COMPARATIVE ONTOGENETIC APPROACH TO VACCINE STRATEGY

疫苗策略的比较个体遗传学方法

• 批准号: 2076818
• 负责人: Max Dale Cooper
• 金额: $ 24.21万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-30 至 2000-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2076818
• 关键词: B lymphocyte; antigen receptors; biological signal transduction; cell age; cell differentiation; cell sorting; clone cells; cytokine; gene rearrangement; genetic transcription; hematopoietic stem cells; human tissue; immunogenetics; immunoglobulin genes; immunoglobulin isotypes; immunoglobulins; immunologic memory; interferon alpha; interferon gamma; laboratory mouse; laboratory rabbit; monoclonal antibody; polymerase chain reaction; vaccines

This project focuses on two major aspects of B cell production and function in humans: virgin and memory B cell generation. The differentiation steps that hemopoietic progenitor cells must undergo to give rise to clonally diverse virgin B lymphocytes will be analyzed in these experiments with an eye toward possible clinical manipulation of B cell production. Studies are proposed to dissect the (i) precise order of B-lineage differentiation events, (ii) order of transcriptional activity of genes important in this progression and (iii) assembly, expression and function of antigen receptor units as a function of differentiation stage and age. Fresh leads suggesting that B cell production in the bone marrow can be regulated both positively and negatively via non-antigen-specific receptors (IL-7, IFNalpha/Beta and CDI9) will be explored in one series of experiments. The hypothesis that B cells can be generated by an alternative differentiation pathway, involving light chain gene rearrangement before the rearrangement of conventional heavy chain genes, will be tested. In these experiments, (i) 'surrogate' heavy chains (MC) that couple kappa light chains to the Igalpha/beta signal transduction units on two B-lineage cell lines will be isolated and characterized, (ii) the surrogate RC gene cloned, sequenced, and expressed, (iii) monoclonal antibodies produced against the surrogate HC, and (iv) these reagents used as probes to identify bone marrow cells in this pathway. Studies will be conducted to pursue preliminary data suggesting the antibody repertoire may differ in B cells generated via this alternative pathway. The phenotypic and functional characteristics of memory B cells will be examined as a function of age. In these experiments IgA1 and IgA2 B cells will be purified from adult and cord blood samples for analysis of their (i) cell surface antigenic profile and activation status, (ii) mechanism of the isotype switch, (iii) antibody repertoire, (iv) expression of developmentally-regulated genes, and (v) functional capabilities. These studies of virgin IgM B cells and their IgA1 and IgA2 memory B cell progeny should provide valuable information for the design of vaccine strategy in individuals of various ages.

该项目着重于B细胞生产和生产的两个主要方面 人类的功能：处女和记忆B细胞的产生。这个 造血祖细胞必须经历的分化步骤 导致克隆性多样化的处女B淋巴细胞将在 这些实验着眼于可能的临床操作B 细胞生产。建议进行研究，以剖析(I)准确的顺序 B-谱系分化事件，(Ii)转录活性的顺序 在这一进程中重要的基因以及(Iii)组装、表达和 抗原受体单位的功能与分化阶段的关系 和年龄。新的线索表明骨髓中B细胞的产生 可以通过非抗原特异性的方式进行积极和消极的调节 受体(IL-7、IFNpha/Beta和CDI9)将在一系列 实验。假设B细胞可以由一种 另一种分化途径，涉及轻链基因 在常规重链基因重排之前的重排， 将会受到考验。在这些实验中，(I)“替代”重链(MC) 将kappa轻链偶联到免疫球蛋白α/β信号转导 两个B系细胞系上的单位将被分离和鉴定，(Ii) 代用RC基因的克隆、测序和表达，(Iii)单克隆 产生针对代用HC的抗体，以及(Iv)使用这些试剂 作为识别这一途径中的骨髓细胞的探针。研究将会是 进行的目的是寻找初步数据，表明抗体谱系 可能在通过这种替代途径产生的B细胞上有所不同。这个 记忆B细胞的表型和功能特征将是 作为年龄的函数进行检查。在这些实验中，IgA1和IgA2 B细胞 将从成人和脐带血样本中提纯，用于分析其 (I)细胞表面抗原谱和激活状态；(Ii)机制 同型转换，(Iii)抗体库，(Iv)表达 发育调节基因，以及(V)功能能力。这些 原始免疫球蛋白B细胞及其免疫球蛋白A_1和免疫球蛋白A_2记忆B细胞的研究 子代应为疫苗的设计提供有价值的信息 在不同年龄段的个人中的策略。