摘要
构建了人白细胞介素－２（ＩＬ－２）基因重组逆转录病毒表达载体ｐＺＩＰｈｕｌＬ－，以ＤＮＡ－磷酸钙共沉淀技术转染包装细胞系ＰＡ３１７，以Ｇ４１８筛选，得到分泌假病毒颗粒滴度达到１０６ＣＦＵ／ｍｌ的细胞克隆。以８为感染倍数感染２．２．１５细胞系，ＩＬ－２分泌表达水平为６Ｉ／ｍ１，可显著抑制ＨＢｓＡｇ的分泌表达，对ＨＢｅＡｇ也有一定的抑制作用。但不含ＩＬ－２基因的逆转录病毒载体ｐＺＩＰＳＶ（Ｘ）的假病毒颗粒感染２．２．１５细胞以后对乙肝病毒无显著影响。以６ＩＵ／ｍｌ的重组人ＩＬ－２也不能抑制ＨＢｓＡｇ及ＨＢｅＡｇ的表达。在细胞水平上，实现了人ＩＬ－２转基因表达及抗乙肝病毒的基因治疗。
Human
interleukin-2(IL-2)expressive retroviral vector pZIPhulL- 2was
constructed.Packaging cellline PA317 was transfected by recombinant
expressive vector andG418 resistant cell clones were identi-fied by
detecting with titre of pseudoviral particles of l x l06 CFU/ml in
their culture supematants. IL-2expression level is at6IU/ml and HBsAg
expression and secretion of2.2.15 cell line was
inhibitaedsigniflcanty after pseudoviral particles infection with 8
of multiplicity of infection(M. O. I.).Howerer,neitherpseudoviral
particles containing retroviral vector pZIPSV(X),nor 6 IU/ml of
recombinant humanIL-2 had the effects on the HBsAg expression of 2.2.
15 cell line. The results indicated that IL-2 genetransfer and
expression is a potential gene therapy route for HBV infections.
