Laboratory of Nanophotonic Functional Materials and Devices, South China Normal University, Guangzhou, Guangdong 510006, China.
Biomed Res Int. 2013;2013:276498. doi: 10.1155/2013/276498. Epub 2013 Oct 30.
Direct molecular imaging of nanoscale relationship between T-cell receptor complexes (TCR/CD3) and gangliosidosis GM1 before and after T-cell activation has not been reported. In this study, we made use of our expertise of near-field scanning optical microscopy(NSOM)/immune-labeling quantum dots- (QD-)based dual-color imaging system to visualize nanoscale profiles for distribution and organization of TCR/CD3, GM1, as well as their nanospatial relationship and their correlation with PKC θ signaling cascade during T-cell activation. Interestingly, after anti-CD3/anti-CD28 Ab co-stimulation, both TCR/CD3 and GM1 were clustered to form nanodomains; moreover, all of TCR/CD3 nanodomains were colocalized with GM1 nanodomains, indicating that the formation of GM1 nanodomains was greatly correlated with TCR/CD3 mediated signaling. Specially, while T-cells were pretreated with PKC θ signaling inhibitor rottlerin to suppress IL-2 cytokine production, no visible TCR/CD3 nanodomains appeared while a lot of GM1 nanodomains were still observed. However, while T-cells are pretreated with PKCα β signaling inhibitor GÖ6976 to suppress calcium-dependent manner, all of TCR/CD3 nanodomains were still colocalized with GM1 nanodomains. These findings possibly support the notion that the formation of GM1 nanodomains indeed serves as platforms for the recruitment of TCR/CD3 nanodomains, and TCR/CD3 nanodomains are required for PKCθ signaling cascades and T-cell activation.
尚未有报道直接对 T 细胞受体复合物（TCR/CD3）与神经节苷脂贮积症 GM1 之间的纳米级关系在 T 细胞激活前后进行分子成像。在这项研究中，我们利用近场扫描光学显微镜（NSOM）/免疫标记量子点（QD）双色彩像系统的专业知识，可视化 TCR/CD3、GM1 的分布和组织的纳米级轮廓，以及它们在 T 细胞激活过程中的纳米级空间关系及其与 PKCθ信号级联的相关性。有趣的是，在用抗-CD3/抗-CD28 Ab 共刺激后，TCR/CD3 和 GM1 都聚集形成纳米域；此外，所有 TCR/CD3 纳米域都与 GM1 纳米域共定位，表明 GM1 纳米域的形成与 TCR/CD3 介导的信号传递密切相关。特别地，当 T 细胞用 PKCθ信号抑制剂rottlerin 预处理以抑制 IL-2 细胞因子的产生时，虽然没有观察到可见的 TCR/CD3 纳米域，但仍观察到大量 GM1 纳米域。然而，当 T 细胞用 PKCαβ信号抑制剂 GÖ6976 预处理以抑制钙依赖性方式时，所有 TCR/CD3 纳米域仍与 GM1 纳米域共定位。这些发现可能支持这样一种观点，即 GM1 纳米域的形成确实充当了 TCR/CD3 纳米域募集的平台，并且 TCR/CD3 纳米域是 PKCθ信号级联和 T 细胞激活所必需的。
