在CHO-K1细胞系中化学诱导的内源性逆转录病毒粒子的表征
Nicholas B Mattson1, Trent J Bosma1, Yamei Gao1
1Laboratory of Retroviruses, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Review, U.S. Food and Drug Administration, Silver Spring, MD 20993, USA.
Viruses
|November 27, 2025
概括
这项研究研究了CHO-K1细胞中潜伏的传染性内源性逆转录病毒. 化学诱导激活了复原病毒颗粒,但感染性测定证实没有复制复原病毒存在.
科学领域:
- 分子生物学分子生物学
- 病毒学 病毒学
- 细胞生物学 细胞生物学
背景情况:
- 中国仓鼠卵巢K1 (CHO-K1) 细胞构成性地产生具有逆转录酶 (RT) 活性的复原病毒样颗粒 (RVLP).
- 之前的研究报告了其他动物物种的传染性逆转录病毒,促使对CHO-K1细胞进行调查.
- 内源逆转录病毒 (ERV) 可以在宿主基因组内存在潜伏状态.
研究的目的:
- 为了调查CHO-K1细胞中潜伏的,感染性内源性逆转录病毒 (ERV) 的存在.
- 为了确定化学诱导是否可以激活CHO-K1细胞中的ERV.
- 评估各种细胞系 (包括人类细胞系) 中诱导的逆转录病毒颗粒的传染性.
主要方法:
- 使用5--2'-脱氧氨和5-阿扎丁的化学诱导试验.
- 使用产品增强型RT (PERT) 试验检测逆转录酶 (RT) 活性.
- 传输电子显微镜 (TEM) 用于反病毒粒子可视化.
- 长读和短读高通量测序 (HTS) 用于病毒序列分析.
- 通过注射各种细胞系 (人,子,动物,CHO-K1) 与无细胞超级生物进行传染性研究.
主要成果:
- 化学诱导,特别是双重治疗,增加了复原病毒样粒子 (RVLP) 的产生和RT活性.
- 传输电子显微镜证实了A型和C型逆转录病毒颗粒的激活.
- 长期阅读的HTS证实了A型和C型颗粒在双重药物处理的CHO-K1细胞中的诱导.
- 传染性研究没有证据表明,在接种的细胞系中,包括人类细胞中,复制逆转录病毒.
- 短读HTS没有检测到注射人类细胞中的逆转录病毒序列集成.
结论:
- 化学诱导成功地激活了CHO-K1细胞中的RVLP和RT活性.
- 尽管激活,但在CHO-K1细胞中没有检测到传染性或复制性内源性逆转录病毒.
- 该研究没有发现CHO-K1细胞系中潜在的,传染性RVLP的证据.
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