CHILDREN'S ANTIBODIES TO POLYSACCHARIDES--V REGION GENES

产生了B细胞抗体库的显著多样性 通过不同免疫球蛋白组的组合分类 (Ig)生殖系元素 重排IG基因的进一步多样化 是由一种独特的体细胞超变过程提供的 个人 这些机制中的每一种对功能性抗体的贡献 剧目不详。 抗体对荚膜的反应 一种主要的儿童细菌病原体嗜血杆菌的多糖（PS 流感病毒B型（HiB）的抗体数量有限 种（clnotypes）。 因此，它应该作为一个很好的模式， 检查不同种系变量的相对贡献（V） 区域和体细胞突变对功能性免疫应答的影响。 此外，对这种感染的易感性是年龄的函数， 可能与V基因的个体发育有关。 这个项目的总体目标是 我们的建议是研究人类V区基因库的表达 作为年龄和抗原暴露形式的函数。 本研究 我们提出1）分离人抗HiB PS B细胞， 来自婴儿、儿童和成人的杂交瘤， 用Hib PS的T非依赖性（TI）和T依赖性（TD）形式免疫 2）通过序列分析确定IgV区库 杂交瘤cDNA克隆的克隆化和血清抗独特型分析 抗体和外周B细胞之间的关系，3）检查 V区序列或IG同种型，以及抗Hib PS的功能活性 检测IG基因在体细胞免疫中的作用 对TD和TI两者应答IgV区域多样性的超突变 疫苗。 B细胞杂交瘤抗体的V基因可以容易地被克隆。 测序 此外，开发V区专用 抗独特型抗体将允许分析大量的 血清样品，而不需要分离，克隆和测序一个大的 杂交瘤的数量。 比较在不同条件下捕获的V基因 免疫后的时间点将决定V 基因使用和体细胞突变对抗体应答成熟的影响。 确定体细胞突变对TI和TD反应的影响 将阐明T细胞在这一独特过程中的作用。 最后， 对不同年龄段V区使用情况的了解将决定个体发育 V基因的使用，并可能提供线索的易感性，某些 患者感染。

The remarkable diversity of the B cell antibody repertoire is generated through the combinatorial assortment of distinct groups of immunoglobulin (Ig) germline elements. Further diversification of rearranged Ig genes is provided by a unique process of somatic hypermutation. The individual contribution of each of these mechanisms to the functional antibody repertoire is unknown. The antibody response to the capsular polysaccharide (PS) of a major childhood bacterial pathogen, Haemophilus influenzae type b (Hib), is restricted to a limited number of antibody species (cl notypes). Therefore it should serve as an excellent model to examine the relative contribution of different germline variable (V) regions and somatic mutation to the functional immune response. Furthermore, susceptibility to this infection is a function of age and may be related to the ontogeny of V gene usage. The overall goal of this proposal is to study the expression of the human V region gene repertoire as a function of age and the form of antigenic exposure. In this study we propose 1) To isolate human antiHib PS B cells in the form of hybridomas from infants, children and adults at various times following immunization with T-independent (TI) and T-dependent (TD) forms of Hib PS vaccine, 2) To determine the IgV region repertoire by sequence analysis of cDNA clones of hybridomas and by anti-idiotype analysis of serum antibodies and peripheral B cells, 3) To examine the relationship between V region sequence or Ig isotype, and functional activity of anti-Hib PS antibodies, and 4) To examine the contribution of Ig gene somatic hypermutation to IgV region diversity in response to both TD and TI vaccines. The V genes of the B cell hybridoma antibodies can be readily sequenced. In addition, the development of V region specific anti-idiotype antibodies will allow the analysis of a large number of serum samples without the need to isolate, clone and sequence a large number of hybridomas. Comparison of V genes captured at different timepoints following immunization will determine the contribution of V gene usage and somatic mutation to maturation of the antibody response. Determining the impact of somatic mutation in both TI and TD responses will clarify the role of T cells in this unique process. Finally, knowledge of V region usage at different ages will determine the ontogeny of V gene usage and may provide clues to the susceptibility of certain patients to this infection.