AIMS:This study aimed to investigate the ability of CpG7909 adjuvant to enhance immunogenicity and protective efficacy of a subunit vaccine composed of ESAT6-Ag85A fusion protein (Pe685a) of Mycobacterium tuberculosis. METHODS AND RESULTS:ELISA was used to detect specific antibody and IFN-γ expression in sera; ELISPOT, to detect IFN-γ expression in splenocytes; MTT assay and FACS, to detect T-lymphocytes proliferation in spleens; and RT-PCR, to detect cytokines expression in lungs of mice after immunization. Bacterial load and histopathological lesions in lungs or spleens of mice challenged with Myco. tuberculosis H37Rv strain were analysed. Compared with incomplete Freund's adjuvant, CpG7909 induced more potent production of Pe685a-specific IgG2a/IgG1 antibody and higher expression of IFN-γ in sera, stimulated more generation of antigen-specific IFN-γ-secreting splenocytes, enhanced frequencies of CD3(+) CD4(+) and CD3(+) CD8(+) T-lymphocytes in spleen and increased transcription of TNF-α, IFN-γ, IL-6 and TLR9 in lung. However, lower bacterial load in lung and less severe lung pathology were not observed in CpG7909 group mice. CONCLUSIONS:CpG7909 is able to enhance immunological effects of Pe685a subunit vaccine, but does not confer significant protective efficacy against Myco. tuberculosis infection. SIGNIFICANCE AND IMPACT OF THE STUDY:CpG7909 as an adjuvant of subunit vaccine against Myco. tuberculosis is worthy of further investigation.

译文

目的:本研究旨在研究CpG7909佐剂增强由结核分枝杆菌的ESAT6-Ag85A融合蛋白(Pe685a)组成的亚单位疫苗的免疫原性和保护功效的能力。
方法与结果:采用ELISA法检测血清中的特异性抗体和IFN-γ表达。 ELISPOT,用于检测脾细胞中的IFN-γ表达; MTT法和FACS,检测脾脏中T淋巴细胞的增殖;和RT-PCR,以检测免疫后小鼠肺中的细胞因子表达。用Myco攻击的小鼠肺或脾脏中的细菌负荷和组织病理学损害。分析了结核H37Rv株。与不完全弗氏佐剂相比,CpG7909诱导更有效地产生Pe685a特异性IgG2a / IgG1抗体,并在血清中表达更高的IFN-γ,刺激更多抗原分泌IFN-γ的脾细胞生成,增加CD3()CD4的频率()和CD3()CD8()脾脏中的T淋巴细胞,并增加肺中TNF-α,IFN-γ,IL-6和TLR9的转录。但是,在CpG7909组小鼠中未观察到较低的肺部细菌负荷和较轻的肺部病理学。
结论:CpG7909能够增强Pe685a亚基疫苗的免疫学作用,但对Myco却没有明显的保护作用。结核感染。
该研究的意义和影响:CpG7909作为针对Myco的亚单位疫苗的佐剂。结核病值得进一步研究。

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