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首页> 美国卫生研究院文献>Proceedings of the National Academy of Sciences of the United States of America >Mapping the sensitivity of T cells with an optical trap: Polarity and minimal number of receptors for Ca2+ signaling
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Mapping the sensitivity of T cells with an optical trap: Polarity and minimal number of receptors for Ca2+ signaling

机译：用光阱测绘T细胞的敏感性：极性和最少数量的Ca2 +信号转导受体

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Contact with antigen-presenting cells (APCs) initiates an activation cascade within T lymphocytes, including a rise in cytosolic calcium, lymphokine production, and cell division. Although T cell–APC physical contact is required for an immune response, little is known about the patterns of cellular interactions and their relation to activation. Calcium imaging combined with an optical trap enabled the T cell contact requirements and polarity to be investigated at the single-cell level. APCs or anti-CD3 mAb-coated beads were trapped with a laser and placed at different locations along the T cell, which has a polarized appearance defined by the shape and direction of crawling. T cells were 3-fold more sensitive to APC contact made at the leading edge of the T cell than with contact made at the tail. Anti-CD3 mAb-coated 6-μm beads induced calcium signaling with ≈10-fold higher frequency and ≈4-fold shorter latency on contact with the leading edge of the T cell than on contact with the trailing edge. Alterations in antibody density (2 to 500 per μm2) and bead size (1 to 6 μm in diameter) were used to determine the spatial requirements and the minimal number of receptors which must be engaged to transmit a positive signal. T cell response percentage, latency, and calcium-signaling pattern (transient vs. sustained or oscillatory) depended on antibody density on the bead. The presence of ≈170 anti-CD3 mAb within the contact area elicited a detectable T cell calcium response. We propose here that engagement of no more than 340 T cell receptors (≈1% of the total on the cell) is sufficient to initiate Ca2+ signaling. The minimal contact area was ≈3 μm2.
机译：与抗原呈递细胞（APC）的接触可启动T淋巴细胞内的激活级联反应，包括胞质钙的增加，淋巴因子的产生和细胞分裂。尽管免疫反应需要T细胞与APC的物理接触，但对细胞相互作用的方式及其与激活的关系知之甚少。钙成像与光阱相结合使得能够在单细胞水平上研究T细胞的接触要求和极性。用激光捕获APC或涂有抗CD3 mAb的珠子，并将其放置在沿T细胞的不同位置，T细胞的极化外观由爬行的形状和方向定义。 T细胞对在T细胞前缘进行的APC接触的敏感性比在尾部进行的接触的敏感性高3倍。用抗CD3 mAb包裹的6-μm磁珠诱导钙信号传导，与T细胞前缘接触的频率比与后缘接触的频率高约10倍，潜伏期短约4倍。抗体密度（每μm 2 为2至500）和珠子大小（直径为1至6μm）的变化被用于确定空间要求和必须结合的最少数量的受体才能传递抗体。正信号。 T细胞应答百分比，潜伏期和钙信号传导模式（瞬时与持续或振荡）取决于珠子上的抗体密度。接触区域内≈170抗CD3 mAb的存在引发了可检测的T细胞钙反应。我们在此提出，不超过340个T细胞受体的参与（约占细胞总数的1％）足以启动Ca 2 + 信号传导。最小接触面积为≈3μm 2 。

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