[1]王亚婷,李文生,闫小飞,等.人乳头瘤病毒58型L1/E7“假病毒”疫苗的制备及免疫效果评价[J].现代检验医学杂志,2018,33(03):35-37，43.[doi:10.3969/j.issn.1671-7414.2018.03.011]
　WANG Ya-ting,LI Wen-sheng,YAN Xiao-fei,et al.Construction of Pseudovirions of Human Papillomavirus Type58 L1/E7 and Evaluation of Its Immunity Protective Efficacy[J].Journal of Modern Laboratory Medicine,2018,33(03):35-37，43.[doi:10.3969/j.issn.1671-7414.2018.03.011]
点击复制

人乳头瘤病毒58型L1/E7“假病毒”疫苗的制备及免疫效果评价()
分享到：

《现代检验医学杂志》[ISSN:/CN:]

卷:
第33卷
期数:
2018年03期
页码:
35-37，43
栏目:
论著
出版日期:
2018-07-17

文章信息/Info

Title:
Construction of Pseudovirions of Human Papillomavirus Type58 L1/E7 and Evaluation of Its Immunity Protective Efficacy
文章编号:
1671-7414(2018)03-035-04
作者:
王亚婷1; 李文生1; 闫小飞2; 齐宗利1; 郭 华1
1.陕西省人民医院病理科,西安 710068; 2.西安交通大学基础医学院,西安 710061
Author(s):
WANG Ya-ting1; LI Wen-sheng1; YAN Xiao-fei2; QI Zong-li1; GUO Hua1
1.Department of Pathology,Shaanxi Provincial People's Hospital,Xi'an 710068,China; 2.Basic Medical College of Xi'an Jiaotong University,Xi'an 710061,China
关键词:
人乳头瘤病毒58型;  病毒样颗粒;  假病毒;  细胞毒性T淋巴细胞
分类号:
R373; R392-33
DOI:
10.3969/j.issn.1671-7414.2018.03.011
文献标志码:
A
摘要:
目的 构建人乳头瘤病毒58型L1/E7“假病毒”疫苗,并研究其免疫学特性。方法 采用sf9昆虫-杆状病毒表达系统表达HPV58L1蛋白,体外解离重组病毒颗粒,向已经解离的衣壳中加入mE7-pcDNA3.1+质粒,CaCl2室温透析促使病毒样颗粒(VLP)重新聚合,用“假病毒” 颗粒滴鼻免疫小鼠,检测小鼠血清IgG,IgA中和抗体以及脾淋巴细胞IFN-γ的分泌水平和细胞毒性T淋巴细胞(CTL)反应。结果 SDS-PAGE,Western blot和红细胞凝集实验证实sf9昆虫-杆状病毒表达系统有效表达有功能活性的HPV58 L1蛋白,HPV58 L1蛋白能够有效的折叠、解聚以及再折叠为VLP。小鼠免疫保护实验发现,用包含有mE7-pcDNA3.1+质粒的HPV58 L1“假病毒”免疫小鼠,抗HPV58L1特异性IgG,IgA 明显升高(P<0.001),“假病毒”免疫组产生的IFN-γ明显高于VLP免疫组(P<0.001); “假病毒”免疫过的小鼠脾淋巴细胞可以特异性杀伤Tc-1细胞,引起CTL反应。结论 成功制备了人乳头瘤病毒58型L1/E7“假病毒”疫苗,能诱发免疫动物较强的体液免疫反应,激发保护性抗体的产生。
Abstract:
Abstract:Objective To develop a Human papilloma virus(HPV)type 58 L1/E7 pseudovirions vaccine and evaluate its immunity efficacy.Methods HPV58L1 protein was expressed in sf9 insect cell by using baculovirus expression system,after HPV58 VLPs was dialyzed with dissociation buffer,those were mixed with mE7-pcDNA3.1+ plasmid,reassembled into anovel psedovirions of HPV58.The ability of the reassembled into virus-like particles was identified by transmission electron microscope,and murine erythrocyte hemagglutination assay.In addition,HPV58 L1/E7 pseudovirions were immunized Balb/c mice by dripping nasal method,HPV58 L1 specific IgGs and IgAs antibody were examed by ELISA assay,and the concentration of IFN-γ were detected by ELISA method,cytotoxic T lymphocyte reaction of mouse spleen lymphocytes to TC-1 cells were detected by LDH method.Results HPV58 L1protein was highly expressed in sf9 insect system,the results showed that the expression of HPV58 L1 protein could induce the aggregation of red blood cells inmice,demonstrating the expression of HPV58 L1 showed good biological activity.After the CaCl2 dialysis,the dissociated virus like particles could be resynthesized into pseudovirions.HPV58 L1 specific IgGs and IgAs were significantly increased in the serum of Balb/c mice immunized with HPV58 VLP group and pseudovirions group(P<0.001).The serum levels of IFN-γ were significantly increased in the pseudovirions and VLP group(P<0.001),the spleen lymphocyte of pseudovirions immunized mice could specifically kill TC-1 cells,causing cytotoxic T lymphocyte reaction.ConclusionThe HPV58 L1/E7 psedovirions vaccine was been successfully constructed,and “pseudovirus” vaccine can induce a strong humoral immune response in immune animals and stimulate the production of protective antibodies.

参考文献/References:

[1] Bosch FX,Lorincz A,Mu(¨overn)oz N,et al.The causal relationbetween human papillomavirus and cervical cancer[J].J Clin Pathol,2002,55(4):244-265.
[2] 刘宝印,李 洁,De Villiers EM,等.我国人乳头瘤病毒58型感染与宫颈癌的关系[J].中华实验和临床病毒学杂志,1996,10(2):118-121. Liu BY,Li J.De villiers EM,et al.Investigation on relationship between the papillomavirus type 58(HPV58)infection and cervix cancer in China[J].Chinese J Exp Clin Virol,1996,10(2):118-121.
[3] 刘景会,刘玉林,褚 迪,等.截短型人乳头瘤病毒58型L1蛋白在昆虫细胞中的表达[J].中国生物制品学杂志,2015,3(28):233-238,244. Liu JH,Liu YL,Chu D,et al.Expression of truncated human papilomavirus 58 L1 protein in insect cells[J].Chinese Journal of Biologicals,2015,28(3):233-238,244.
[4] Mohsen MO,Zha L,Cabral-Miranda G,et al.Major findings and recent advances in virus-like particle(VLP)-based vaccines[J].Semin Immunol,2017,34:123-132.
[5] Roden RB,Weissinger EM,Henderson DW,et al.Ne-utralization of bovinepapillomavirus by antibodies to L1 and L2 capsid proteins[J].J Virol,1994,68(11):7570-7574.
[6] McIntyre MC,Frattini MG,Grossman SR,et al.Human papillomavirus type 18 E7 protein requires intact Cys-X-X-Cys motifs for zinc binding,dimerization,and transformation but not for Rb binding[J].J Virol,1993,67(6):3142-3150.
[7] Duensing S,Munger K.The human papillomavirus ty-pe 16 E6 and E7 oncoproteins independently induce numerical and structural chromosome instability[J].Cancer Res,2002,62(23):7075-7082.

备注/Memo

备注/Memo:
基金项目:陕西省科技攻关课题(2010K12-02),陕西省卫生计生科研项目(2016D035)。 作者简介:王亚婷(1979-),女,医学硕士,主要从事病理学及肿瘤生物学研究。 通讯作者:李文生,男,医学博士,主任医师,研究方向:肿瘤病理与肿瘤生物学,E-mail:liwensheng263@hotmail.com。
更新日期/Last Update: 2018-04-16