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学科主题: 感染疾病科
题名:
HBsAg 及其与小、鼠白介素-18融合蛋白表达质粒的构建和DNA免疫
其他题名: DNA-mediated immunization to hepatitis B surface antigen: effects of a plasmid coexpressing hepatitis B surface antigen and interleukin-18
作者: 柯亨宁; 斯崇文; 成军; 于敏; 刘丹
关键词: DNA免疫 ; 肝炎表面抗原,乙型 ; 白细胞介素18
刊名: 中华传染病杂志
发表日期: 2001
DOI: 10.3760/j.issn:1000-6680.2001.02.003
卷: 19, 期:2, 页:77-80
收录类别: 中国科技核心期刊 ; 中文核心期刊 ; CSCD
文章类型: Journal Article
摘要: 目的 观察乙型肝炎病毒表面抗原(HBsAg)基因表达质粒(pS)及其与鼠白细胞介素-18基因融合表达质粒(p18S)的构建及其诱导BALB/c鼠免疫应答的能力。方法 用聚合酶链反应(PCR)法获得的S基因克隆到真核表达质粒pcDNA3 1+上,构建pS。另用三步PCR法将小鼠白细胞介素-18基因融合到S基因的5′端,再插入到pcDNA3 1+的EcoRI位点上,构建了p18S。分别免疫10只和6只BALB/c小鼠,阴性对照用pcDNA3 1+免疫4只BALB/c小鼠。用酶联免疫方法 检测每份血清的抗-HBs的效价。采用3H标记法检测脾细胞HBsAg特异性细胞毒反应。结果 pS质粒免疫BALB/c小鼠多数于4周时出现抗-HBs,最高可达530 mIU/ml,平均为135 mIU/ml。p18S质粒也能诱导低水平的抗-HBs.平均为20 mIU/ml。pS和p18S诱导的HBsAg特异性的细胞毒活性分别为37 1%和34%,而pcDNA3.1+载体质粒免疫小鼠血清未能检出抗-HBs,细胞毒活性为13 2%。结论 pS质粒能够有效地诱导BALB/c小鼠的体液和细胞免疫反应,白细胞介素-18基因与S基因融合表达质粒免疫似乎不能增强免疫应答,相反机体免疫应答下降。 Objective To observe the specific humoral and cellular immune response in BALB/c mice injected with pS and p18S. Methods pS and p18S were constructed separately by inserting HBsAg gene fragment and the fusion gene fragment of HBsAg and mouse interleukin 18(IL-18) into the reading frame of pcDNA3. I + . Mice were injected with either plasmid intramuscularly in a total dose of 300 tμg per mouse. Every serum ~mple was detected for anti-HBs using enzyme linked immunosorbent assay(EL ISA).Furthermore, HBsAg specific cytotoxic T lymphocytes activity was measured. Results The expression of HBsAg was demonstrated by ELISA in p815 cells transfected with pS and p18S. pS can stimulate a positive antibody response. The average level was 135 mIU/ml, with the highest level of 530 mIU/ml. p18S could elicit relatively lower antibody response which was 20 mIU/ml. HBsAg specific CTL activities were 37.1%and 34% separately in pS and p18S immunized mice. It was only 13.2% when detected in pcDNA3.1 +immunization. Conclusion pS is effective to stimulate a humoral and cellular response in H-2d mice. IL-18gene can not enhance the immune response when fused with HBsAg gene. Conversely, it seems to inhibit an immune response.
语种: 中文
原文出处: 查看原文
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内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/41684
Appears in Collections:北京大学第一临床医学院_感染疾病科_期刊论文

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作者单位: 北京大学第一医院感染疾病科
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