小干扰RNA表达抑制禽传染性支气管炎病毒复制及炎症反应。

Small interfering RNA expression inhibits avian infectious bronchitis virus replication and inflammatory response.

作者信息

Yu Kun, Deng Shoulong, Wang Hai, Zhang Yi, Chen Xuehui, Wang Kejun, Hu Rui, Lian Zhengxing, Li Ning

机构信息

National Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China.

Laboratory of Animal Genetics and Breeding, College of Animal Science and Technology, China Agricultural University, Beijing, China.

出版信息

Antivir Ther. 2016;21(6):469-479. doi: 10.3851/IMP3027. Epub 2016 Feb 2.

DOI:10.3851/IMP3027

PMID:26835751

Abstract

BACKGROUND

Avian infectious bronchitis virus (IBV) is a major cause of poor weight gain and mortality among chicks.

METHODS

A lentivirus vector was used to generate transgenic chickens expressing small interfering RNA (siRNA) targeting the M protein of IBV. Offspring of generation 0 (G0) were screened to identify G1 transgenic chickens (Tg). Monocytes from G1 Tg were stimulated with IBV in vitro.

RESULTS

Monocytes producing siRNA efficiently inhibit IBV replication. Expression of inflammatory cytokines, Mx protein and nitric oxide levels were lower in early IBV infection in Tg. In vivo experiments show that siRNA expression inhibits IBV replication, significantly decreases mortality and increases weight gain. Inflammatory responses and oxidative damage were significantly decreased, yielding minimal tissue injury. The inflammatory responses indicate that the cellular immune response is most effective during the initial stage, while the humoral immune response is more significant in later stages of infection.

CONCLUSIONS

Small interfering RNA expression inhibits avian IBV replication and inflammatory response.

摘要

背景

禽传染性支气管炎病毒（IBV）是导致雏鸡体重增加缓慢和死亡的主要原因。

方法

使用慢病毒载体生成表达靶向IBV M蛋白的小干扰RNA（siRNA）的转基因鸡。对第0代（G0）后代进行筛选，以鉴定第1代转基因鸡（Tg）。用IBV体外刺激G1 Tg的单核细胞。

结果

产生siRNA的单核细胞有效抑制IBV复制。在Tg中，早期IBV感染时炎性细胞因子、Mx蛋白的表达及一氧化氮水平较低。体内实验表明，siRNA表达抑制IBV复制，显著降低死亡率并增加体重。炎性反应和氧化损伤显著降低，组织损伤最小。炎性反应表明，细胞免疫反应在初始阶段最有效，而体液免疫反应在感染后期更显著。