设计出显示多份结核病抗原的壶状病毒颗粒

IF 2.5 4区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Biotechnology and Bioprocess Engineering Pub Date : 2024-03-11 DOI:10.1007/s12257-024-00089-3
R. Princess, M. L. Stephen Raj
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摘要

激发抗体和细胞介导的免疫反应是成功开发结核病(TB)疫苗的关键。结核分枝杆菌(Mtb)抗原 CFP10 和 ESAT6 是经过验证的强效候选疫苗,需要适当的佐剂才能引发更好的免疫反应。携带重复拷贝的外来抗原的病毒样颗粒可诱导 T 细胞和 B 细胞介导的免疫反应,这种免疫反应是抵御细胞内病原体所必需的。在这项研究中,我们通过将源自约翰逊草花叶病毒的衣壳蛋白(CP)基因与 CFP 10 或/和 ESAT 6 基因进行翻译融合,开发出了表面显示分枝杆菌抗原的杂交壶状病毒样颗粒(PVLPs)。将携带融合构建体的重组质粒转化到大肠杆菌中,表达融合蛋白,即 ESAT6-CP、CP-CFP10 和 ESAT6-CP-CFP10,并在变性条件下使用 Ni-NTA2+ 亲和层析法进行纯化。通过逐渐去除变性条件,嵌合 CP 融合蛋白在体外自组装成 PVLP。纯化的携带Mtb抗原的混合PVLPs注射给小鼠后,与不使用任何佐剂免疫相同抗原相比,ESAT6和CFP10抗原的免疫原性均有所增强。用嵌合 PVLPs 免疫小鼠脾细胞的体外刺激可上调参与结核免疫反应的细胞因子的表达。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Engineering potyvirus-like particles to display multiple copies of tuberculosis antigens

Elicitation of antibody and cell-mediated immune responses are crucial for successful vaccine development against tuberculosis (TB). Mycobacterium tuberculosis (Mtb) antigens CFP10 and ESAT6, potent and proven vaccine candidates require appropriate adjuvants to trigger better immune response. Virus-like particles carrying repetitive copies of foreign antigens can induce both T and B cell-mediated immunity required for conferring protection against intracellular pathogens. In this study, we developed hybrid potyvirus-like particles (PVLPs) displaying mycobacterial antigens on their surface by translationally fusing the coat protein (CP) gene derived from Johnson grass mosaic virus with CFP 10 or/and ESAT 6 gene(s). The recombinant plasmids carrying fusion constructs were transformed into Escherichia coli, the fusion proteins, viz. ESAT6-CP, CP-CFP10 and ESAT6-CP-CFP10, were expressed and purified using Ni-NTA2+ affinity chromatography under denaturing conditions. The chimeric CP fusion proteins were self-assembled in vitro into PVLPs by the gradual removal of denaturing conditions. The purified hybrid PVLPs carrying Mtb antigens when injected into mice showed enhanced immunogenicity for both ESAT6 and CFP10 antigens compared to the same antigens immunized without any adjuvant. In vitro stimulation of splenocytes derived from mice immunized with chimeric PVLPs upregulates the expression of cytokines involved in TB immune response.

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来源期刊
Biotechnology and Bioprocess Engineering
Biotechnology and Bioprocess Engineering 工程技术-生物工程与应用微生物
CiteScore
5.00
自引率
12.50%
发文量
79
审稿时长
3 months
期刊介绍: Biotechnology and Bioprocess Engineering is an international bimonthly journal published by the Korean Society for Biotechnology and Bioengineering. BBE is devoted to the advancement in science and technology in the wide area of biotechnology, bioengineering, and (bio)medical engineering. This includes but is not limited to applied molecular and cell biology, engineered biocatalysis and biotransformation, metabolic engineering and systems biology, bioseparation and bioprocess engineering, cell culture technology, environmental and food biotechnology, pharmaceutics and biopharmaceutics, biomaterials engineering, nanobiotechnology, and biosensor and bioelectronics.
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