Mechanisms of probiotic inhibition of pathogenic oral bacteria

益生菌抑制口腔致病菌的机制

• 批准号: 9120044
• 负责人: Louis Cornacchione
• 金额: $ 4.01万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-03-04 至 2019-03-03
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9120044
• 关键词: Affect; Alveolar Bone Loss; Anions; Bacteria; Biochemical Genetics; Biological Assay; Cations; Celiac Disease; Chromatography; Clinical; Complement; Dairy Products; Data; Dental caries; Development; Disease; Escherichia coli; Food; Fractionation; Future; Gel Chromatography; Genes; Genetic Techniques; Genomics; Goals; Growth; Health; High-Throughput Nucleotide Sequencing; Immune; In Vitro; Individual; Inflammation; Inflammatory; Intestines; Irritable Bowel Syndrome; Knock-out; Laboratories; Lactobacillus; Lead; Left; Libraries; Location; Mass Spectrum Analysis; Massive Parallel Sequencing; Molecular Weight; Mouth Diseases; Mutagenesis; Oral; Oral Microbiology; Oral cavity; Organism; Periodontal Diseases; Periodontitis; Population; Porphyromonas gingivalis; Predisposition; Prevention strategy; Preventive therapy; Preventive treatment; Probiotics; Proteins; Recombinant Proteins; Recombinants; Research; Resistance; Streptococcus mitis; Streptococcus mutans; Techniques; Technology; Testing; Time; Tissues; Tooth Diseases; Tooth Loss; Tooth structure; Virulence; Work; Yogurt; base; career; clinical application; clinical efficacy; cost; cost effective; cytotoxicity; design; experience; inhibitor/antagonist; microorganism; mutant; novel; oral bacteria; pathogen; pathogenic bacteria; protective effect; protein purification; public health relevance; research study; scaling and root planing; skills; socioeconomics; standard of care; symptom treatment; treatment strategy

 DESCRIPTION (provided by applicant): Probiotics are microorganisms that when consumed confer a positive or beneficial effect on the host. Use of probiotics in the oral cavity as preventative therapies are just beginning to be explored for dental caries and periodontal disease. Porphyromonas gingivalis (Pg) is an important organism in the onset and progression of periodontal disease. Current treatment for Pg associated periodontitis is largely based on treatment of symptoms rather than preventative therapies. We have isolated several probiotic strains of Lactobacillus delbreuckii from dairy products that inhibit Pg-induced cytotoxicity and Pg growth in vitro. Furthermore, we have demonstrated that supernatants derived from these strains of L. delbreuckii inhibit Pg growth. Initial testing indicates that the inhibitory molecule appears to be a protein with a molecular weight between 30-100 kDa. We propose to isolate and identify the secreted factor from L. delbreuckii by anion/cation exchange chromatography and mass spectrometry. Using this data, we will identify the genomic location encoding the inhibitory factor. We will clone and purify a recombinant inhibitory factor to carry out further testing. Using a previously constructed transposon library in Pg, we will use massively parallel sequencing technology to quickly and quantitatively screen for genes in Pg that interact with the inhibitory supernatant and/or recombinant inhibitory protein to quickly identify potential mechanisms of action. Lastly, we will examine the susceptibility profile of clinical isolates of Pg and other oral bacteria to L. delbreuckii inhibitory supernatants and/or purified recombinant inhibitory protein in an effort to determine spectrum of action of the L. delbreuckii inhibitor against clinical Pg isolates. Completion of the aims outlined in this application may identify a novel interspecies inhibitory mechanism and demonstrate the feasibility of a clinical application for our L. delbreuckii probiotic strains and/or their secreted products.

 描述（由申请人提供）：益生菌是一种微生物，在食用时会对宿主产生积极或有益的影响。在口腔中使用益​​生菌作为龋齿和牙周病（Pg）的预防疗法才刚刚开始探索。 ）是牙周病发生和进展的重要生物体，目前对 Pg 相关牙周炎的治疗主要基于症状治疗而不是预防性治疗。乳制品中的几种德氏乳杆菌益生菌菌株可在体外抑制 Pg 诱导的细胞毒性和 Pg 生长。此外，我们还证明，来自这些德氏乳杆菌菌株的上清液可抑制 Pg 生长。 似乎是一种分子量在 30-100 kDa 之间的蛋白质。我们建议通过阴离子/阳离子交换色谱法和质谱法分离和鉴定德氏乳杆菌的分泌因子，我们将利用该数据确定编码该蛋白质的基因组位置。我们将克隆纯化一个重组抑制因子，利用之前在Pg中构建的转座子文库，利用大规模并行测序技术来快速定量筛选基因。在 Pg 中与抑制性上清液和/或重组抑制蛋白相互作用，以快速识别潜在的作用机制最后，我们将检查 Pg 临床分离株的敏感性概况。 和其他口腔细菌对德氏乳杆菌抑制性上清液和/或纯化的重组抑制蛋白的影响，以努力确定德氏乳杆菌抑制剂针对临床Pg分离株的作用谱。完成本申请中概述的目标可以鉴定新的种间抑制性。机制并证明我们的德氏乳杆菌益生菌菌株和/或其分泌产物的临床应用的可行性。