VIRULENCE AND IMMUNITY TO MUCOID P. AERUGINOSA

描述：(改编自申请者摘要)：这项计划的主要目标 项目是了解毒力和宿主免疫力的基本方面 铜绿假单胞菌的粘液性菌株是发病率和 囊性纤维化患者的死亡率。大约2500名患有CF的新生儿 每年在世界各地出生，到青春期，其中80%的人将成为 慢性感染粘液性铜绿假单胞菌。临床数据清楚地表明， 肺功能下降和最终的早期死亡率发生在P。 铜绿假单胞菌感染的CF患者是由于粘液性铜绿假单胞菌感染和 不对任何其他病原体，包括非粘液性铜绿假单胞菌。因此， 预防新感染或治疗已确定的感染 针对粘液性铜绿假单胞菌的免疫治疗药物应该有一个主要的 对慢性阻塞性肺疾病患者健康的影响。要评估的主要假设是 慢性粘液性铜绿假单胞菌感染依赖于 粘液多糖(MEP)表面抗原的乙酰化版本，以及 针对MEP上的乙酰化表位的抗体提供了保护 慢性感染。这项工作的一个关键组成部分将利用我们的能力 启动和建立转基因铜绿假单胞菌慢性粘液性感染 Cf老鼠，到目前为止，其他使用这些老鼠的研究人员还没有意识到这一工具。 第一个假设将使用体外研究进行评估，以确定 乙酰化MEP对粘液型铜绿假单胞菌的抗药性 光声杀伤以及在CF小鼠体内的研究。在老鼠身上，我们会 评价不同菌株对甲基苯丙氨酸的乙酰化能力 建立和维持长期的殖民统治。第二个假设是 不同种类MEP结合疫苗的生产效果评价 乙酰化水平，通过化学操作制备。共轭关系 疫苗将在小鼠和兔子身上进行免疫原性评估，并对 它们在体外诱导抗体能力方面的特征 介导对大量粘液型铜绿假单胞菌菌株的光学杀灭。 接下来，疫苗(S)以最好的能力诱导出高滴度的 广泛反应的光学抗体将在转基因CF小鼠身上进行测试，使用 预防感染的主动疫苗接种和被动治疗 已确定的感染。此外，我们还将测试我们提出的假设 存在针对非乙酰化表位的预先存在的非光学抗体 铜绿假单胞菌粘液性感染发病前患者MEP的研究 干扰了安装保护性的光学响应的能力 乙酰化表位。最后，我们将评估最近一些 制备抗MEP的人源性单抗，以降低粘液P的水平。 感染CF小鼠肺组织中的铜绿假单胞菌。这项工作的结果应该是 进一步了解粘液型铜绿假单胞菌感染的发病机制， 并推动有效疫苗和被动治疗试剂的开发 目的：预防和治疗CF患者的粘液性铜绿假单胞菌感染。

DESCRIPTION: (Adapted from applicant's abstract): The major goals of this project are to understand basic aspects of virulence and host immunity to mucoid strains of P. aeruginosa that are major causes of morbidity and mortality in cystic fibrosis (CF) patients. About 2500 new babies with CF are born each year worldwide, and by adolescence 80 per cent of them will become chronically infected with mucoid P. aeruginosa. Clinical data clearly show that the decline in pulmonary function and ultimate early morality that occurs in P. aeruginosa-infected CF patients is due to mucoid P. aeruginosa infection and not to any other pathogen, including non-mucoid P. aeruginosa. Therefore, preventing new infections or treating established infections with immunotherapeutic agents that target mucoid P. aeruginosa should have a major impact on the health of CF patients. The primary hypotheses to be evaluated are that chronic mucoid P. aeruginosa infection is dependent on elaboration of an acetylated version of the mucoid exopolysaccharide (MEP) surface antigen, and that antibodies to the acetylated epitopes on MEP confer protection from chronic infection. A key component of this work will use our ability to initiate and establish a chronic mucoid P. aeruginosa infection in transgenic CF mice, a tool that, to date has eluded other researchers using these mice. The first hypothesis will be evaluated using in vitro studies to ascertain the ability of the acetylated MEP to confer resistance on mucoid P. aeruginosa to opsonic killing as well as in in vivo studies in CF mice. In the mice we will evaluate the ability of strains with differing abilities to acetylate MEP to establish and maintain a chronic colonization. The second hypothesis will be evaluated by production of conjugate vaccines composed of MEP with differing levels of acetylation, prepared by chemical manipulations. The conjugate vaccines will be evaluated for their immunogenicity in mice and rabbits and for their in vitro characteristics in regard to their ability to elicit antibodies that mediate opsonic killing of a multitude of mucoid P. aeruginosa strains. Next the vaccine(s) with the best ability to elicit high titers of broadly-reactive opsonic antibody will be tested in transgenic CF mice, using both active vaccination to prevent9 infection and passive therapy of established infection. In addition, we will test our proposed hypothesis that the presence of pre-existing, non-opsonic antibodies to non-acetylated epitopes on MEP in CF patients prior to the onset of mucoid P. aeruginosa infection interferes with the ability to mount a protective, opsonic response to the acetylated epitopes. Finally, we will evaluate the potential of some recently prepared human monoclonal antibodies to MEP to reduce the levels of mucoid P. aeruginosa in the lungs of infected CF mice. The results of this work should further our insights into the pathogenesis of mucoid P. aeruginosa infection, and promote development of effective vaccines and passive therapeutic reagents to prevent and treat mucoid P. aeruginosa infections in CF patients.