REGULATION OF IGE ANTIBODY PRODUCTION IN VITRO

IGE 抗体体外生产的调控

• 批准号: 3445716
• 负责人: Swey-Shen Chen
• 金额: $ 5.57万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-01-01 至 1987-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3445716
• 关键词: B lymphocyte; T lymphocyte; antibody formation; immunoglobulin E; immunoregulation

Antibody production of the immunoglobulin E (IgE) class is causally related to the allergic disorder in both human and experimental animals. Our understanding of this class of immunity has been greatly expanded in the past two decades through the elucidation of structure and function of IgE molecules. Recently, we have discovered a novel model system to induce polyclonal IgE tolerance in mice neonatally treated with IgE ligands. However, much remains to be learned about how regulatory cells exert control over the stage-specific differentiation of B-cells of the EgE lineage (BEpsilon-cells). This particular difficulty is largely due to the lack of a reproducible in vitro IgE antibody system. The objective of this research proposal is to determine the fine tolerogenic epitope inherent in the IgE molecule and examine how class-specific negative cells influence the stage-specific differentiation of the immortalized BEpsilon in vitro. Specific Aim 1: Resolution of the tolerogenic IgE epitope: To stepwise (i) examine the molecular conservation of this epitope among different IgE molecules, (ii) localize the tolerogenic domain by enzymatic cleavage, and constructing this epitope through synthetic peptide techniques. Specific Aim 2: Construction of immortal BEpsilon-cell lines: This aim is to be achieved by applying our expertise in the growth of B-cell lines utilizing B-cell growth factors (BCGF) and our newly developed DNA transfection technique for inserting transforming genes into lymphoid cells. The primary BEpsilon-cell candidates for transfection are from IgE-defective SJA/20 and normal BALB/c mice. Specific Aim 3: Elucidation of control mechanism of clonal negative regulatory T-cells on the stage-specific differentiation of BEpsilon-cell lines. To demonstrate a forward differentiation of BEpsilon-cells at different maturation stages and to modulate these processes by regulatory T-cells and factors prepared from IgE tolerant mice. Subsequently clonal class-specific regulatory T-cells are examined for their direct or indirect effects on BEpsilon-cells. Contributions from this project will be made to two areas. First, an in vitro IgE antibody system will serve as a paradigm for class-specific regulation of other isotypes. Second, the identification of the minimum tolerogenic epitope in IgE will enable the preparation of a safe and efficient vaccine for IgE-mediated disorders provoked by diverse allergens.

免疫球蛋白E（IgE）类抗体的产生与 人类和实验动物的过敏性疾病。 我们 对这类免疫的理解在1990年代得到了极大的扩展， 通过对IgE结构和功能的阐明， 分子。 最近，我们发现了一种新的模型系统， 在用IgE配体体外处理的小鼠中的多克隆IgE耐受性。 然而，关于调节细胞如何发挥作用， 控制EgE的B细胞的阶段特异性分化 谱系（BEpsilon-细胞）。 这种特殊的困难主要是由于 缺乏可再现体外IgE抗体系统。 的目的 研究建议是确定固有的精细致耐受性表位， IgE分子，并检查类特异性阴性细胞如何影响 体外永生化的BEp细胞的阶段特异性分化。 具体目的1：耐受原性IgE表位的解析： 为了逐步（i）检查该表位的分子保守性， 不同的IgE分子，（ii）通过酶促免疫反应定位致耐受性结构域， 切割，并通过合成肽构建该表位， 技术. 具体目的2：构建永生BEpsilon细胞系： 这一目标将通过将我们的专业知识应用于以下方面来实现： 利用B细胞生长因子（BCGF）的B细胞系和我们新开发的 将转化基因插入淋巴细胞的DNA转染技术 细胞 用于转染的原代BEpsilon细胞候选物来自 IgE缺陷型SJA/20和正常BALB/c小鼠。 具体目的3：阐明克隆阴性的控制机制 调节性T细胞对BEp细胞阶段特异性分化的影响 线 为了证明在不同的条件下BEpsilon细胞的正向分化， 成熟阶段，并通过调节性T细胞调节这些过程， 从IgE耐受小鼠制备的因子。 后无性系 检查类别特异性调节性T细胞的直接或间接 对BEpsilon细胞的影响。 该项目将对两个领域作出贡献。 首先，一个 体外IgE抗体系统将作为类特异性的范例 其他同种型的调节。 二、最低限度的认定 IgE中的致耐受性表位将使得能够制备安全且 针对由多种过敏原引起的IgE介导的疾病的有效疫苗。