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CFP10:作为新型结核疫苗候选物的mFcγ2增强小鼠的免疫反应。

CFP10: mFcγ2 as a novel tuberculosis vaccine candidate increases immune response in mouse.

作者信息

Baghani Ali Asghar, Soleimanpour Saman, Farsiani Hadi, Mosavat Arman, Yousefi Masoud, Meshkat Zahra, Rezaee Seyed Abdolrahim, Jamehdar Saeid Amel, Eydgahi Mohammad Reza Akbari, Sadeghian Hamid, Ghazvini Kiarash

机构信息

Antimicrobial Resistance Research Center, Bu-Ali Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran; Department of Microbiology and Virology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.

HIV/AIDS, HTLV and Viral Hepatitis Research Center, Iranian Academic Center for Education, Culture and Research (ACECR), Mashhad, Iran.

出版信息

Iran J Basic Med Sci. 2017 Feb;20(2):122-130. doi: 10.22038/ijbms.2017.8231.

Abstract

OBJECTIVES

Despite treatment with antibiotics and vaccination with BCG, tuberculosis (TB) is still considered as one of the most important public health problems in the world. Therefore, designing and producing a more effective vaccine against TB seems urgently. In this study, immunogenicity of a fusion protein which consisting or comprising CFP-10 from Mycobacterium tuberculosis and the Fc-domain of mouse IgG2a was evaluated as a novel subunit vaccine candidate against TB.

MATERIALS AND METHODS

The genetic constructs were cloned in pPICZαA expression vector and recombinant vectors (pPICZαA-CFP-10: Fcγ2a and pPICZαA-CFP-10:His) were transformed into Pichia pastoris. To evaluate the expression of recombinant proteins, SDS-PAGE and immunoblotting were used. The immunogenicity of recombinant proteins, with and without BCG were assessed in BALB/c mice and specific cytokines against recombinant proteins (IFN-γ, IL-12, IL-4, IL-17 and TGF-β) were evaluated.

RESULTS

The levels of IFN-γ and IL-12 in mice that received recombinant proteins was higher than the control groups (BCG and PBS). Thus, both recombinant proteins (CFP-10:Fcγ2a and CFP-10:His) could excite good response in Th1-cells. The Fc-tagged protein had a stronger Th1 response with low levels of IL-4, as compared to CFP-10:His. However, the highest level of Th1 response was observed in groups that were vaccinated with BCG (prime) and then received recombinant protein CFP-10: Fcγ2a (booster).

CONCLUSION

The results demonstrated that binding mice Fc-domain to CFP-10 protein can increase the immunogenicity of the subunit vaccine. Further studies, might be able to design and produce a new generation of subunit vaccines based on the Fc-fused immunogen.

摘要

目的

尽管使用了抗生素治疗并接种了卡介苗,但结核病(TB)仍被视为世界上最重要的公共卫生问题之一。因此,迫切需要设计和生产一种更有效的抗结核疫苗。在本研究中,评估了一种由结核分枝杆菌的CFP-10和小鼠IgG2a的Fc结构域组成或包含的融合蛋白作为新型抗结核亚单位疫苗候选物的免疫原性。

材料与方法

将基因构建体克隆到pPICZαA表达载体中,并将重组载体(pPICZαA-CFP-10:Fcγ2a和pPICZαA-CFP-10:His)转化到巴斯德毕赤酵母中。为评估重组蛋白的表达,使用了SDS-PAGE和免疫印迹法。在BALB/c小鼠中评估了重组蛋白(有或无卡介苗)的免疫原性,并评估了针对重组蛋白的特异性细胞因子(IFN-γ、IL-12、IL-4、IL-17和TGF-β)。

结果

接受重组蛋白的小鼠中IFN-γ和IL-12水平高于对照组(卡介苗和PBS)。因此,两种重组蛋白(CFP-10:Fcγ2a和CFP-10:His)均可在Th1细胞中激发良好反应。与CFP-10:His相比,带有Fc标签的蛋白具有更强的Th1反应且IL-4水平较低。然而,在接种卡介苗(初免)然后接受重组蛋白CFP-10:Fcγ2a(加强免疫)的组中观察到最高水平的Th1反应。

结论

结果表明,将小鼠Fc结构域与CFP-10蛋白结合可提高亚单位疫苗的免疫原性。进一步的研究可能能够基于Fc融合免疫原设计和生产新一代亚单位疫苗。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/669a/5339651/81c45a41d366/IJBMS-20-122-g002.jpg

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