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一种包含PBP2a和自溶素的新型抗耐甲氧西林金黄色葡萄球菌重组候选疫苗在实验小鼠中具有保护作用。

A novel recombinant vaccine candidate comprising PBP2a and autolysin against Methicillin Resistant Staphylococcus aureus confers protection in the experimental mice.

作者信息

Haghighat Setareh, Siadat Seyed Davar, Sorkhabadi Seyed Mehdi Rezayat, Sepahi Abbas Akhavan, Mahdavi Mehdi

机构信息

Department of Microbiology, Faculty of Advanced Sciences and Technology, Pharmaceutical Sciences Branch, Islamic Azad University (IAUPS), Tehran, Iran.

Department of Mycobacteriology & Pulmonary Research, Microbiology Research Center, Pasteur Institute of Iran, Tehran, Iran.

出版信息

Mol Immunol. 2017 Nov;91:1-7. doi: 10.1016/j.molimm.2017.08.013. Epub 2017 Aug 31.

Abstract

The methicillin-resistant Staphylococcus aureus infection is a hot topic area in microbiology research. Here a novel vaccine candidate consisting of recombinant PBP2a and autolysin proteins were used. The proteins over expressed in E.coli BL21 (DE3) cells, and purified by the Ni-NTA affinity column and conjugated using EDAC and ADH as a linker and spacer, respectively. To investigate the immunogenicity and protective effects of recombinant proteins, 5 and 20μg of proteins in various formulations were subcutaneously injected in different groups. Two booster vaccinations were carried out in three-week intervals and blood samples were collected three weeks after each injection. To evaluate the immune response, total IgG, IgG1, IgG2a, and IgG2b were analyzed. Immunization of mice with r-autolysin and r-autolysin-PBP2a mixture raised total IgGantibody. Additionally, both IgG1 and IgG2a responses induced. Opsonophagocytosis assay showed that anti r-PBP2a and r-autolysin IgG not only promoted phagocytosis of S.aureus, but also decreased the number of viable bacterial cells. Furthermore, survival rate of experimental mice increased in the bacteremia infection. Our results demonstrated that active vaccination with a mixture of r-PBP2a/r-autolysin and conjugate form vaccine reduced the mortality rate and protected mice against lethal MRSA challenge as well as single proteins.

摘要

耐甲氧西林金黄色葡萄球菌感染是微生物学研究中的一个热门领域。在此,使用了一种由重组PBP2a和自溶素蛋白组成的新型候选疫苗。这些蛋白在大肠杆菌BL21(DE3)细胞中过表达,通过Ni-NTA亲和柱纯化,并分别使用EDAC和ADH作为连接剂和间隔物进行偶联。为了研究重组蛋白的免疫原性和保护作用,将不同制剂中的5μg和20μg蛋白皮下注射到不同组中。每隔三周进行两次加强免疫,并在每次注射后三周采集血样。为了评估免疫反应,分析了总IgG、IgG1、IgG2a和IgG2b。用重组自溶素和重组自溶素-PBP2a混合物免疫小鼠可产生总IgG抗体。此外,还诱导了IgG1和IgG2a反应。调理吞噬试验表明,抗重组PBP2a和重组自溶素IgG不仅促进了金黄色葡萄球菌的吞噬作用,还减少了活菌数量。此外,实验小鼠在菌血症感染中的存活率提高。我们的结果表明,用重组PBP2a/重组自溶素混合物和偶联形式疫苗进行主动免疫可降低死亡率,并保护小鼠免受致命性耐甲氧西林金黄色葡萄球菌攻击,单一蛋白也有同样效果。

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