HAEMOPHILUS INFLUENZAE OTITIS MEDIA--PROTECTIVE IMMUNITY

流感嗜血杆菌中耳炎--保护性免疫

• 批准号: 3131980
• 负责人: Stephen J. Barenkamp
• 金额: $ 16.43万
• 依托单位: SAINT LOUIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-09-01 至 1995-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3131980
• 关键词: Bordetella pertussis; Haemophilus influenzae; active immunization; antibacterial antibody; bacterial antigens; bacterial proteins; bactericidal immunity; chinchilla; disease /disorder model; genetic strain; hemagglutinin; host organism interaction; human tissue; organ culture; otitis media

Otitis media and other illnesses caused by nontypable Haemophilus influenzae (NTHI) remain significant health problems for children in this country and elsewhere in the world. Efforts to develop vaccines for the prevention of disease are being made by a number of investigators. We have identified a family of high molecular weight outer membrane proteins nontypable Haemophilus influenzae which are major targets of host antibody both in children convalescing from nontypable Haemophilus acute otitis media and in normal adults who are naturally immune to disease. In an effort to further characterize these proteins, we have cloned, expressed, and sequenced two genes from a prototype NTHI strain which encode high molecular weight proteins identical to those recognized by the human sera. The nucleotide sequence data from these two cloned genes and additional immunologic and morphologic data suggest that the NTHI high, molecular weight proteins are structural and possibly functional analogues of the filamentous hemagglutinin protein of Bordetella Pertussis. Filamentous hemagglutinin is an important adherence factor and protective antigen of Bordetella pertussis. Our hypothesis is that the high molecular weight serve a similar functional role for NTHI. We will use several in vitro and in vivo techniques to further characterize these high molecular weight proteins and define their role in bacterial adhesion and the role of antibody directed against them in host protection. Specifically, we will perform absorption experiments to determine the contribution of antibody directed against the high molecular weight proteins to the bactericidal activity present in human sera. We will prepare antisera against purified high molecular weight proteins and assay the resulting antisera in functional assays which include bactericidal assays in vitro and animal protection experiments in the chinchilla otitis media in vivo. Active immunization experiments with purified proteins will also be performed in the animal model. We will assess the contribution of these high molecular weight proteins to the adherence of NTHI using an adenoidal organ culture model. Finally, we will further examine the relationship between the NTHI high molecular weight proteins and the filamentous hemagglutinin protein of Bordetella pertussis using several Bordetella pertussis specific adherence assays. The resulting information should be of value not only for enhancing our understanding of the biology of NTHI and but also may be of importance in efforts to develop vaccines protective against infection caused by these organisms.

中耳炎和其他由不可分型嗜血杆菌引起的疾病 流感病毒（NTHI）仍然是儿童面临的重大健康问题 国家和世界其他地方。 努力开发疫苗 许多研究人员正在开展疾病预防工作。 我们有 鉴定出一系列高分子量外膜蛋白 不可分型流感嗜血杆菌是宿主抗体的主要靶标 两名儿童均因非典型嗜血杆菌急性中耳炎而康复 媒体和对疾病自然免疫的正常成年人。 在一个 为了进一步表征这些蛋白质，我们已经克隆、表达、 并对来自原型 NTHI 菌株的两个基因进行了测序，这些基因编码高 与人类血清识别的分子量相同的蛋白质。 来自这两个克隆基因和其他基因的核苷酸序列数据 免疫学和形态学数据表明 NTHI 高分子 重量蛋白是结构和可能的功能类似物 百日咳博德特氏菌的丝状血凝素蛋白。 丝状 血凝素是重要的粘附因子和保护性抗原 百日咳博德特氏菌。 我们的假设是高分子量 为 NTHI 发挥类似的功能作用。 我们将使用几种体外和 体内技术进一步表征这些高分子量 蛋白质并定义它们在细菌粘附中的作用以及 在宿主保护中针对它们的抗体。 具体来说，我们将 进行吸收实验以确定抗体的贡献 针对高分子量蛋白质进行杀菌 人类血清中存在的活性。 我们将准备纯化的抗血清 高分子量蛋白质并测定所得抗血清 功能测定，包括体外和动物杀菌测定 对龙猫中耳炎的体内保护实验。 积极的 纯化蛋白质的免疫实验也将在 动物模型。 我们将评估这些高分子的贡献 使用腺样器官培养物测定蛋白质重量对 NTHI 粘附的影响 模型。 最后，我们进一步考察NTHI之间的关系 高分子量蛋白质和丝状血凝素蛋白质 百日咳博德特氏菌使用几种百日咳博德特氏菌特异性粘附 化验。 由此产生的信息不仅应该具有价值 增强我们对 NTHI 生物学的理解，但也可能是 开发预防感染的疫苗的重要性 由这些生物体引起。