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以PCR技术扩增得到乙肝病毒(ayw型)表面抗原(HBsAg)的编码基因,构建S基因真核表达载体pcDNA3.1(+)-S(以下简称S)。以PCR方法扩增得到小鼠白细胞介素-12的全长cDNA序列,构建真核表达载体pcDNA3-IL-12(以下简称IL-12)。以S和S+IL-12经肌肉注射免疫BALB/C小鼠,以空载体pcDNA3.1(+)为阴性对照,血源HBsAg蛋白为阳性对照,用ELISA方法检测不同时间免疫小鼠血清中的抗-HBs。结果表明:S单独和S+IL-12联合免疫2周后,血清中抗-HBs阳转。同时用IL-12免疫,使抗体达峰时间缩短,抗体持续时间缩短,滴度降低。因此,S基因免疫可刺激产生特异性体液免疫;联合IL-12基因免疫可抑制S基因免疫诱生的体液免疫应答。
The gene encoding hepatitis B virus surface antigen (HBsAg) was amplified by PCR and the eukaryotic expression vector pcDNA3.1 (+) - S (hereinafter referred to as S) was constructed. The full-length cDNA sequence of mouse interleukin-12 was amplified by PCR and the eukaryotic expression vector pcDNA3-IL-12 (hereinafter referred to as IL-12) was constructed. BALB / C mice were immunized with S and S + IL-12 by intramuscular injection. Negative control of empty vector pcDNA3.1 (+) and HBsAg protein of blood were used as positive control. Serum Anti-HBs. The results showed that anti-HBs positive in serum after S combined with S + IL-12 for 2 weeks. At the same time with IL-12 immunization, the antibody peak time shortened antibody duration shortened titers. Therefore, S gene immunization can stimulate the production of specific humoral immunity; combined with IL-12 gene immunization can inhibit S gene immunity-induced humoral immune response.