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鉴定野生大象中的非洲象多瘤病毒,并创建一个表达其病毒肿瘤抗原的载体,以转化大象原代细胞。

Identification of African Elephant Polyomavirus in wild elephants and the creation of a vector expressing its viral tumor antigens to transform elephant primary cells.

机构信息

Fox Chase Cancer Center, Program in Blood Cell Development and Function, Philadelphia, Pennsylvania, United States of America.

RESIST Cluster of Excellence, Institute of Virology at Hannover Medical School, Center for Structural Systems Biology, Hamburg, Germany.

出版信息

PLoS One. 2021 Feb 5;16(2):e0244334. doi: 10.1371/journal.pone.0244334. eCollection 2021.

Abstract

Wild elephant populations are declining rapidly due to rampant killing for ivory and body parts, range fragmentation, and human-elephant conflict. Wild and captive elephants are further impacted by viruses, including highly pathogenic elephant endotheliotropic herpesviruses. Moreover, while the rich genetic diversity of the ancient elephant lineage is disappearing, elephants, with their low incidence of cancer, have emerged as a surprising resource in human cancer research for understanding the intrinsic cellular response to DNA damage. However, studies on cellular resistance to transformation and herpesvirus reproduction have been severely limited, in part due to the lack of established elephant cell lines to enable in vitro experiments. This report describes creation of a recombinant plasmid, pAelPyV-1-Tag, derived from a wild isolate of African Elephant Polyomavirus (AelPyV-1), that can be used to create immortalized lines of elephant cells. This isolate was extracted from a trunk nodule biopsy isolated from a wild African elephant, Loxodonta africana, in Botswana. The AelPyV-1 genome contains open-reading frames encoding the canonical large (LTag) and small (STag) tumor antigens. We cloned the entire early region spanning the LTag and overlapping STag genes from this isolate into a high-copy vector to construct a recombinant plasmid, pAelPyV-1-Tag, which effectively transformed primary elephant endothelial cells. We expect that the potential of this reagent to transform elephant primary cells will, at a minimum, facilitate study of elephant-specific herpesviruses.

摘要

由于象牙和身体部位的猖獗捕杀、栖息地碎片化以及人与象冲突等原因,野生大象种群数量正在迅速减少。野生象和圈养象还受到病毒的影响,包括高致病性大象内皮细胞疱疹病毒。此外,尽管古老象系的丰富遗传多样性正在消失,但由于缺乏可用于体外实验的已建立的象细胞系,大象这种癌症发病率低的物种已成为人类癌症研究中理解细胞对 DNA 损伤固有反应的一个惊人资源。然而,关于细胞对转化的抵抗力和疱疹病毒繁殖的研究受到了严重限制,部分原因是缺乏已建立的象细胞系来进行体外实验。本报告描述了一种重组质粒 pAelPyV-1-Tag 的创建,该质粒源自野生非洲象多瘤病毒(AelPyV-1)的分离株,可用于创建象细胞的永生化系。该分离株是从博茨瓦纳的一头野生非洲象(Loxodonta africana)的树干结节活检中提取的。AelPyV-1 基因组包含编码典型大(LTag)和小(STag)肿瘤抗原的开放阅读框。我们从该分离株中克隆了整个早期区,跨越 LTag 和重叠的 STag 基因,将其插入高拷贝载体中构建重组质粒 pAelPyV-1-Tag,该质粒有效地转化了原代象内皮细胞。我们预计,该试剂转化象原代细胞的潜力至少将促进对象特异性疱疹病毒的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3c93/7864673/a84b4aa6d35b/pone.0244334.g001.jpg

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