HPV18 L1基因重组DNA构建及其免疫小鼠体内特异抗体的测定  

Construction of HPV18 L1 gene recombinant plasmid and the detection of its specific antibody in immune mice

作　　者：贾士东[1] 谷鸿喜[1] 王文阁[2] 钟照华[1] 郭淑元[1] 凌虹[1] 贺福初[2] 

机构地区：[1]哈尔滨医科大学微生物学教研室,哈尔滨150086 [2]中国军事医学科学院实验血液学研究室,北京100850

出　　处：《中国免疫学杂志》2000年第7期356-358,共3页Chinese Journal of Immunology

基　　金：黑龙江省科委重点攻关项目!C98C0803

摘　　要：目的：人类乳头瘤病毒18型（HPV18）感染与宫颈癌的发生高度相关。其晚期基因L1编码的L1结构蛋白是刺激机体产生中和抗体的主要抗原。利用HPVL1基因重组质粒进行DNA疫苗的初步免疫学实验研究。方法：从HPV18－pBR322质粒中扩增HPV18L1基因，测序证明扩增片段的可靠性，构建了两个真核表达重组质粒HPV18L1-pcDNA3和HPV18L1－peEP4。将提纯的质粒DNA直接免疫小鼠，用HPV18L1抗原经ELISA检测小鼠血清相应抗体。结果：HPV18L1-pcDNA3和HPV18L1-pcEP4质粒DNA均能在小鼠体内表达并诱导产生HPV18特异性抗体。结论：HPV18L1重组质粒DNA直接注射小鼠可以刺激机体产生抗体。Objective: Infection of human papillomavirus type 18 is highly related to the development of cervical careinoma. The L1 protein which coded by the late gene L1 of HPV18 is the main antigen which induces the neutralization antibody. Method: The HPV18 L1 gene was amplified from pBR322-HPV18. The amplified DNA had been confirmed by sequencing. Two recombinant plasmids, peDNA3-HPV18L1 and pCEP4-HPV18L1 were constructed for expressing in mammals. BALB/C mice were immunolized by intramuscular injection with the purified plasmid DNAs. ELISA was employed for detection of HPV18 L1 antibody. Result: Results showed that HPV18 Specific antibody could be detected from the mice which had been injected with recombinant plasmid DNAs. Conclusion: The experiment indicated that immunization of both HPV18 L1-pcDNA3 and HPV18 L1-pCEP4 elicited neutralizing antibodies, and HPV18 L1-pCEP4 induced higher antibody titre.

关 键 词：晚期基因L1 基因表达 DNA HPV18 宫颈癌 

分 类 号：R737.33[医药卫生—肿瘤] R392.1[医药卫生—临床医学]