丁型肝炎病毒体外短期细胞培养系统的建立

Establishment of A transient cell culture system of hepatitis D virus D in vitro

为建立ＨＤＶ的体外细胞培养系统，采用体外细胞转染技术，用含全基因ＨＤＶｃＤＮＡ三聚体的重组质粒ｐＳＶＬＤ３转染ＨＢＶ的体外细胞培养株（２．２．１５细胞株）。结果在转染后３天的转染细胞内和培养上清中检出了１．７ＫｂＨＤＶＲＮＡ和２４０００、２７０００ＨＤＡｇ。初步结果提示：该转染细胞培养上清中有ＨＤＶ病毒的排泌，从而建立了ＨＤＶ的体外短期细胞培养系统。

In order to establish the HDV cell culture system, by using in vitro transfection technique, the in vitro cell line of HBV (2.2.15 cell line) was transfected by a recombinant plasmid containing full length trimer of HDV cDNA (pSVL D3). HDV RNA was detected by spot and Northern blot hybridization using Dig HDV cDNA probe, and HDAg was detected by immunobloting method. The results showed that three days after transfection, 1.7Kb HDV RNA and 24000, 27000 HDAg were detected in the transfected cells and in its culture medium. The primary results demonstrated that hepatitis D virus was assemblied and released in the transfected cell culture medium, so an in vitro transient cell culture system of HDV had been established.