DNA疫苗预敏蛋白疫苗增强策略对乙型肝炎病毒表面抗原蛋白免疫应答的影响

发布时间：2018-01-28 23:12

本文关键词： 核酸疫苗乙型肝炎表面抗原中蛋白初免-加强免疫策略　出处：《现代生物医学进展》2009年06期 　论文类型：期刊论文

【摘要】：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(P0.001),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。
[Abstract]:Objective: to observe the presensitivity of nucleic acid vaccine. Effect of hepatitis B virus HBsAg protein vaccine enhanced immunization on immune response in Balb/c mice. Methods: Transfection. TM liposomes expressed hepatitis B virus surface antigen proteins (. The nucleic acid vaccine pSW3891/MHBs/ad(r is referred to as Adr. for short. And empty vector pSW3891 (vector) was transfected into 293T cells in vitro. Western blotting was used to detect the expression of Adr-vector in vitro. In vivo study, 18 Balb/c mice were selected, 6 mice in each group were randomly divided into 3 groups, that is, vector vector group. Adr nucleic acid vaccine HBsAg protein vaccine HBsAg protein vaccine HBsAg protein vaccine in ADR protein group and protein Protein group; Mice were immunized with vectoradr and HBsAg protein vaccine at week 0 and intramuscular injection with vectorus HBsAg protein vaccine at week 4. HBsAg protein vaccine was used to immunize mice. Elisa was used to detect the specific antibody to HBsAg in serum of mice. The HBsAg polypeptide specific IFN- 纬 secreting cells of mouse spleen cells were detected by enzyme-linked immunoblot assay (Elisa). Results 293T cells were transfected with 10% ADR in vitro. It can express the protein of hepatitis B virus surface antigen (HBsAg). The results of in vivo study showed that serum anti-HBs could be detected in all mice except vector vector group, ADR protein group and protein Protein group. The titer of anti-HBs in Protein Protein group was higher than that in adr protein group, but there was no statistical significance. No specific INF- 纬 secreted splenocytes were detected in the vector vector group, while in the adr protein group, no specific INF- 纬 secreted spleen cells were detected in the three groups. All the mice in Protein Protein group could be detected. The number of specific cells in adr protein group was significantly higher than that in protein protein group (P 0.001). Conclusion: hepatitis B virus surface antigen protein MHBsAg) nucleic acid vaccine presensitive. It can obviously enhance the cellular immune response of Balb/c mice to hepatitis B HBsAg protein vaccine.
【作者单位】：南京医科大学第一附属医院感染病科;美国马萨诸塞州大学医学院;
【基金】：国家自然科学基金面上项目(30371276) 135重点人才基金项目(135-044)
【分类号】：R392;R512.6
【正文快照】： 前言特异性细胞免疫功能低下是慢性乙型肝炎的主要致病机制之一[1],核酸疫苗能够刺激机体产生特异性免疫应答而具有潜在治疗价值,有动物实验结果表明,HBsAg核酸疫苗能刺激宿主产生一定强度的特异性免疫应答[2-4],但目前存在的一个明显问题是动物越大,DNA疫苗的免疫效力越低[5-

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