HAEMOPHILUS INFLUENZAE OTITIS MEDIA--PROTECTIVE IMMUNITY

流感嗜血杆菌中耳炎--保护性免疫

• 批准号: 3131983
• 负责人: Stephen J. Barenkamp
• 金额: $ 16.23万
• 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/3131983
• 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.

由非典型嗜血杆菌引起的中耳炎和其他疾病 流感病毒（NTHI）仍然是这一地区儿童的重大健康问题， 国家和世界其他地方。 为发展艾滋病毒/艾滋病疫苗所作的努力 许多研究人员正在研究预防疾病的方法。 我们有 鉴定了一个高分子量外膜蛋白家族 非典型流感嗜血杆菌是宿主抗体的主要靶标 这两种情况都发生在非典型嗜血杆菌急性中耳炎康复期的儿童中， 媒体和正常成年人谁是自然免疫的疾病。 中 为了进一步表征这些蛋白质，我们克隆，表达， 并对来自NTHI原型菌株的两个基因进行了测序， 分子量与人血清识别的蛋白质相同。 来自这两个克隆基因的核苷酸序列数据和另外的 免疫学和形态学数据表明，NTHI高分子， 重量蛋白质是结构和可能的功能类似物， 百日咳杆菌丝状血凝素蛋白。 丝状 血凝素是一种重要的粘附因子和保护性抗原， 百日咳杆菌 我们的假设是高分子量 为NTHI提供类似的功能。 我们将在体外使用几种药物， 进一步表征这些高分子量 蛋白质，并确定其在细菌粘附中的作用， 针对它们的抗体在宿主保护中。 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的杀菌剂 活性存在于人血清中。 我们将制备抗血清， 高分子量蛋白质，并测定产生的抗血清， 功能测定包括体外和动物的杀菌测定 在体灰鼠中耳炎保护实验。 活性 用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 使用腺样体器官培养物测定蛋白质对NTHI粘附的影响 模型 最后，我们将进一步研究NTHI与 高分子量蛋白和丝状血凝素蛋白 使用几种百日咳杆菌特异性粘附的百日咳杆菌 测定。 由此产生的信息不仅对以下方面有价值： 提高我们对NTHI生物学的理解， 努力开发预防感染疫苗的重要性 由这些生物体引起的。