HAEMOPHILUS INFLUENZAE OTITIS MEDIA--PROTECTIVE IMMUNITY

流感嗜血杆菌中耳炎--保护性免疫

• 批准号: 3131977
• 负责人: Stephen J. Barenkamp
• 金额: $ 10.39万
• 依托单位: WASHINGTON 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 1989-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3131977
• 关键词: Haemophilus influenzae; Haemophilus influenzae vaccines; active immunization; affinity chromatography; antibacterial antibody; antiserum; bacterial antigens; bactericidal immunity; cross immunity; enzyme linked immunosorbent assay; genetic strain; human subject; humoral immunity; immunofluorescence technique; immunological substance; microorganism immunology; monoclonal antibody; otitis media; passive immunization; radioimmunoassay; surface antigens

Acute otitis media is a persisting health problem among children in this country and elsewhere. Nontypable Haemophilus influenzae are a major cause of this illness. The objective of this proposal is to gain additional understanding of the host factors important in protective immunity against otitis media caused by this organism. In Aim I will define the importance of antibody to a 39 Kd major outer membrane protein of nontypable Haemophilus influenzae in mediating protective immunity in the chinchila otitis model. Both convalescent sera from animals recovered from infection and immune sera which are protective in passive protection experiments contain high levels of antibody against the 39 Kd protein of the homologous nontypable Haemophilus strain. Affinity-purified or nonclonal antibodies will be prepared against the 39 Kd proteins of several prototype strains and assayed in passive protection experiments. Purified 39 Kd proteins from prototype strains will be tested as vaccine candidates in the animal model. In Aim 2, I will define strain differences in the 39 Kd proteins of nontypable Haemophilus as they relate to immunity to otitis. Affinity-purified or monclonal antibody preparations recognizing the 39 Kd proteins will be tested against a panel of nontypable strains in fluorescent antibody, whole cell radioimmunoprecipitation and bactericidal assays. In vitro studies will be followed with preliminary in vivo cross-protection assays. In Aim 3, I will define the cell surface antigens of nontypable Haemophilus which stimulate serum and middle ear fluid antibody responses in the course of human infection. Acute and convalescent samples will be assayed with radioimmunoprecipitation, ELISA, and bactericidal assays. Absorption experiments will be performed with purified outer membrane proteins and lipopolysaccharide in an effort to identify targets of bactericidal antibody. The information derived from this work should permit us to better assess whether vaccination is a feasible strategy to pursue for the prevention of nontypable Haemophilus otitis media.

急性中耳炎是儿童持续存在的健康问题 国家和其他地方。 非嗜血型流感是主要原因 这种疾病。 该提议的目的是获得更多 了解宿主因素在保护性免疫中重要的因素 该生物引起的中耳炎。 目的我将定义重要性 抗39 kd的不可能剂外膜蛋白的抗体 流感嗜血杆菌在介导丁香的保护性免疫中 中耳炎模型。 两者都是从感染中回收的动物的康复血清 在被动保护实验中具有保护性的免疫血清 包含针对同源物的39 kD蛋白的高水平抗体 不可用的嗜血杆菌菌株。 亲和纯化或非克隆抗体 将针对几种原型菌株的39 kD蛋白制备 并在被动保护实验中进行测定。 纯化的39 kD蛋白 从原型菌株中将测试作为动物中的疫苗候选物 模型。 在AIM 2中，我将定义39 kD蛋白的应变差异 与中耳炎的免疫有关，它们是不可耕种的嗜血杆菌。 识别39 kD的亲和纯化或蒙克隆抗体制剂 蛋白质将针对一组不可替代的菌株进行测试 荧光抗体，全细胞放射免疫沉淀和杀菌 测定。 体外研究将进行体外研究 跨保护测定法。 在AIM 3中，我将定义细胞表面抗原 刺激血清和中耳液的不可用的嗜血杆菌 人类感染过程中的抗体反应。 急性和 康复样品将用放射免疫沉淀（ELISA）测定 和杀菌测定法。 将使用吸收实验 纯化的外膜蛋白和脂多糖 鉴定杀菌抗体的靶标。 从 这项工作应该使我们能够更好地评估疫苗接种是否是 可行的策略，以预防不可能的嗜血杆菌 耳炎培养基。