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鸭瘟病毒糖蛋白gE在感染鸭中的表达及免疫组化分布

Expression and immunohistochemical distribution of duck plague virus glycoprotein gE in infected ducks.

作者信息

Chang Hua, Cheng Anchun, Wang Mingshu, Xiang Jun, Xie Wei, Shen Fuxiao, Jia Renyong, Zhu Dekang, Luo Qihui, Zhou Yi, Chen Xiaoyue

机构信息

Avian Diseases Research Center, College of Veterinary Medicine of Sichuan Agricultural University, Yaan, Sichuan, 625014, China.

出版信息

Avian Dis. 2011 Mar;55(1):97-102. doi: 10.1637/9487-072810-ResNote.1.

Abstract

To determine the distribution of duck plague virus (DPV) gE protein in paraformaldehyde-fixed, paraffin-embedded tissues of experimentally DPV-infected ducks, an indirect immunoperoxidase assay was established to detect glycoprotein E (gE) protein for the first time. The rabbit anti-His-gE serum, raised against the recombinant His-gE fusion protein expressed in Escherichia coli BL21 (DE3), was prepared and purified. Western blotting and indirect immunofluorescence analysis showed that the anti-His-gE serum had a high level of reactivity and specificity and could be used as the first antibody for further experiments to study the distribution of DPV gE protein in DPV-infected tissues. A number of DPV gE proteins were distributed in the bursa of Fabricius, thymus, spleen, liver, esophagus, duodenum, jejunum, ileum, and kidney of DPV-infected ducks and a few DPV gE were distributed in the Harders glands, myocardium, cerebrum, and lung, whereas the gE was not seen in the skin, muscle, and pancreas. Moreover, DPV gE was expressed 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 gE expression and distribution in infected ducks but also for understanding the pathogenesis of DPV.

摘要

为了确定鸭瘟病毒(DPV)gE蛋白在经实验感染DPV的鸭的多聚甲醛固定、石蜡包埋组织中的分布情况,首次建立了间接免疫过氧化物酶测定法来检测糖蛋白E(gE)蛋白。制备并纯化了针对在大肠杆菌BL21(DE3)中表达的重组His-gE融合蛋白产生的兔抗His-gE血清。蛋白质免疫印迹和间接免疫荧光分析表明,抗His-gE血清具有高反应性和特异性,可作为第一抗体用于进一步实验,以研究DPV gE蛋白在DPV感染组织中的分布。在感染DPV的鸭的法氏囊、胸腺、脾脏、肝脏、食管、十二指肠、空肠、回肠和肾脏中分布有大量DPV gE蛋白,在哈德氏腺、心肌、大脑和肺中分布有少量DPV gE,而在皮肤、肌肉和胰腺中未见到gE。此外,DPV gE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大量表达。本研究不仅可能有助于描述gE在感染鸭中的表达和分布特征,还可能有助于了解DPV的发病机制。

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