HAEMOPHILUS INFLUENZAE OTITIS MEDIA--PROTECTIVE IMMUNITY

急性中耳炎是儿童长期存在的健康问题。 国家和其他地方。不能定型的流感嗜血杆菌是主要原因 关于这种疾病的。这项提议的目的是获得更多的 对保护性免疫中重要的宿主因素的认识 由这种微生物引起的中耳炎。在AIM中，我将定义 抗39kD主要外膜蛋白抗体的研究 流感嗜血杆菌介导金龟子的保护性免疫 中耳炎模型。从感染中恢复的动物的恢复期血清 和在被动保护实验中具有保护性的免疫血清 含有针对同源的39kD蛋白的高水平抗体 不可分型的嗜血杆菌菌株。亲和纯化抗体或非克隆抗体 将针对几个原型菌株的39kD蛋白进行准备 并在被动防护实验中进行了检测。纯化的39kD蛋白 原型菌株将作为候选疫苗在动物身上进行测试 模特。在目标2中，我将定义猪瘟病毒39kD蛋白的菌株差异。 非分型嗜血杆菌，因为它们与中耳炎的免疫力有关。 识别39kD的亲和纯化或单抗制剂 蛋白质将在一组非分型菌株中进行测试 荧光抗体、全细胞放射免疫沉淀与杀菌 化验。在体外研究之后，将进行初步的体内研究 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分型嗜血杆菌 人类感染过程中的抗体反应。急性和 恢复期样本将用放射免疫沉淀、酶联免疫吸附试验、 和杀菌试验。吸收实验将在以下条件下进行 纯化的外膜蛋白和脂多糖，努力 确定杀菌抗体的靶点。派生自 这项工作应该使我们能够更好地评估接种疫苗是否是一种 寻求预防非分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.