Human & mouse antibodies against influenza virus

描述(申请人提供)：该项目的长期目标是优化流感疫苗接种策略。三价亚单位流感疫苗是安全的，而且往往有效，但在老年人，他们最容易受到流感并发症的影响，或者当疫苗成分与正在传播的病毒不是很匹配时，效力就会降低。人们对人类产生的流感中和抗体的谱知之甚少。这项拟议的研究旨在定性和定量检测流感感染或疫苗诱导的个体抗体特异性，产生中和抗体的效率，以及这些抗体在抗原漂移中所起的作用。这个假设是，在老年人中，存在着大多数低特异性、中和能力差的抗体，这些抗体不仅提供了很差的保护，而且可能允许选择足够强大的抗原变体，以成为新的流行毒株。具体目标是： 目的1：确定杂交瘤分泌的单个人类抗体的中和能力，并将其与其表位位置及其筛选逃逸突变体的潜力相关联。 目的：比较青壮年和老年恢复期或免疫志愿者血清中抗流感抗体的性质。将筛选血清的结合和对新旧病毒的抑制活性，以及对一组重组血凝素和神经氨酸酶蛋白的结合和抑制活性，这些重组血凝素和神经氨酸酶蛋白中的抗原区通过替换新序列而被抑制。 目的：利用成熟的小鼠杂交瘤技术辅助人类抗体的研究。表位将通过逃逸突变分析和X射线结晶学进行绘制。

DESCRIPTION (provided by applicant): The long-term goal of this project is to optimize vaccination strategies against influenza. The trivalent subunit influenza vaccine is safe and often effective, but efficacy is reduced in the elderly, who are most susceptible to complications from influenza, or when the vaccine components are not a close match to the circulating viruses. Little is known about the spectra of influenza-neutralizing antibodies produced in humans. The proposed study is to qualitatively and quantitatively examine the antibody specificities induced in an individual by influenza infection or vaccine, the efficiency of producing neutralizing antibodies, and the role these play in antigenic drift. The hypothesis is that in the elderly there is a preponderance of low-specificity, poorly neutralizing antibodies that not only offer poor protection but may allow selection of antigenic variants that are sufficiently robust to become a new epidemic strain. The Specific Aims are: Aim 1: To determine the neutralizing power of individual human antibodies secreted by hybridomas and relate this to their epitope locations and their potential to select escape mutants. Aim 2: To compare the properties of anti-influenza antibodies present in sera of young adult and elderly convalescent or immunized volunteers. Sera will be screened for binding and for inhibitory activity against old and new viruses, and also against a panel of recombinant hemagglutinin and neuraminidase proteins in which antigenic regions are suppressed by substituting novel sequences. Aim 3: To use the well-established mouse hybridoma technology to assist the human antibody studies. Epitopes will be mapped by escape mutant analysis and x-ray crystallography.