HAEMOPHILUS INFLUENZAE OTITIS MEDIA--PROTECTIVE IMMUNITY

急性中耳炎是一个持续的健康问题，在儿童中， 国家和其他地方。 不定型的流感嗜血杆菌是一个主要原因 这种疾病。 该提案的目的是增加 了解保护性免疫中重要的宿主因素， 由这种微生物引起的中耳炎。 在目标中，我将定义 抗体的39 Kd的主要外膜蛋白的不可分型 流感嗜血杆菌介导灰鼠保护性免疫 耳炎模型。 来自感染恢复的动物的恢复期血清 以及在被动保护实验中具有保护性的免疫血清 含有高水平的抗39 Kd蛋白质的同源 非典型嗜血杆菌菌株。 亲和纯化或非克隆抗体 将针对几种原型菌株的39Kd蛋白制备 并在被动保护实验中测定。 纯化的39 Kd蛋白 将作为候选疫苗在动物身上进行测试 模型 在目标2中，我将定义39 Kd蛋白的菌株差异， 因为它们与耳炎的免疫有关。 识别39 Kd的亲和纯化或单克隆抗体制剂 蛋白质将针对一组无法分型的菌株进行测试， 荧光抗体、全细胞放射免疫沉淀和杀菌 测定。 体外研究之后将进行初步体内研究 交叉保护试验。 在目标3中，我将定义细胞表面抗原 刺激血清和中耳液的非典型嗜血杆菌 在人类感染过程中的抗体反应。 急性和 将用放射免疫沉淀，ELISA， 和杀菌测定。 吸收实验将使用 纯化的外膜蛋白和脂多糖， 鉴定杀菌抗体靶点。 导出的所述信息 这项工作应该使我们能够更好地评估疫苗接种是否是一种 预防非典型嗜血杆菌的可行策略 中耳炎

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.