新城疫病毒强毒株通过感染鸡脑微血管内皮细胞增加血脑屏障渗透性来入侵鸡脑

doi:10.1016/j.jia.2024.06.003

Journal of Integrative Agriculture ›› 2025, Vol. 24 ›› Issue (2): 712-723.DOI: 10.1016/j.jia.2024.06.003

新城疫病毒强毒株通过感染鸡脑微血管内皮细胞增加血脑屏障渗透性来入侵鸡脑

收稿日期:2024-04-19 接受日期:2024-09-24 出版日期:2025-02-20 发布日期:2025-01-22

Velogenic Newcastle disease virus invades chicken brain by infecting brain microvascular endothelial cells to increase blood-brain barrier permeability

Jie Zhao^{1, 2, 3*}, Sa Xiao^2*, Wei Yao², Xudong Chang², Xinglong Wang², Zengqi Yang², Wenbin Wang^{1, 2#}

¹Poultry Institute, Shandong Academy of Agricultural Sciences, Jinan 250100, China
²College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China
³People’s Government of Guangpu Town, Chongqing 402765, China

Received:2024-04-19 Accepted:2024-09-24 Online:2025-02-20 Published:2025-01-22
About author:#Correspondence Wenbin Wang, E-mail: wangwenbin1230@163.com * These authors contributed equally to this study.
Supported by:
This work was supported by the National Natural Science Foundation of China (32302864 and 31572533), the Shandong Provincial Natural Science Foundation, China (ZR2021QC185) and the Agricultural Science and Technology Innovation Project of Shandong Academy of Agricultural Sciences, China (CXGC2023F11).

摘要/Abstract

摘要：

新城疫病毒（NDV）的嗜神经型强毒株（VNDV）引起的神经系统病变会出现非化脓性脑炎而导致显著的中枢神经紊乱，如扭颈、头颈震颤、转圈、腿或翅膀麻痹和瘫痪等症状，其雏鸡死亡率可达90%以上。生产上多见的神经型新城疫往往是鸡群发病后的后遗症，出现神经症状的家禽很难康复，未死亡的家禽长期处于疾病状态，严重影响家禽健康，并造成严重的经济损失。而VNNDV如何入侵鸡中枢神经系统的机制认不清楚。血脑屏障(BBB)可以阻止毒素和感染进入大脑。

本研究使用NDV强毒株F48E9和弱毒株LaSota分别以脑内注射和点眼滴鼻方式进行动物攻毒试验，及以鸡原代脑微血管内皮细胞（chBMECs）为细胞模型建立体外血脑屏障模型，通过病毒感染试验、细胞免疫荧光试验、荧光素钠示踪试验、透射电镜观察等评价病毒对BBB的影响。最后，通过对chBMECs使用动力蛋白抑制剂dynasore来抑制细胞的胞吞过程，研究VNDV进入该细胞的途径。

通过体内和体外实验，我们发现VNDV通过破坏血管细胞之间的紧密连接蛋白ZO-1的连续性来增加血脑屏障的通透性。根据细胞形态学与内皮细胞标记物CD31的免疫荧光染色，成功分离chBMECs细胞，通过荧光素钠渗透性检测，在transwell中成功建立体外鸡BBB模型。将强毒株F48E9和弱毒株LaSota分别感染体外BBB模型，上下层病毒滴度和荧光素钠含量的检测结果发现，只有强毒株F48E9在下层能够检测到病毒及荧光素钠，表明NDV强毒株能够增加鸡脑BBB的渗透性穿过BBB。进一步通过细胞免疫荧光染色，发现F48E9破坏chBMECs的紧密连接蛋白ZO-1之间的连接。而弱毒株LaSota对BBB没有任何影响。此外，在感染早期，使用dynamin抑制剂可降低病毒复制和血脑屏障通透性。我们的数据表明，VNDV通过感染和破坏chBMECs的紧密连接直接侵入鸡脑，从而增加血脑屏障的通透性。VNDV可通过内吞作用感染chbmec。因此，我们的研究结果为VNDV通过血脑屏障进入大脑提供了证据，为开发预防和治疗NDV的药物奠定基础。

Abstract:

The blood-brain barrier (BBB) keeps poisons and infections out of the brain. Some viruses can pass through this barrier and replicate in the central nervous system (CNS). Velogenic Newcastle disease virus (VNDV) is a neurotropic virus that causes avian nonsuppurative encephalitis. VNDV often develops into a chronic infection that seriously affects poultry health in partially immune birds. The routes by which the virus enters the chicken brain are poorly understood. In this study, we discovered that VNDV increased BBB permeability in vivo and in vitro by breaking the tight junction protein zona occludens-1 (ZO-1) continuity of chicken brain microvascular endothelial cells (chBMECs). By investigating the susceptibility of chBMECs to NDV infection, we found that VNDV but not lentogenic NDV was detected in the basolateral compartment in transwell assays after apical infection, suggesting that efficient replication and transcellular transport of the virus across the BBB in vitro. Furthermore, viral replication and BBB permeability were reduced during the early stage of infection by using the dynamin inhibitor dynasore. Our data demonstrate that VNDV invades the chicken brain by infecting and damaging the tight junction of chBMECs directly to increase BBB permeability. VNDV could infect chBMECs via endocytosis. As a result, our findings provide compelling evidence for VNDV entrance into the brain via the BBB, paving the way for the development of medications for NDV prevention and therapy.

Key words: velogenic Newcastle disease virus , blood-brain barrier , permeability , chicken brain microvascular endothelial cells

. 新城疫病毒强毒株通过感染鸡脑微血管内皮细胞增加血脑屏障渗透性来入侵鸡脑[J]. Journal of Integrative Agriculture, 2025, 24(2): 712-723.

Jie Zhao, Sa Xiao, Wei Yao, Xudong Chang, Xinglong Wang, Zengqi Yang, Wenbin Wang. Velogenic Newcastle disease virus invades chicken brain by infecting brain microvascular endothelial cells to increase blood-brain barrier permeability[J]. Journal of Integrative Agriculture, 2025, 24(2): 712-723.

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新城疫病毒强毒株通过感染鸡脑微血管内皮细胞增加血脑屏障渗透性来入侵鸡脑

Velogenic Newcastle disease virus invades chicken brain by infecting brain microvascular endothelial cells to increase blood-brain barrier permeability

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