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

与抗原呈递细胞接触会启动激活 T淋巴细胞内的级联反应，包括胞浆钙的升高， 淋巴因子的产生和细胞分裂。 钙显像联合 利用光阱实现T细胞接触要求， 在单细胞水平上研究极性。 anti-CD3 单克隆抗体包被的珠粒诱导钙信号传导， 与T细胞的前缘接触时的频率，比较 有后缘。 至少340个T细胞受体的结合 （细胞上总量的约1%）需要启动Ca 2 + 信号传导，并且最小接触面积为~3（m2. 我们的结果 这表明需要约1000个TCR来产生持久的 [Ca2+]i信号，其可能是基因表达所需的。 我们预计 该方法可以与辅助分子的配体结合 通过下游基因表达检测， 和T细胞反应中的克隆特异性变异，并加深了我们的研究。 了解早期激活事件与 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.