Gene analysis of antibodies specific for allergen and antigenrelating to autoimmunity

过敏原和自身免疫相关抗原特异性抗体的基因分析

• 批准号: 04454070
• 负责人: KAMINOGAWA Shuichi
• 金额: $ 4.03万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04454070/
• 关键词: Immunoglobulin gene; Antibody Production; B cell; Peptide; beta-lacreglobulin; Food allevgy; MHC クラス II 分子; T細胞膜成分; IgE; クラススイッチ; ペプチド免疫; IgM産生B細胞クローン; B細胞ハイブリドーマ; gene usage; パラトープ構造

To elucidate the molecular and cellular mechanism of lgE-class switching leading to allergic diseases, we examined the differentiation process of antigen-specific B cells at a clonal level in related to the repertoire of helper T cells. We used a peptide fragment encompassing residues 25-40 (F25-40) of beta-lactoglobulin, which contained B cell determinant. T cell repertoire collaborating with F25-40-specific B cell were modified by priming with 1) beta-LG,2) F25-40-conjugated carrier protein or 3) F21-40. Thus, we demonstrate the helper functions of T cells in B cells differentiation at a clonal level by comparing primed B cell repertoire, class switching or affinity maturation among priming antigens. Consequently, the immunization with F21-40 could not induce strong lgG-class II switching. In addition, the restricted V_H gene usage (J558) was observed among B cell repertoire elicited by F21-40 distinct from other antigens. From these results, it has been suggested that T cell repertoire collaborating with B cells restrict B cell repertoire differentiating into Ab-secreting cells. To determine the molecular mechanism of the restriction, we analyzed the exact roles of the interaction between MHC class II molecules and T cell receptor on B cell activation. The results showed that the class II-mediated signals directly activate B cells and enhance lgG production. From these results, it has been suggested that the class II-mediated signals play an important role on the restriction of B cell repertoire and class switching.

为了阐明 lgE 类转换导致过敏性疾病的分子和细胞机制，我们在克隆水平上检查了与辅助 T 细胞库相关的抗原特异性 B 细胞的分化过程。我们使用了包含 β-乳球蛋白残基 25-40 (F25-40) 的肽片段，其中含有 B 细胞决定簇。通过用 1) beta-LG、2) F25-40 缀合载体蛋白或 3) F21-40 引发来修饰与 F25-40 特异性 B 细胞合作的 T 细胞库。因此，我们通过比较引发的 B 细胞库、引发抗原之间的类别转换或亲和力成熟，在克隆水平上证明了 T 细胞在 B 细胞分化中的辅助功能。因此，用F21-40免疫不能诱导强烈的IgG-II类转换。此外，在 F21-40 引发的 B 细胞库中观察到了与其他抗原不同的受限 V_H 基因使用 (J558)。这些结果表明，T 细胞库与 B 细胞协作限制了 B 细胞库分化为抗体分泌细胞。为了确定限制的分子机制，我们分析了 MHC II 类分子和 T 细胞受体之间相互作用对 B 细胞激活的确切作用。结果表明，II 类介导的信号直接激活 B 细胞并增强 IgG 的产生。这些结果表明 II 类介导的信号在 B 细胞库和类别转换的限制中发挥重要作用。

项目成果

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