A Chlamydomonas-derived Human Papillomavirus 16 E7 vaccine induces specific tumor protection.
A Chlamydomonas-derived Human Papillomavirus 16 E7 vaccine induces specific tumor protection.
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DOI：
10.1371/journal.pone.0061473
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Giuliano G
中科院分区：
文献类型：
--
作者：
Demurtas OC;Massa S;Ferrante P;Venuti A;Franconi R;Giuliano G
人乳头瘤病毒 (HPV) 16 型的 E7 蛋白参与恶性细胞转化，是开发针对 HPV 相关病变和癌症的治疗性疫苗的关键抗原。这种疫苗抗原以活性形式重组生产并符合良好生产规范 (GMP) 对于开发有效的疫苗起着至关重要的作用。在临床前模型中，植物生产的基于 E7 的治疗性疫苗已被证明对肿瘤消退和保护具有活性。然而，全植物疫苗生产的一些缺点鼓励我们探索在莱茵衣藻中生产基于 E7 的治疗性疫苗，莱茵衣藻是一种易于生长和转化且完全符合 GMP 指南的生物体。在莱茵衣藻叶绿体 psbD 5' UTR 和 psbA 3' UTR 的控制下，单独或作为与亲和标签（His6 或 FLAG）的融合物编码 E7GGG（E7 癌蛋白的突变、减毒形式）的表达盒通过同源重组引入莱茵衣藻叶绿体基因组中。蛋白质大部分为可溶性，占可溶性蛋白质总量的0.12%。对两种标记形式优化并进行了亲和纯化。在接种总衣藻提取物和亲和纯化蛋白的 C57BL/6 小鼠中检测到特异性抗 E7 IgG 和 E7 特异性 T 细胞增殖的诱导。用表达 E7 蛋白的肿瘤细胞系攻击后，实现了高水平的肿瘤保护。莱茵衣藻叶绿体是用于生产可溶性免疫原性形式的 E7GGG 蛋白的合适表达系统。在封闭和无菌条件下的生产凸显了微藻作为生产人类疫苗替代平台的潜力。
The E7 protein of the Human Papillomavirus (HPV) type 16, being involved in malignant cellular transformation, represents a key antigen for developing therapeutic vaccines against HPV-related lesions and cancers. Recombinant production of this vaccine antigen in an active form and in compliance with good manufacturing practices (GMP) plays a crucial role for developing effective vaccines. E7-based therapeutic vaccines produced in plants have been shown to be active in tumor regression and protection in pre-clinical models. However, some drawbacks of in whole-plant vaccine production encouraged us to explore the production of the E7-based therapeutic vaccine in Chlamydomonas reinhardtii, an organism easy to grow and transform and fully amenable to GMP guidelines. An expression cassette encoding E7GGG, a mutated, attenuated form of the E7 oncoprotein, alone or as a fusion with affinity tags (His6 or FLAG), under the control of the C. reinhardtii chloroplast psbD 5′ UTR and the psbA 3′ UTR, was introduced into the C. reinhardtii chloroplast genome by homologous recombination. The protein was mostly soluble and reached 0.12% of total soluble proteins. Affinity purification was optimized and performed for both tagged forms. Induction of specific anti-E7 IgGs and E7-specific T-cell proliferation were detected in C57BL/6 mice vaccinated with total Chlamydomonas extract and with affinity-purified protein. High levels of tumor protection were achieved after challenge with a tumor cell line expressing the E7 protein. The C. reinhardtii chloroplast is a suitable expression system for the production of the E7GGG protein, in a soluble, immunogenic form. The production in contained and sterile conditions highlights the potential of microalgae as alternative platforms for the production of vaccines for human uses.
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DOI：
10.1038/nbt1088-1204
发表时间：
1988-10-01
期刊：
BIO-TECHNOLOGY
影响因子：
--
作者：
HOPP, TP;PRICKETT, KS;CONLON, PJ
通讯作者：
CONLON, PJ
影响因子：
5.4
作者：
FELTKAMP, MCW;SMITS, HL;KAST, WM
通讯作者：
KAST, WM
影响因子：
11.4
作者：
DEBUCHY, R;PURTON, S;ROCHAIX, JD
通讯作者：
ROCHAIX, JD
影响因子：
2.9
作者：
Alonso, LG;García-Alai, MM;de Prat-Gay, G
通讯作者：
de Prat-Gay, G
影响因子：
5.5
作者：
Massa, Silvia;Franconi, Rosella;Venuti, Aldo
通讯作者：
Venuti, Aldo