The Preparation of Human Papillomavirus Type 58 Vaccine and Exploring Its Biological Activity and Immunogenicity In Vitro
The Preparation of Human Papillomavirus Type 58 Vaccine and Exploring Its Biological Activity and Immunogenicity In Vitro
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人乳头瘤病毒58型疫苗的制备及其体外生物活性和免疫原性探讨
DOI：
10.1097/igc.0b013e31821dc8e5
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发表时间：
2011-04
影响因子：
4.8
通讯作者：
Jianfeng Zhou
中科院分区：
文献类型：
--
作者：
Dongrui Deng;Shixuan Wang;Li Meng;Ding Ma;shujie liao (廖书杰)(并列第一);Xiangyang Bai;Wei Wang;Ni Sima;Qian Xu;Tao Zhu;Gang Xu;Jianfeng Zhou
目的：人乳头瘤病毒（HPV）58 型是中国女性中第二常见的病毒感染。为了开发兼具预防和治疗功能的 HPV58 疫苗，我们生成了一种嵌合病毒样颗粒 (cVLP)。方法：cVLP 包含全长 L1 和 E7 氨基酸 (aa) 50 至 aa72 或 E7 aa4 至 aa12 的部分 E7 肽。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳显示 HPV58 L1-E7aa50-72 和 L1-E7aa4-12 融合蛋白，并通过蛋白质印迹进行确认（补充数字内容 1，http://links.lww.com/IGC/A40，其显示 HPV58 L1-E7aa50-72/4-12 与标准序列之间的蛋白质序列比对）。通过透射电子显微镜鉴定蛋白质折叠和cVLP的位置。采用酶联免疫斑点法检测融合蛋白的免疫原性。结果透射电镜显示融合蛋白自组装形成cVLPs，大部分颗粒位于sf-9昆虫细胞核内。 cVLPs表现出很强的凝集红细胞的能力，这与亲代VLPs不同。此外，纯化的HPV58 L1-E7aa50-72或L1-E7aa4-12融合蛋白诱导显着数量的表达干扰素γ的E7aa50-72或E7aa4-12特异性CD8+T细胞。我们的结果表明，在L1后面插入E7aa50-72或E7aa4-12肽不会破坏cVLP的组装，并提供有效的免疫原性和生物活性，这为进一步制备具有预防和治疗作用的HPV58疫苗用于治疗包括宫颈癌在内的HPV58相关疾病奠定了有力的基础。
Objectives: Human papillomavirus (HPV) type 58 is the second most prevalent virus infection among Chinese women. To develop an HPV58 vaccine that combines both prophylactic and therapeutic functions, we generate a chimeric virus-like particle (cVLP). Methods: The cVLPs contain both whole length L1 and parts of E7 peptides either from E7 amino acids (aa) 50 to aa72 or from E7 aa4 to aa12. The HPV58 L1-E7aa50-72 and L1-E7aa4-12 fusion proteins were revealed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and confirmed by Western blot (Supplementary Digital Content 1, http://links.lww.com/IGC/A40, which shows alignment of the protein sequence between HPV58 L1-E7aa50-72/4-12 and standard sequence). Protein folding and location of cVLPs were identified by transmission electron microscope. The immunogenicity of the fusion protein was tested by enzyme-linked immunospot assay. Results Transmission electron microscope showed that the fusion protein formed cVLPs by self-assembly and the majority of particles located in the nucleus of the sf-9 insect cells. The cVLPs displayed a strong ability to agglutinate erythrocytes, which is distinguished from the parental VLPs. In addition, the purified HPV58 L1-E7aa50-72 or L1-E7aa4-12 fusion protein induced significant numbers of interferon &ggr;-expressing E7aa50-72- or E7aa4-12-specific CD8+ T cells. Discussion Our results indicate that the insertion of the E7aa50-72 or E7aa4-12 peptides behind L1 did not disrupt the assembly of cVLPs and provided potent immunogenicity and bioactivity, which created a powerful basis for further preparations of HPV58 vaccines with prophylactic and therapeutic effects for the treatment of HPV58-related diseases including cervical cancer.
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DOI：
10.1002/9783527609314.ch5
发表时间：
1996
期刊：
Biochimica et biophysica acta
影响因子：
--
作者：
H. zur Hausen
通讯作者：
H. zur Hausen
DOI：
10.1099/0022-1317-75-7-1647
发表时间：
1994-07
期刊：
The Journal of general virology
影响因子：
--
作者：
Linda A. Selvey;Linda A. Dunn;R. Tindle;D. Park;Ian H. Frazer
通讯作者：
Linda A. Selvey;Linda A. Dunn;R. Tindle;D. Park;Ian H. Frazer
DOI：
--
发表时间：
2001-11
期刊：
Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology
影响因子：
--
作者：
A. Giuliano;M. Papenfuss;M. Abrahamsen;C. Denman;J. D. Zapien;J. L. N. Henze;L. Ortega;E. M. B. D. Galaz;Jennifer Stephan;Janine D. Feng;S. Baldwin;F. Garcia;K. Hatch
通讯作者：
A. Giuliano;M. Papenfuss;M. Abrahamsen;C. Denman;J. D. Zapien;J. L. N. Henze;L. Ortega;E. M. B. D. Galaz;Jennifer Stephan;Janine D. Feng;S. Baldwin;F. Garcia;K. Hatch
DOI：
10.1099/0022-1317-82-11-2799
发表时间：
2001-11
期刊：
The Journal of general virology
影响因子：
--
作者：
Slupetzky K;Shafti-Keramat S;Lenz P;Brandt S;Grassauer A;Sara M;Kirnbauer R
通讯作者：
Kirnbauer R
影响因子：
254.7
作者：
Parkin, DM;Bray, F;Pisani, P
通讯作者：
Pisani, P