HAEMOPHILUS INFLUENZAE OTITIS MEDIA--PROTECTIVE IMMUNITY

非分型嗜血杆菌引起的中耳炎和其他疾病 流感病毒(NTHI)仍然是本地区儿童的重大健康问题 在这个国家和世界其他地方。努力开发针对儿童的疫苗 一些研究人员正在进行疾病预防工作。我们有 鉴定出一个高分子量外膜蛋白家族 非分型流感嗜血杆菌是宿主抗体的主要靶标 均为非典型嗜血杆菌急性中耳炎恢复期儿童 媒体和对疾病具有天然免疫力的正常成年人。在一个 为了进一步鉴定这些蛋白质，我们克隆、表达、 并对NTHI原型株的两个基因进行了测序，这两个基因编码HIGH 与人类血清识别的那些分子量相同的蛋白质。 这两个克隆基因的核苷酸序列数据和其他 免疫学和形态学数据表明，NTHI高、分子 重量蛋白是结构上和可能功能上的类似物 百日咳杆菌丝状血凝素蛋白。丝状 血凝素是一种重要的黏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发挥类似的职能作用。我们将在体外使用几种 体内技术，以进一步表征这些高分子量 蛋白质，并确定它们在细菌黏附中的作用和 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的杀菌作用 存在于人类血清中的活性。我们将制备抗纯化的抗血清 高相对分子质量蛋白及其抗血清的检测 功能检测，包括体外和动物杀菌检测 灰鼠中耳炎体内防护实验。主动型 用纯化的蛋白质进行的免疫实验也将在 动物模型。我们将评估这些高分子分子的贡献 用腺样体器官培养法研究重量蛋白对NTHI黏附的影响 模特。最后，我们将进一步考察NTHI与 高分子量蛋白质和丝状血凝素蛋白 几种百日咳杆菌特异性粘附法检测百日咳杆菌 化验。由此产生的信息不仅对 加强我们对NTHI生物学的了解，也可能是 努力开发预防感染的疫苗的重要性 由这些微生物引起的。

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.