Crosstalk of immune and nervous systems by 3-D imaging

很长一段时间以来，众所周知，思想可能影响身体。与此相关的是，神经和免疫系统之间的双向通信的想法已经被广泛讨论。它基于实验证据，表明神经递质和神经元活动可以影响免疫细胞并诱导免疫反应。另一方面，免疫激活会影响中枢和自主神经系统的功能。在这里，为了了解免疫和神经系统之间这种通讯的分子机制，我们与麦克马斯特大学的John Bienenstock教授合作，建立了小鼠症状神经元与大鼠嗜碱粒细胞白血病(RBL-2H3)细胞(粘膜肥大细胞的同源物)共培养的时间培养模型。在共培养3天后，光镜观察到轴突与RBL-2H3细胞接触。然后，我们用激光共聚焦扫描显微镜研究了相互作用后神经细胞和肥大细胞中的钙信号。神经突起经缓激肽刺激后，荧光强度增加。这表明刺激缓激肽后，神经突起内钙离子浓度升高。之后，与神经突起接触的RBL-2H3细胞内钙离子浓度也升高。然而，在未与突起接触的RBL-2H3细胞中从未观察到这种钙信号。用蝎毒刺激RBL-2H3细胞，也能诱导出类似的钙信号。通过使用抗P物质抗体的实验，我们发现从神经突起释放的P物质在RBL-2H3细胞中触发了这些钙信号。

It has been well known for a long time that the mind may influence the body. Related to this, the idea of bidirectional communication between nervous and immune systems has been discussed extensively. It is based on experimental evidence showing that neurotransmitters and neuronal activity can affect immune cells and induce immunological responses. On the other hand, immune activation can influence the function of the central and autonomic nervous systems. Here, to understand the molecular mechanism of these kinds of communication between the immune and nervous systems we established a time culture model of murine symphathetic neurons co-cultured with rat basophilic leukaemia (RBL-2H3) cells (homologues of mucosal mast cells) in collaboration with Prof.John Bienenstock at McMaster University. In co-cultures of up 3 days, light microscopy identified neurite contacts with RBL-2H3 cells. Then, we have studied the calcium signals in both nerve and mast cells after the interaction by confocal laser scanning microscopy. Fluo-3 fluorescence intensity in the neurites increased after stimulation with bradykinin. this indicated that calcium ion concentration increased in the neurites with bradykinin stimulation. After that calcium ion concentration increased also in RBL-2H3 cells which were contacted with neurites. However, such kinds of calcium signals were never observed in RBL-2H3 cells which were not contacted with the neurites. Similar kinds of calcium signals were also induced in RBL-2H3 cells when the neurites were stimulated with scorpion venom. By the experiments using anti-substance P antibody, we found that substance P,which were released from the neurites, triggered these calcium signals in RBL-2H3 cells.