Conjugation Reaction with 8-Arm PEG Markedly Improves the Immunogenicity of Mycobacterium tuberculosis CFP10-TB10.4 Fusion Protein
Conjugation Reaction with 8-Arm PEG Markedly Improves the Immunogenicity of Mycobacterium tuberculosis CFP10-TB10.4 Fusion Protein
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8臂PEG缀合反应显着提高结核分枝杆菌CFP10-TB10.4融合蛋白的免疫原性
DOI：
10.1021/acs.bioconjchem.7b00131
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发表时间：
2017-06-01
影响因子：
4.7
通讯作者：
Hu, Tao
中科院分区：
文献类型：
--
作者：
Sun, Xiaowei;Yu, Weili;Hu, Tao
结核分枝杆菌 (Mtb) 是导致结核病 (TB) 的严重致命病原体。对于针对结核分枝杆菌感染的免疫保护而言，非常需要有效的疫苗接种。 CFP10和TB10.4是两种重要的免疫显性结核分枝杆菌分泌蛋白抗原，但其免疫原性较差。因此，需要抗原递送系统和佐剂来提高两种蛋白的免疫原性。本研究中使用 CFP10-TB10.4 融合蛋白 (CT) 作为抗原。将 4-6 个 CT 分子与 8 臂聚乙二醇 (PEG) 缀合在一个实体中，作为抗原递送系统。铝-洛索立宾混合物 (A-L) 和聚 (I:C) 作为佐剂。与CT相比，聚合CT（CT-PEG）在BALB/c小鼠中引起显着更高的CT特异性IgG滴度、更高的Th1和Th2型细胞因子以及更高的CD4（+）和CD4+ IFN-gamma（+）IL-4（+）细胞百分比。 A-L 和聚 (I:C) 的存在都可以增强对 CT-PEG 的免疫反应。与 8 臂 PEG 的缀合反应显示出提高 CT 免疫原性的主要驱动力。 SD 大鼠的药代动力学研究表明，与 8 臂 PEG 的缀合反应延长了 CT 的全身循环和对免疫系统的暴露。具有 A-L 的 CT PEG 对器官没有表现出明显的毒性，而具有聚 (I:C) 的 CT-PEG 对器官显示出一定的毒性。因此，通过Mtb分泌的蛋白抗原与8臂PEG的缀合反应以及随后添加A-L，可以合理地设计一种有效且安全的针对Mtb感染的疫苗。
Mycobacterium tuberculosis (Mtb) is a serious fatal pathogen responsible for tuberculosis (TB). Effective vaccination is highly desired for immunoprotection against Mtb infection. CFP10 and TB10.4 are two important immunodominant Mtb-secreted protein antigens, which suffer from poor immunogenicity. Thus, an antigen delivery system and adjuvants are needed to improve the immunogenicity of the two proteins. A CFP10-TB10.4 fusion protein (CT) was used as the antigen in the present study. Conjugation of 4-6 CT molecules in one entity with 8-arm polyethylene glycol (PEG) acted as an antigen delivery system. Aluminum-loxoribine mixture (A-L) and poly(I:C) functioned as the adjuvants. As compared with CT, the polymerized CT (CT-PEG) elicited significantly higher CT-specific IgG titers, higher Th1- and Th2-type cytokines and higher percentages of CD4(+) and CD4+ IFN-gamma(+) IL-4(+) cells in BALB/c mice. The presence of A-L and poly(I:C) could both increase the immune response to CT-PEG. Conjugation reaction with 8-arm PEG showed a predominant driving force to improve the immunogenicity of CT. Pharmacokinetic study in SD rats revealed that conjugation reaction with 8-arm PEG prolonged the systemic circulation of CT and exposure to the immune system. CT PEG with A-L showed no apparent toxicity to organs, whereas CT-PEG with poly(I:C) displayed some toxicity to organs. Thus, an effective and safe vaccine against Mtb infection could be rationally designed by conjugation reaction of Mtb-secreted protein antigen with 8-arm PEG and subsequent addition of A-L.