Targeting bacterial virulence as a novel antibiotic and vaccine approach

针对细菌毒力作为一种新型抗生素和疫苗方法

• 批准号: 1973726
• 金额: --
• 依托单位: Imperial College London
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2017
• 资助国家: 英国
• 起止时间: 2017 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-1973726
• 关键词: Targeting; bacterial; virulence; novel; antibiotic

Where a bacterial virulence factor such as SpyCEP is alone, sufficient to cause disease, that virulence factor represents a suitable target for both vaccine development and therapeutic approaches.Aim 1 Detailed characterization of the enzymatic activity of purified recombinant SpyCEPSpyCEP is known to cleave and inactivate CXCL8 and other ELR+ chemokines resulting in impaired neutrophil recruitment to the site of infection. It is likely that cleavage also impacts many other known CXCL8-mediated effects. Due to a prior lack of adequate quantities of purified enzyme, a systematic study of enzyme kinetics has not been previously undertaken, though would be required for development of any inhibitor or assay of inhibition. As such, at the start of this work I will undertake kinetic studies using purified SpyCEP enzyme from GAS supernatant and recombinantly produced enzyme (all available in the laboratory) in order to determine the KM and Vmax parameters of SpyCEP using ELISA to monitor CXCL8 cleavage and SDS-PAGE or MS-based analyses if necessary. SpyCEP appears to have a number of additional activities that may relate to previously unknown effects of CXCL8, or to additional previously unknown substrates. These should be clarified in order to more systematically assess the efficacy of any vaccine or therapeutic.

如果像SpyCEP这样的细菌毒力因子单独存在，足以引起疾病，那么该毒力因子代表了疫苗开发和治疗方法的合适靶点。已知spycep可裂解并灭活CXCL8和其他ELR+趋化因子，导致中性粒细胞向感染部位募集受损。裂解也可能影响许多其他已知的cxcl8介导的效应。由于先前缺乏足够数量的纯化酶，尽管开发任何抑制剂或抑制测定都需要对酶动力学进行系统的研究，但以前没有进行过。因此，在这项工作开始时，我将使用从GAS上清纯化的SpyCEP酶和重组生产的酶（都在实验室中可用）进行动力学研究，以确定SpyCEP的KM和Vmax参数，使用ELISA监测CXCL8裂解，必要时使用SDS-PAGE或MS-based分析。SpyCEP似乎具有许多额外的活性，这些活性可能与CXCL8先前未知的作用或其他先前未知的底物有关。应澄清这些问题，以便更系统地评估任何疫苗或治疗方法的功效。

项目成果

Extracellular bacterial lymphatic metastasis drives Streptococcus pyogenes systemic infection

• DOI: 10.1038/s41467-020-18454-0
• 发表时间: 2020-09-17
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Siggins，Matthew K.;Lynskey，Nicola N.;Sriskandan，Shiranee
• 通讯作者: Sriskandan，Shiranee