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

与抗原提呈细胞的接触启动了一种激活 T淋巴细胞内的级联反应，包括细胞内钙的升高， 淋巴因子的产生和细胞分裂。结合钙离子成像 启用光学陷阱后，T细胞接触要求和 极性将在单细胞水平上进行研究。抗CD3抗体 单抗包裹的微球诱导的钙信号高出约10倍 与T细胞前沿接触时的频率比较 有后缘的。至少340个T细胞受体的结合 (细胞上总钙的~1%)需要启动钙离子 信号传递，最小接触面积为~3(m2.我们的结果 表示需要约1000个TCR才能生成持久的 [Ca~(2+)]I信号，这可能是基因表达所必需的。我们预计 这种方法可以与辅助分子的配体结合，或者 与下游的基因表达分析，以检查物种 T细胞中的克隆特异性变异反应并加深了我们的 对早期激活事件与脑电活动关系的认识 基因表达。

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.