BACTERIAL SURFACE PROTEINS: POTENTIAL TARGETS FOR SEPSIS

细菌表面蛋白：脓毒症的潜在目标

• 批准号: 6510047
• 负责人: Judith Hellman
• 金额: $ 12.46万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-06-01 至 2005-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6510047
• 关键词: active immunization; bacterial disease; bacterial proteins; bactericidal immunity; blood toxicology; clinical research; human subject; laboratory mouse; laboratory rat; lipopolysaccharides; membrane proteins; nonhuman therapy evaluation; pathologic process

Bacterial cell wall components released into the bloodstream are believed to be important in eliciting the inflammatory response that causes Gram-negative Sepsis. The bacterial outer membrane contains lipopolysaccharide (LPS), outer membrane proteins (OMPs), and phospholipids. LPS has been studied extensively as an important mediator in sepsis; the importance of OMPs in sepsis has not been systematically evaluated. Our preliminary data indicate that complexes containing three conserved OMPs and LPS are released from bacteria into human serum and are also released into the circulation of septic rats. We have identified the three proteins and have found that at least one of the proteins has biological activity. The major goals of this project are to further characterize release of the three OMPs in sepsis, to assess their roles in the pathogenesis of sepsis, and to evaluate anti-OMP immunity as a means of treating sepsis. The first specific aim is to assess release of OMPs and OMP/LPS complexes in humans with Gram- negative sepsis and in two models of experimental Gram-negative sepsis in rats. The second specific aim is to evaluate individual OMPs, OMP/LPS complexes, and OMPs on the surface of intact bacteria for biological activity in vitro on isolated immune cells, an in vivo by studying lethality, induction of cytokines in blood and organs, and pulmonary neutrophil sequestration in LPS resistant and LPS responsive mice. The third specific aim is to evaluate protective effects of active immunity on toxicity of OMP containing preparations in mice and in the rat infection models of sepsis. The applicant's long-term career objective is to become an independent investigator studying basic mechanisms of Gram- negative infection and sepsis. The career development plan is designed to expand the candidate's research knowledge base through hands-on research and educational activities. These will include course work (laboratory and didactic), attendance at and participation in laboratory meetings and research seminars, and formal and informal interactions with the Advisory Committee members. The ultimate goal of the candidate is to apply the fundamental knowledge of mechanisms involved in sepsis to develop anti-sepsis strategies that target bacterial toxins.

据信，释放到血液中的细菌细胞壁成分在引起引起革兰氏阴性败血症的炎症反应中很重要。 细菌外膜含有脂多糖（LPS），外膜蛋白（OMP）和磷脂。 LPS已被广泛研究为败血症的重要介体。 OMP在败血症中的重要性尚未系统地评估。 我们的初步数据表明，包含三个保守的OMP和LP的复合物从细菌释放到人血清中，也释放到化粪池的循环中。 我们已经确定了三种蛋白质，并发现其中至少一种蛋白质具有生物活性。该项目的主要目标是进一步表征败血症中三个OMP的释放，以评估它们在败血症的发病机理中的作用，并评估抗药免疫作为治疗败血症的一种手段。 第一个具体目的是评估具有革兰氏阴性败血症的人的OMP和OMP/LPS复合物的释放，以及在大鼠中的两个实验革兰氏阴性败血症模型中的释放。 第二个具体目的是评估完整细菌表面上的单个OMP，OMP/LPS复合物和OMPS，用于在分离的免疫细胞上进行生物学活性，这是通过研究致死性，血液和器官中的细胞因子诱导的体内，以及LPS抗LPS和LPS的肺中性粒细胞序列。 第三个具体目的是评估主动免疫对小鼠和败血症大鼠感染模型中含有OMP的毒性的保护作用。申请人的长期职业目标是成为一个独立研究者，研究革兰氏阴性感染和败血症的基本机制。 职业发展计划旨在通过动手研究和教育活动来扩大候选人的研究知识库。 这些将包括课程工作（实验室和教学），参加并参加实验室会议和研究研讨会以及与咨询委员会成员的正式和非正式互动。 候选人的最终目标是应用败血症涉及的机制的基本知识，以开发针对细菌毒素的抗盐策略。