HUMAN ANTIBODY REPERTOIRE TO H INFLUENZAE B

描述(改编自申请者《S摘要》)：研究 这份提案中描述的内容代表了我们继续努力 描述可变区(V)的遗传和结构基础 编码针对b型病毒的人类抗体(Ab)谱系 (Hib)囊多糖(PS)，并了解其动力学。 这个保护性的抗体曲目。不同的Hib-PS结合疫苗 在婴儿中诱导Hib PS特异性抗体 抗Hib的功能活性，这些差异与 特定独特型/可变区(V)的使用。此外，V 曲目修改是由于婴儿年龄增长而发生的 成年人不同地利用特定的抗Hib PS区。这个 研究人员的研究试图定义分子机制 这些现象背后的原因。这一提议的一个核心假设是 由不同Hib可变驱动的抗体V区的超突变 PS疫苗和暴露于自然产生的抗原作为 衰老的后果，在决定这两个V 区域/独特型表达和抗体功能性质。要定义和 操纵婴儿和成人的抗Hib PS V区曲目， 将使用两种实验方法，独特型分析和 人源抗体Fab片段在噬菌体组合中的表达 图书馆。一种广谱的抗Hib PS V区基因序列分析 将在从接种疫苗的婴儿身上获得的B淋巴细胞上进行 成年人。这种方法将使我们能够确定V基因元件 编码婴儿抗Hib PS谱系，到目前为止一直是 分子定义无法获得的。此外，这一分析将 允许评估V区突变的程度 由不同的疫苗和老化过程引起的。血清学 将制备与新的独特型/V区起反应的试剂，并 将用于扩展我们对V区域表达的分析 由疫苗类型和年龄定义的人群。两国之间的关系 还将研究Hib-PS的自身抗体反应性和特异性。 由于V基因片段中使用的抗Hib PS抗体也很常见 由抗自我抗体使用。通过利用噬菌体操纵V基因成分 组合文库、V区重组实验可以 以评估单个V区的特异性潜力， 重链V区和轻链V区在其中的相对作用 威慑抗体的特异性和功能，以及结构决定因素 自我和非自我反应性。亲和力和特异性分析将 在序列定义的、噬菌体衍生的Fabs上执行以关联V使用 和功能。本周的长期目标是理解 塑造人类抗体曲目的进化和躯体力量 PS抗原。勾勒出管理继承的规则， 发育调控及其表达的分子基础 这些重要的抗体特异性将帮助我们理解 不同的PS疫苗产生的抗体有明显的差异 功能能力，因此将使合理设计 未来用于婴儿的更有效的PS疫苗以及 老年人。

DESCRIPTION (Adapted from the applicant"s abstract): The research described in this proposal represents a continuation of our efforts to delineate the genetic and structural bases of the variable (V) regions encoding the human antibody (Ab) repertoire specific for the type b (Hib) capsular polysaccharide (PS), and to understand the dynamics of this protective Ab repertoire. The different Hib PS conjugate vaccines induce Hib PS-specific Ab in infants that differ profoundly in functional activity against Hib, and these differences correlate with usage of particular idiotypes/variable (V) regions. In addition, V repertoire modification occurs as a consequence of aging since infants and adults differentially utilize particular anti-Hib PS regions. The investigator's research seeks to define the molecular mechanisms underlying these phenomena. A central hypothesis of this proposal is that hyper mutation of Ab V regions, variably driven by different Hib PS vaccines and by exposure to naturally-occurring antigens as a consequence of aging, plays a central role in determining both V region/idiotypic expression and Ab functional quality. To define and manipulate the anti-Hib PS V region repertoires of infants and adults, two experimental methodologies will be used, idiotypic analysis and expression of human Ab Fab fragments in bacteriophage combinatorial libraries. An extensive anti-Hib PS V region cDNA sequence analysis will be performed on B lymphocytes obtained from vaccinated infants and adults. This approach will allow us to determine the V gene elements encoding the infant anti-Hib PS repertoire, which heretofore have been inaccessible to molecular definition. In addition, this analysis will permit assessment of the extent to which V region mutation has been induced by different vaccines and by the aging process. Serological reagents reactive with new idiotypes/V regions will be prepared and they will be used to extend our analysis of V region expression in populations defined by vaccine type and age. The relationship between autoantibody reactivity and specificity for Hib PS will also be studied since the V gene segments used anti-Hib PS antibodies also are commonly used by anti-self Ab. By manipulating V gene components using phage combinatorial libraries, V region recombination experiments can be performed to assess the specificity potential of individual V regions, the relative roles that the heavy and light chain V regions play in deterring Ab specificity and function, and structural determinants of self and non-self reactivity. Avidity and specificity analyses will be performed on sequence-defined, phage-derived Fabs to correlate V usage and function. The long term objectives of this week are to understand the evolutionary and somatic forces that shape human Ab repertoires to PS antigens. Delineating the rules governing the inheritance, the developmental control and the molecular basis for the expression of these important Ab specificities will help us to understand how different PS vaccines generate antibodies with such markedly different functional capabilities, and thus will enable the rational design of more effective future PS vaccines for use in infants as well as the elderly.