HAPTEN RECEPTOR INTERACTION IN DNAP SPECIFIC B CELL LINE

DNAP 特异性 B 细胞系中的半抗原受体相互作用

• 批准号: 3130980
• 负责人: MARLENE A. ALDO-BENSON
• 金额: $ 7.6万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-04-01 至 1988-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3130980
• 关键词: 2,4 dinitrophenol; B lymphocyte; cyclic AMP; cyclic GMP; flow cytometry; immunochemistry; leukocyte activation /transformation; lymphokines; membrane activity; phospholipids; phosphorylation; protein kinase; tissue /cell culture

The binding of antigen to receptors on B lymphocytes transmits a signal to the lymphocyte. This signal along with factors from T cells and accessory cells results in differentiation of the cell to a plasma cell secreting specific antibody. Indirect evidence suggests that the signal associated with binding of antigen and other factors is transmitted by changes in the plasma membrane and intracellular messenger. However, studies of intracellular events associated with antigen induced B cell triggering or antigen specific tolerance thus far have utilized indirect methods. Direct measurement of intracellular changes was impossible due to the small proportion of specific antigen-binding lymphocytes in most lymphocyte preparations. The principal investigator has developed long term cultures of two normal murine B cell lines which are specific to the hapten dinitrophenyl (DNP). Under the appropriate conditions, the binding of antigen (DNP-Ficoll) or tolerogen (DNP-Murine IgG) to the B cell surface receptor specific for DNP results in either antigen driven B cell differentiation or tolerance induction, respectively. This is a proposal to study the intracellular changes which occur with the binding of the hapten to the receptor on two B cell lines, and to compare the changes which occur when the antigenic and tolerance-inducing forms of the hapten are used. We intend to determine changes in intracellular levels of cylic AMP, cyclic GMP, Ca2+ and cyclic AMP-dependent protein kinases, and profiles of intracellular protein phosphorylation. We will also examine phospholipid methylation and phospholipid phosphorylation in the lymphocyte plasma membrane during induction of B cell differentiation and tolerance. Thus, we will more precisely define the biochemical events associated with hapten-receptor binding which eventually lead to B lymphocyte triggering or tolerance.

抗原与B淋巴细胞上的受体的结合将信号传递给 淋巴细胞 该信号沿着来自T细胞和辅助细胞的因子， 细胞导致细胞分化为浆细胞， 特异性抗体。 间接证据表明， 与抗原和其他因素的结合是通过改变 质膜和细胞内信使。 然而，研究 与抗原诱导的B细胞触发相关的细胞内事件，或 抗原特异性耐受性迄今为止利用间接方法。 直接 细胞内变化的测量是不可能的，因为小的 大多数淋巴细胞中特异性抗原结合淋巴细胞的比例 准备工作。 主要研究者开发了长期培养物 两个正常鼠B细胞系，其对半抗原特异 二硝基苯基（DNP）。 在适当的条件下， 抗原（DNP-Ficoll）或耐受原（DNP-Murine IgG）结合到B细胞表面 DNP特异性受体导致抗原驱动的B细胞 分化或耐受诱导。 这是一个研究细胞内变化的建议， 半抗原与两种B细胞系上的受体的结合，并比较 当抗原和耐受诱导形式的 使用半抗原。 我们打算确定细胞内 cAMP、cGMP、Ca ~（2+）和cAMP依赖蛋白水平 激酶和细胞内蛋白磷酸化的概况。 我们将 还检查了磷脂甲基化和磷脂磷酸化， 淋巴细胞质膜在B细胞分化诱导过程中 和宽容。 因此，我们将更精确地定义生物化学事件 与半抗原受体结合相关，最终导致B 淋巴细胞触发或耐受。