脑炎的能力感染后的疾病。
根据我们目前的发现9,10,11,12,13,14,15,我们已经创造了一个DENV感染的鼠模型作为体内平台,以检查靶向剂/因子对病毒复制的疗效,以及神经毒性。在这里,我们报告用于创建免疫能力的杂交ICR小鼠的协议,用于研究CNS感染,并监测由DENV引起的不同神经病变的发展。结果表明,在DENV感染的小鼠中,脑炎样疾病以时间依赖性的方式显著进展。
Protocol
Representative Results
Discussion
在3、17日,在重症登革热患者中枢神经系统中检测到DENV感染,表明在登革热发病过程中出现急性病毒性脑炎的可能性。在这里,我们报告一个在体内的DENV感染模型,以研究中枢神经系统功能障碍在严重登革热的介入,特别是侧重于DENV引起的急性病毒性脑炎样疾病。与以前的模型相比,特别是单道感染(仅限静脉注射、仅限腹腔内或仅内脑),DENV 的极高滴度(1 x 10<s…
Divulgazioni
The authors have nothing to disclose.
Acknowledgements
这项研究得到了科技部(MOST107-2321-B-038-001)和106TMU-CIT-01-2助学金的支持, 台北,台湾。
Materials
| Roswell Park Memorial Institute 1640 medium (RPMI) | Gibco | 11875-085 | Diluting virus |
| 0.3-mL Insulin Syringe | BD Ultra-FineII | 328838 | Intracerebral and intraperitoneal injection |
| Microbalance | METTLER TOLEDO's LabX | AL104 | Weight mouse |
| Non-adapted DENV2 (strain PL046) | The Centers for Disease Control of Taiwan | – | Infect mouse |
| Institute of Cancer Research (ICR) suckling mouse | BioLASCO Taiwan Co., Ltd | – | Our murine model |
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