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鸭肠炎病毒UL51蛋白在实验感染鸭中的表达与分布

Expression and distribution of the duck enteritis virus UL51 protein in experimentally infected ducks.

作者信息

Shen Chanjuan, Cheng Anchun, Wang Mingshu, Xu Chao, Jia Renyong, Chen Xiaoyue, Zhu Dekang, Luo Qihui, Cui Hengmin, Zhou Yi, Wang Yin, Xu Zhiwen, Chen Zhengli, Wang Xiaoyu

机构信息

Avian Diseases Research Center, College of Veterinary Medicine of Sichuan Agricultural University, Ya'an, Sichuan 625014, China.

出版信息

Avian Dis. 2010 Jun;54(2):939-47. doi: 10.1637/9172-112109-ResNote.1.

Abstract

To determine the expression and distribution of tegument proteins encoded by duck enteritis virus (DEV) UL51 gene in tissues of experimentally infected ducks, for the first time, an immunoperoxidase staining method to detect UL51 protein (UL51p) in paraffin-embedded tissues is reported. A rabbit anti-UL51 polyclonal serum, raised against a recombinant 6-His-UL51 fusion protein expressed in Escherichia coli, was prepared, purified, and used as primary antibodies. Fifty-eight 30-day-old DEV-free ducks were intramuscularly inoculated with the pathogenic DEV CHv strain as infection group, and two ducks were selected as preinfection group. The tissues were collected at sequential time points between 2 and 480 hr postinoculation (PI) and prepared for immunoperoxidase staining. DEV UL51p was first found in the spleen and liver at 8 hr PI; in the bursa of Fabricius and thymus at 12 hr PI; in the Harders glands, esophagus, small intestine (including the duodenum, jejunum, and ileum), and large intestine (including the caecum and rectum) at 24 hr PI; in the glandularis ventriculus at 48 hr PI; and in the pancreas, cerebrum, kidney, lung, and myocardium at 72 hr PI. Throughout the infection process, the UL51p was not seen in the muscle. Furthermore, the intensity of positive staining of DEV UL51p antigen in various tissues increased sharply from 8 to 96 hr PI, peaked during 120-144 hr PI, and then decreased steadily from 216 to 480 hr PI, suggesting that the expressional levels of DEV UL51p in systemic organs have a close correlation with the progression of duck virus enteritis (DVE) disease. A number of DEV UL51p was distributed in the bursa of Fabricius, thymus, spleen, liver, esophagus, small intestine, and large intestine of DEV-infected ducks, whereas less DEV UL51p was distributed in the Harders glands, glandularis ventriculus, cerebrum, kidney, lung, pancreas, and myocardium of DEV-infected ducks. Moreover, DEV UL51p can be expressed in the cytoplasm of various types of cells, especially most abundantly in the cytoplasm of lymphocytes, reticulum cells, macrophages, epithelial cells, and hepatocytes. The present study may be useful not only for describing the characteristics of UL51p expression and distribution in vivo but also for a greater understanding of the pathogenesis of this DVE.

摘要

为确定鸭肠炎病毒(DEV)UL51基因编码的被膜蛋白在实验感染鸭组织中的表达及分布情况,首次报道了一种免疫过氧化物酶染色法用于检测石蜡包埋组织中的UL51蛋白(UL51p)。制备、纯化了针对在大肠杆菌中表达的重组6-His-UL51融合蛋白产生的兔抗UL51多克隆血清,并将其用作一抗。58只30日龄无DEV鸭肌肉注射致病性DEV CHv株作为感染组,选取2只鸭作为感染前组。在接种后(PI)2至480小时的连续时间点采集组织,制备用于免疫过氧化物酶染色的样本。DEV UL51p在接种后8小时首次在脾脏和肝脏中被发现;在接种后12小时在法氏囊和胸腺中被发现;在接种后24小时在哈德氏腺、食管、小肠(包括十二指肠、空肠和回肠)和大肠(包括盲肠和直肠)中被发现;在接种后48小时在腺胃中被发现;在接种后72小时在胰腺、大脑、肾脏、肺和心肌中被发现。在整个感染过程中,在肌肉中未发现UL51p。此外,DEV UL51p抗原在各种组织中的阳性染色强度在接种后8至96小时急剧增加,在接种后120 - 144小时达到峰值,然后在接种后216至480小时稳步下降,这表明DEV UL51p在全身器官中的表达水平与鸭病毒性肠炎(DVE)疾病的进展密切相关。大量DEV UL51p分布在DEV感染鸭的法氏囊、胸腺、脾脏、肝脏、食管、小肠和大肠中,而较少的DEV UL51p分布在DEV感染鸭的哈德氏腺、腺胃、大脑、肾脏、肺、胰腺和心肌中。此外,DEV UL51p可在各种类型细胞的细胞质中表达,尤其是在淋巴细胞、网状细胞、巨噬细胞、上皮细胞和肝细胞的细胞质中表达最为丰富。本研究不仅可能有助于描述UL51p在体内的表达和分布特征,还有助于更深入了解这种DVE的发病机制。

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