人乳头瘤病毒58型治疗性复合DNA疫苗载体的构建  

Construction and identification of PVAX1-HPV58mE6E7Fc-hIL12 composite DNA vaccine vector

作　　者：王鹤[1] 于继云[2] 李力[1] 

机构地区：[1]广西医科大学附属肿瘤医院,南宁530021 [2]军事医学科学院基础医学研究所,北京100850

出　　处：《中国免疫学杂志》2012年第12期1110-1114,共5页Chinese Journal of Immunology

基　　金：2009年广西卫生厅重点课题资助项目(重200970)

摘　　要：目的:构建和表达人乳头瘤病毒(Human papilloma virus,HPV)58型相关宫颈癌治疗用PVAX1-HPV58mE6E7Fc-hIL12复合DNA疫苗载体。方法:将HPV58mE6E7融合基因片段插入真核表达载体pCI-Fc-GPI中,构建重组真核表达质粒pCI-sig-HPV58mE6E7-Fc-GPI。再将得到的sig-HPV58mE6E7-Fc-GPI片段插入新型疫苗载体PVAX1-IRES-hIL12中,构建PVAX1-HPV58mE6E7Fc-hIL12复合DNA疫苗载体,流式细胞术、免疫荧光及ELISA分别检测该疫苗载体的表达。结果:经限制性内切酶鉴定及测序分析证实PVAX1-HPV58mE6E7Fc-hIL12复合DNA疫苗载体构建成功。流式细胞术、免疫荧光及ELISA检测表明该疫苗载体中的融合抗原sig-HPV58mE6E7-Fc-GPI及分子佐剂人白介素12(hIL12)能够同时实现真核表达。结论:PVAX1-HPV58mE6E7Fc-hIL12复合DNA疫苗载体可作为治疗HPV58相关肿瘤及其癌前病变的候选疫苗载体。Objective：To construct and express a composite DNA vaccine vector named PVAX1-HPV58mE6E7Fc-hIL12 for human papilomavirus 58（HPV58）-associated cervical cancer.Methods：HPV58mE6E7 fusion gene was inserted into pCI-Fc-GPI eukaryotic expression vector,constructing a recombinant plasmid named pCI-sig-HPV58mE6E7-Fc-GPI.The fragment of sig-HPV58mE6E7-Fc-GPI was inserted into the novel vaccine vector PVAX1-IRES-hIL12,constructing PVAX1-HPV58mE6E7Fc-hIL12 composite DNA vaccine vector.Flow cytometry,immunofluorescence and ELISA were used to detect expression of the vaccine.Results：PVAX1-HPV58mE6E7Fc-hIL12 vaccine vector was successfully constructed and identified by restriction endonuclease and sequencing analysis.The expression analysis showed that fusion antigen sig-HPV58mE6E7-Fc-GPI and molecular adjuvant human interleukin 12（hIL12） can be realized at the same time.Conclusion：PVAX1-HPV58mE6E7Fc-hIL12 can be taken as a candidate of therapeutic composite DNA vaccine vector for HPV58-associated tumors and their precancerous transformations.

关 键 词：人乳头瘤病毒58型 E6基因 E7基因 DNA疫苗载体 真核表达 

分 类 号：R737[医药卫生—肿瘤] R34[医药卫生—临床医学]