Human and mouse antibodies against influenza virus

描述(由申请人提供)：流感疫苗的效力各不相同，对老年人的保护作用往往较低。一种假设是，疫苗在老年人中的低效可能是由于流感特异性但非中和抗体的优势所致。这些非中和抗体可能是针对早期流感病毒，针对变性或内部病毒蛋白，或低亲和力。这项研究的长期目标是通过最大限度地诱导针对疫苗毒株的中和抗体，并最大限度地减少对变性糖蛋白、内部蛋白和早期病毒的反应，来开发更有效的流感疫苗。对多次接种或感染的受试者血清中和抗体和非中和抗体的定性和定量研究将在三个特定目标下进行。 目的1：反复接种流感疫苗或感染后，血清中有哪些抗体特异性？在疫苗接种者和感染者中检测针对H3N2型病毒主要抗原漂移变异的血清抗体，以及在针对这些病毒的单抗的竞争分析中，将提供关于中和抗体与非中和抗体、交叉反应抗体与毒株特异性抗体的定量信息，以及非中和抗体反应是否主要针对较老的病毒或变性病毒粒子。 目的2：在抗原漂移过程中，主要抗原区是否发生优势变化？多年来，血凝素(HA)上的主要抗原部位发生了相对优势的变化。将通过竞争分析来测量HA表位的相对免疫优势和抗体的相对亲和力，并通过构建重组HA基因来改进结果，重组HA基因只表达一个主要的抗原位点。 目的3：设计疫苗策略以优化中和抗体的生产。AIMS 1和AIMS 2的结果将提供对血清中非中和抗体来源的测量，并使疫苗战略得以发展，以最大限度地提高中和抗体。这些实验旨在填补人类抗体对流感疫苗反应广度的知识空白，并将这一知识应用于疫苗生产，以提高保护性中和抗体的比例。这可能对保护老年人免受流感侵袭特别有帮助。

DESCRIPTION (provided by applicant): Influenza vaccines have variable efficacy and protection is often low in the elderly. One hypothesis is that low effectiveness of vaccine in the elderly may be due to a preponderance of influenza-specific but non-neutralizing antibodies. These non-neutralizing antibodies may be against earlier influenza viruses, against denatured or internal viral proteins, or of low avidity. The long-term goal of this research is to develop more effective influenza vaccines by maximizing induction of neutralizing antibodies against the vaccine strain and minimizing the response to denatured glycoproteins, internal proteins, and earlier viruses. Qualitative and quantitative studies of neutralizing and non-neutralizing antibodies in sera of multiply-vaccinated or infected subjects will be undertaken in three Specific Aims. Aim 1: What antibody specificities are present in serum after repeated flu vaccination or infection? Measurement of serum antibodies in vaccinees and infected subjects against the major antigenic drift variants of H3N2 viruses, and in competition assays against monoclonal antibodies specific to those viruses will provide quantitative information on neutralizing versus non-neutralizing, cross-reactive versus strain-specific antibodies, and if the non-neutralizing antibody response is primarily directed against older viruses or against denatured virions. Aim 2: Do the major antigenic regions change in dominance during antigenic drift? The major antigenic sites on the hemagglutinin (HA) have changed in relative dominance over the years. The relative immunodominance of epitopes on the HA and the relative avidity of antibodies will be measured by competition assays, and the results refined by constructing recombinant HA genes engineered to express only one of the major antigenic sites. Aim 3: Devise a vaccine strategy to optimize production of neutralizing antibodies. The results of Aims 1 and 2 will provide a measure of the origins of non-neutralizing antibodies in serum, and allow development of a vaccine strategy to maximize neutralizing antibodies. These experiments are designed to fill a large gap in knowledge of the breadth of human antibody response to influenza vaccines, and to apply this knowledge to vaccine production to improve the ratio of protective, neutralizing antibodies. This may be of particular help in protecting the elderly from influenza.