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鸭瘟病毒 gE 蛋白在感染鸭体内的免疫荧光分析。

Immunofluorescence analysis of duck plague virus gE protein on DPV-infected ducks.

机构信息

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

出版信息

Virol J. 2011 Jan 15;8:19. doi: 10.1186/1743-422X-8-19.

Abstract

BACKGROUND

In previous studies, the expression and localization characteristics of duck plague virus (DPV) gE protein have been described in cultured cells, but the properties of DPV gE protein have not been reported in vivo. Immunofluorescence analysis had been used for the detection of virus antigen, but there was no report on the use of this technique for the detection of DPV gE. In this study, we investigated the distribution of DPV gE protein on DPV-infected ducks using polyclonal antibody raised against the recombinant His-gE fusion protein by indirect immunofluorescence assay (IFA).

RESULTS

The recombinant gE protein was highly immunogenicity by ELISA, and the gE was used as an antigen for the preparation of polyclonal antibody, which could be used the first antibody for further experiment to study the distribution of DPV gE protein in DPV-infected tissues by indirect immunofluorescence assay. DPV gE protein were distributed in the immune organs (thymus, bursa of fabricius (BF), Harders glands, spleen), the digestive organs (liver, duodenum, jejunum, ileum), and the other parenchymatous organs (kidney, myocardium, cerebrum, and lung) of DPV-infected ducks, but the positive immunofluorescence signal was not seen in the muscle and pancreas. The lymphocytes, reticulum cells, macrophages, epithelial cells, and hepatocytes served as the principal site for the localization of DPV gE antigen. Moreover, the intensity of fluorescence increased sharply from 12 to 216 h post-infection (p.i.).

CONCLUSIONS

In this work, the immunogenicity of the recombinant gE protein was analyzed by ELISA, and we presented the distribution properties of DPV gE antigen in infected ducks for the first time, which may be useful for understanding the pathogenesis of DPV. These properties of the gE protein provided the prerequisite for further functional analysis.

摘要

背景

在之前的研究中,已描述了鸭瘟病毒(DPV)gE 蛋白在培养细胞中的表达和定位特征,但尚未在体内报告 DPV gE 蛋白的特性。免疫荧光分析已用于检测病毒抗原,但尚无关于该技术用于检测 DPV gE 的报道。在这项研究中,我们使用针对重组 His-gE 融合蛋白的多克隆抗体通过间接免疫荧光测定(IFA)来研究 DPV 感染鸭中 DPV gE 蛋白的分布。

结果

ELISA 结果表明重组 gE 蛋白具有高度的免疫原性,并且 gE 蛋白被用作制备多克隆抗体的抗原,该抗体可用于进一步的实验,通过间接免疫荧光测定研究 DPV gE 蛋白在 DPV 感染组织中的分布。DPV gE 蛋白分布在免疫器官(胸腺,法氏囊(BF),哈德氏腺,脾脏),消化器官(肝脏,十二指肠,空肠,回肠)和其他实质器官(肾脏,心肌,脑和肺)中感染 DPV 的鸭子,但在肌肉和胰腺中未观察到阳性免疫荧光信号。淋巴细胞,网状细胞,巨噬细胞,上皮细胞和肝细胞是 DPV gE 抗原定位的主要部位。此外,从感染后 12 到 216 小时(p.i.),荧光强度急剧增加。

结论

在这项工作中,通过 ELISA 分析了重组 gE 蛋白的免疫原性,我们首次提出了 DPV gE 抗原在感染鸭中的分布特性,这可能有助于了解 DPV 的发病机制。gE 蛋白的这些特性为进一步的功能分析提供了前提。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/accf/3029213/e0d86c27e938/1743-422X-8-19-1.jpg

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