逆转录病毒载体从人包装细胞系共转移人内源性逆转录病毒（HERVs）的可能性。

The potential of retroviral vectors to cotransfer human endogenous retroviruses (HERVs) from human packaging cell lines.

作者信息

Zeilfelder Udo, Frank Oliver, Sparacio Sandra, Schön Ulrike, Bosch Valerie, Seifarth Wolfgang, Leib-Mösch Christine

机构信息

Medical Clinic III, Faculty of Clinical Medicine Mannheim, University of Heidelberg, 68305 Mannheim, Germany.

出版信息

Gene. 2007 Apr 1;390(1-2):175-9. doi: 10.1016/j.gene.2006.08.019. Epub 2006 Sep 3.

DOI:10.1016/j.gene.2006.08.019

PMID:17045761

Abstract

Using a versatile and highly sensitive retroviral microarray, we have investigated particle preparations from three different human packaging cell lines harboring retroviral vector systems based on human immunodeficiency virus (HIV) and murine leukemia virus (MLV). 293Rev/Gag/Pol(i) cells inducibly express high titers of HIV-derived particles for packaging of HIV vectors. The Phoenix-GP and the Anjou 65 cell lines constitutively express MLV vector particles. We compared the transcription profiles of human endogenous retroviruses (HERVs) in all cell lines with the HERV sequences present in the particles. In addition, the influence of the transfected vector plasmid on the copackaging of HERVs was investigated. All particle preparations showed a defined pattern of endogenous retroviral sequences that differed from the cellular HERV expression pattern. HERV transcripts were observed in the particle preparations independent of whether a vector construct was coexpressed or not. Furthermore, our results suggest that particle preparations are frequently contaminated by cellular vesicles (exosomes) containing cellular RNAs including HERV transcripts.

摘要

我们使用了一种通用且高度灵敏的逆转录病毒微阵列，研究了来自三种不同人类包装细胞系的颗粒制剂，这些细胞系含有基于人类免疫缺陷病毒（HIV）和鼠白血病病毒（MLV）的逆转录病毒载体系统。293Rev/Gag/Pol(i)细胞可诱导表达高滴度的HIV衍生颗粒，用于包装HIV载体。Phoenix-GP和Anjou 65细胞系组成性表达MLV载体颗粒。我们将所有细胞系中人类内源性逆转录病毒（HERV）的转录谱与颗粒中存在的HERV序列进行了比较。此外，还研究了转染的载体质粒对HERV共包装的影响。所有颗粒制剂都显示出一种特定的内源性逆转录病毒序列模式，该模式与细胞HERV表达模式不同。无论是否共表达载体构建体，在颗粒制剂中都观察到了HERV转录本。此外，我们的结果表明，颗粒制剂经常被含有包括HERV转录本在内的细胞RNA的细胞囊泡（外泌体）污染。