乙型肝炎病毒在丁型肝炎病毒包装中的作用机理研究

Study on the role of hepatitis B virus acting on the assembly of hepatitis D virus

用含ＨＢＶ全基因和ＨＢＶ大、中、小分子表面蛋白基因的真核细胞表达质粒（ＣＭＶ－ＨＢＶ、ＣＭＶ－ＬＳ、ＣＭＶ－ＭＳ、ＣＭＶ－Ｓ）分别与含ＨＤＶ ｃＤＮＡ三聚体的重组质粒（ｐＳＶＬＤ３）共转染ＣＨＯ细胞。转染后３天在上述４种转染细胞内及培养上清中均检出了ＨＤＶＲＮＡ和ＨＤＡｇ，表明上述４种转染细胞的培养上清中均有ＨＤＶ病毒颗粒的包装和分泌。提示ＨＤＶ病毒的包装可能仅需ＨＢＶＳ基因及其小分子表面蛋白的辅助。

Hepatitis D virus (HDV ) is a defective virus that requires the presence of a helper virus , hepadnaviruses (e. g. HBV ) , for its life cycle. HDV consists of a single-stranded circular RNA of approximately 1,7kb (HDV RNA) and an internal protein ,hepatitis delta antigen (HDAg) ,surrounded by the envelope composed of surface antigen of the hepadnaviruses (e. g. HBsAg ) . Our previous study demonstrated that HDV genome could replicate and express without the assistance of hepadnaviruses , and assembly of HDV particle needs the help of HBV. But the mechanism of various HBV gene and protein acting on HDV package is still uatclear. Using transfection-reagent system , CHO cell line was transiently cotratnsfected with a recombinant plasmid containing full longth trimer of HDV cDNA (pSVLD3 ) and a plasmid containing full longth HBV DNA or HBV genome of large or middle or small surface protein (CMV-HBV , CMV-LS ,CMV-MS ,CMV- S ) . Three days after transfection , HDV RNA and HDAg were detected in the four kinds transfected cells and their culture medium mentioned above. The results demonstrated that HDV particles might have packaged and released in the medium. We thus conclude that assembly of HDV particle only needs the assistance of HBV S gene and its small surface protein.