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深度测序揭示了两种感染生态系统中禽白血病病毒主要变体之间的显著多样性和进化动态。

A deep sequencing reveals significant diversity among dominant variants and evolutionary dynamics of avian leukosis viruses in two infectious ecosystems.

作者信息

Meng Fanfeng, Dong Xuan, Hu Tao, Chang Shuang, Fan Jianhua, Zhao Peng, Cui Zhizhong

机构信息

College of Veterinary Medicine, Shandong Agricultural University, Taian, 271018, China.

Institute of Pathogen Biology, Taishan Medical College, Taian, Shandong, China.

出版信息

BMC Vet Res. 2016 Dec 19;12(1):287. doi: 10.1186/s12917-016-0902-6.

Abstract

BACKGROUND

As a typical retrovirus, the evolution of Avian leukosis virus subgroup J (ALV-J) in different infectious ecosystems is not characterized, what we know is there are a cloud of diverse variants, namely quasispecies with considerable genetic diversity. This study is to explore the selection of infectious ecosystems on dominant variants and their evolutionary dynamics of ALV-J between DF1 cells and specific-pathogen-free (SPF) chickens. High-throughput sequencing platforms provide an approach for detecting quasispecies diversity more fully.

RESULTS

An average of about 20,000 valid reads were obtained from two variable regions of gp85 gene and LTR-U3 region from each sample in different infectious ecosystems. The top 10 dominant variants among ALV-J from chicken plasmas, DF1 cells and liver tumor were completely different from each other. Also there was a difference of shannon entropy and global selection pressure values (ω) in different infectious ecosystems. In the plasmas of two chickens, a large portion of quasispecies contained a 3-peptides "LSD" repeat insertion that was only less than 0.01% in DF1 cell culture supernatants. In parallel studies, the LTR-U3 region of ALV-J from the chicken plasmas demonstrated more variants with mutations in their transcription regulatory elements than those from DF1 cells.

CONCLUSIONS

Our data taken together suggest that the molecular epidemiology based on isolated ALV-J in cell culture may not represent the true evolution of virus in chicken flocks in the field. The biological significance of the "LSD" insert and mutations in LTR-U3 needs to be further studied.

摘要

背景

作为一种典型的逆转录病毒,禽白血病病毒J亚群(ALV-J)在不同感染生态系统中的进化特征尚不明确,我们只知道存在大量多样的变体,即具有相当遗传多样性的准种。本研究旨在探索感染生态系统对DF1细胞和无特定病原体(SPF)鸡之间ALV-J优势变体的选择及其进化动态。高通量测序平台提供了一种更全面检测准种多样性的方法。

结果

从不同感染生态系统中每个样本的gp85基因和LTR-U3区域的两个可变区平均获得约20,000条有效 reads。来自鸡血浆、DF1细胞和肝肿瘤的ALV-J中排名前十的优势变体彼此完全不同。不同感染生态系统中的香农熵和全局选择压力值(ω)也存在差异。在两只鸡的血浆中,大部分准种含有一个3肽“LSD”重复插入,而在DF1细胞培养上清液中该插入仅占不到0.01%。在平行研究中,来自鸡血浆的ALV-J的LTR-U3区域在其转录调控元件中显示出比来自DF1细胞的更多具有突变的变体。

结论

我们的数据综合表明,基于细胞培养中分离的ALV-J的分子流行病学可能无法代表该病毒在田间鸡群中的真实进化情况。“LSD”插入和LTR-U3突变的生物学意义需要进一步研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/036b/5168851/7439415313d1/12917_2016_902_Fig1_HTML.jpg

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