T CELL ANTIGEN RECOGNIT DYNAMIC PROBED W/ OPTICAL METHOD: CALCIUM & GENE EXPRESS

T 细胞抗原识别动态探针光学方法：钙

• 批准号: 6220406
• 负责人: MICHAEL D CAHALAN
• 金额: $ 3.09万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-01 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6220406
• 关键词: Mammalia; biomedical resource; cardiovascular system; lasers; model design /development; neoplasm /cancer; technology /technique

Contact with antigen-presenting cells initiates an activation cascade within T lymphocytes, including a rise in cytosolic calcium, lymphokine production, and cell division. Calcium imaging combined with an optical trap enabled the T-cell contact requirements and polarity to be investigated at the single-cell level. Anti-CD3 mAb-coated beads induced calcium signaling with ~ ten-fold higher frequency upon contact with the leading edge of the T cell, compared with the trailing edge. Engagement of at least 340 T cell receptors (~1% of the total on the cell) was required to initiate Ca2+ signaling, and the minimal contact area was ~3 (m2. Our results indicate that ~1000 TCRs are required to generate a long-lasting [Ca2+]i signal which might be required for gene expression. We expect this approach can be combined with ligands for accessory molecules or with downstream assays for gene expression in order to examine species and clonal-specific variations in the T cell resp onse and deepen our understanding of the relationship between early activation events and gene expression.

与抗原呈递细胞接触会激活 T淋巴细胞内的级联反应，包括胞质钙的升高， 淋巴因子产生和细胞分裂。 钙成像组合 使用光学陷阱启用了T细胞联系人要求和 极性将在单细胞水平上进行研究。 抗CD3 mab涂层珠诱导钙信号传导，高约十倍 与T细胞前缘接触时的频率，比较 带有后端。 参与至少340个T细胞受体 （启动CA2+的总数的约1％） 信号传导，最小接触面积约为3（m2。我们的结果 指出生成持久的〜1000 TCR需要 [Ca2+] I信号可能是基因表达所需的。 我们期望 该方法可以与配件分子的配体组合或 带有基因表达的下游测定法，以检查物种 和T细胞中克隆特异性的变化，并加深我们 了解早期激活事件与 基因表达。