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研究鼠白细胞介素 18(IL 18)基因的转移、表达及其抗乙型肝炎病毒 (HBV)的作用。以逆转录聚合酶链反应 (RT PCR)技术从受植物凝血素(PHA)和脂多糖 (LPS)刺激BALB/c小鼠脾脏细胞中扩增出IL 18的cDNA ,并构建表达IL 18基因的重组逆转录病毒载体pLXSN IL 18,转染PA317细胞 ,以逆转录巢式RT PCR方法 ,验证细胞培养液上清分泌的假病毒颗粒中IL 18的表达 ,将假病毒颗粒感染 2 .2 .15细胞 ,酶联免疫吸附法(ELISA)检测其上清HBsAg和HBeAg,定量检测HBVDNA。成功克隆出 5 80bp的小鼠IL 18全基因并构建逆转录病毒载体 ,在转染PA317细胞的上清中检测出含IL 18假病毒颗粒 ,上清感染 2 .2 .15细胞后第 3、5、7、14天 ,HBsAg、HBeAg逐渐下降 ,到14天时HBsAg已变为阴性 ,对HBVDNA有明显抑制作用。IL 18能成功地在逆转录病毒载体中表达 ,并有抑制HBsAg、HBeAg表达的作用
To investigate the metastasis and expression of murine interleukin 18 (IL 18) gene and its anti-hepatitis B virus (HBV) effect. IL 18 cDNA was amplified from spleen cells of BALB / c mice stimulated with phytohemagglutinin (PHA) and lipopolysaccharide (LPS) by reverse transcriptase polymerase chain reaction (RT PCR), and the IL 18 gene The recombinant retroviral vector pLXSN IL 18 was transfected into PA317 cells and the expression of IL 18 in the pseudovirions secreted by the cell culture supernatant was verified by reverse transcriptase nested RT PCR. The pseudovirions were infected with 2.2 .15 Cells and enzyme-linked immunosorbent assay (ELISA) were used to detect HBsAg and HBeAg in the supernatant, and HBVDNA was detected quantitatively. Successfully cloned 5 80bp mouse IL 18 whole genome and construct retroviral vector, detected in the supernatant of transfected PA317 cells containing IL 18 pseudovirus, the supernatant was infected with 2.2.15 cells after 3, On the 5th, 7th and 14th days, HBsAg and HBeAg decreased gradually. By the 14th day, the HBsAg had become negative and had a significant inhibitory effect on HBVDNA. IL 18 can be successfully expressed in retroviral vectors and has the effect of inhibiting the expression of HBsAg and HBeAg