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目的:观察核酸疫苗预敏,乙型肝炎病毒HBsAg蛋白疫苗增强的免疫对Balb/c小鼠免疫应答的影响。方法:以Transfection TM脂质体将乙型肝炎病毒表面抗原中蛋白(MHBs)核酸疫苗pSW3891/MHBs/ad(r简称为adr),及空载体pSW3891(简称vector)体外转染293T细胞。免疫印迹法(Westernblot)检测adr,vector的体外表达;动物体内研究选用Balb/c小鼠共18只,每组6只,编号后随机分为3组,即空载体质粒组(vector+vector组)、adr核酸疫苗+HBsAg蛋白疫苗(adr+protein组)、HBsAg蛋白疫苗+HBsAg蛋白疫苗(Protein+Protein组);于第0周肌肉注射法分别以vector、adr及HBsAg蛋白疫苗免疫小鼠,于第4周肌内注射法分别以vector、HBsAg蛋白疫苗、及HBsAg蛋白疫苗免疫小鼠。采用酶联免疫吸附试验(ELISA)检测小鼠血清HBsAg特异性抗体、酶联免疫斑点(ELISPOT)法检测小鼠脾细胞HBsAg多肽特异性IFN-γ分泌细胞。结果:adr体外转染293T细胞后,能够表达乙型肝炎病毒表面抗原中蛋白(MHBs);体内研究结果显示:除vector+vector组外,adr+protein组、Protein+Protein组小鼠均能检出血清抗-HBs,Protein+Protein组抗-HBs终点滴度比adr+protein组高,但无统计学意义;三组中vector+vector组没有检测到特异性INF-γ分泌的脾细胞,而adr+protein组、Protein+Protein组小鼠均能检出,且adr+protein组特异性细胞数量显著高于protein+protein组(P<0.001),具有统计学意义。结论:乙型肝炎病毒表面抗原中蛋白(MHBs)核酸疫苗预敏,能明显增强Balb/c小鼠对乙型肝炎HBsAg蛋白疫苗细胞免疫应答水平。
OBJECTIVE: To observe the effect of nucleic acid vaccine-pre-sensitized and boosted immunity of hepatitis B virus HBsAg vaccine on immune response in Balb / c mice. Methods: The recombinant plasmid pSW3891 / MHBs / ad (abbreviated as adr) and empty vector pSW3891 (vector) were transfected into 293T cells with Transfection TM liposome. The expression of adr and vector in vitro was detected by Western blotting. In vivo, 18 Balb / c mice were selected and divided into 3 groups randomly, namely, vector + vector group ), Adr DNA vaccine + HBsAg protein vaccine, HBsAg protein vaccine + HBsAg protein vaccine; At the 0th week, the mice were immunized with vector, adr and HBsAg protein vaccine respectively, Mice were immunized with vector, HBsAg protein vaccine and HBsAg protein vaccine respectively in the fourth week. Serum HBsAg-specific antibodies were detected by enzyme-linked immunosorbent assay (ELISA), and HBsAg-specific IFN-γ secreting cells were detected by ELISPOT. Results: Adr + protein group and Protein + Protein group mice could express MHBs after transfection with 293T cells in vitro. Serum anti-HBs and Protein + Protein group had higher anti-HBs titer than adr + protein group, but there was no statistical significance. In the three groups, no specific IFN-γ secreted splenocytes were detected in the vector + vector group Adr + protein group and Protein + Protein group were all detected, and the number of adr + protein group-specific cells was significantly higher than that of protein + protein group (P <0.001). Conclusion: Pre-sensitization of Hepatitis B virus surface antigen (MHB) nucleic acid vaccine can significantly enhance the cellular immune response to hepatitis B virus (HBsAg) vaccine in Balb / c mice.