人工感染鸭源新城疫病毒对绍鸭β-防御素和细胞因子基因表达的影响

doi:10.16431/j.cnki.1671-7236.2019.02.025

摘要/Abstract

摘要：

试验旨在探讨人工感染鸭源新城疫病毒（Newcastle disease virus，NDV）对绍鸭β-防御素（avian β-defensins，AvBDs）和细胞因子基因表达的影响。选用45只20日龄SPF绍鸭，随机分为攻毒组（27只）和对照组（18只），攻毒组采用滴鼻点眼的途径（10^8.38 EID₅₀）接种鸭源NDV强毒株（Md/CH/LGD/1/2005），对照组接种磷酸盐缓冲液。在攻毒后24和48 h，从对照组及攻毒组各随机取6只鸭屠宰，分别采集肾脏、肺脏、气管、腺胃、骨髓、脾脏、盲肠扁桃体、哈德氏腺、法氏囊9个组织，运用实时荧光定量PCR法检测组织样品中9种AvBDs和细胞因子的表达量；剩余鸭每天持续观察，每隔4 d采血1次，直至24 d，检测血清抗体水平；于攻毒后24、48、72和120 h采集咽喉及泄殖腔拭子，以确定排毒周期。结果表明，感染该NDV毒株后，鸭没有临床发病症状和死亡情况发生；感染后鸭排毒开始于攻毒后第1天，到第5天停止，在攻毒组咽喉及泄殖腔拭子中均检测到NDV。抗体水平检测结果显示，该NDV能够诱导鸭体内产生抗NDV特异抗体。实时荧光定量PCR结果发现，与对照组相比，感染后24 h，部分鸭组织中AvBD1、AvBD2、AvBD5、AvBD6、AvBD9和AvBD16表达量均显著升高（P<0.05）；感染后48 h，AvBD1和AvBD6在部分鸭组织中表达量均显著上调（P<0.05）；感染后48 h法氏囊中白细胞介素6（IL-6）和脾脏中干扰素γ（IFN-γ）的表达量明显低于感染后24 h的表达量。综上所述，该鸭源NDV毒株感染可诱导绍鸭体内在早期产生天然免疫反应，这些反应可能与病毒在机体内复制有关。

关键词: 绍鸭; 新城疫病毒(NDV); 禽β-防御素(AvBDs); 细胞因子

Abstract:

To investigate the effects of artificial infected duck Newcastle disease virus (NDV) on the expression of avian β-defensins(AvBDs) and cytokines in Chinese Shaoxing duck,forty five 20-day-old SPF Chinese Shaoxing ducks were randomly divided into attack group (27) and control group (18).The ducks in attack group were intraocularly inoculated with 10^8.38 EID₅₀ of virulent NDV strain (Md/CH/LGD/1/2005),others were mock-inoculated with phosphate-buffered saline (PBS).6 ducks in two groups were killed at 24 and 48 h,respectively.Nine tissues including spleen,lung,kidney,glandular stomach,bursa of Fabricius,cecal tonsil,trachea,harderian glands and marrow,were collected from ducks for Real-time quantitative PCR analysis to quantify the mRNA levels of AvBDs and cytokines.In addition,the remaining ducks were constantly observed every day,serum was collected,and specific antibody titres was evaluated at 4,8,12,16,20 and 24 d,respectively.Oropharyngeal and cloacal swabs were collected and specific antibody titres was evaluated at 24,48,72 and 120 h,respectively.The results showed that no obvious clinical signs were observed in the NDV infected ducks.The ducks began to detoxify at the 1st day after attack,and stopped at the 5th day,NDV was detected in the oropharyngeal and cloacal swabs of attack group.The antibody level test results showed that NDV could induce anti-NDV specific antibodies in ducks.Real-time quantitative PCR results showed that compared with control group,the expression levels of AvBD1,AvBD2,AvBD5,AvBD6,AvBD9 and AvBD16 were significantly increased in some tissues of NDV infected ducks at 24 h (P<0.05);The expression levels of AvBD1 and AvBD6 were significantly up-regulated in some tissues of NDV infected ducks at 48 h (P<0.05).The expression levels of IL-6 in bursa of Fabricius and IFN-γ in spleen at 48 h were lower than that at 24 h after infection.In summary,the duck-derived NDV strain infection could induce early natural immune responses in Chinese Shaoxing duck,and these reactions might be related to the replication of the virus in the host.

Key words: Chinese Shaoxing duck; Newcastle disease virus(NDV); avian β-defensins(AvBDs); cytokines

中图分类号:

S852.65⁺9.5

许丽文, 王秋玲, 王芳芳, 刘诚刚, 马得莹. 人工感染鸭源新城疫病毒对绍鸭β-防御素和细胞因子基因表达的影响[J]. 《中国畜牧兽医》, 2019, 46(2): 536-547.

XU Liwen, WANG Qiuling, WANG Fangfang, LIU Chenggang, MA Deying. Effects of Artificially Infected Duck Newcastle Disease Virus on the Expression of β-defensins and Cytokines in Chinese Shaoxing Ducks[J]. , 2019, 46(2): 536-547.

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