Crosstalk of immune and nervous systems by 3-D imaging

很久以来，人们都知道，思想可以影响身体。与此相关，神经系统和免疫系统之间双向通信的想法已经被广泛讨论。它是基于实验证据表明，神经递质和神经元活动可以影响免疫细胞和诱导免疫反应。另一方面，免疫激活可以影响中枢和自主神经系统的功能。在这里，为了理解免疫系统和神经系统之间这些类型的通信的分子机制，我们与McMaster大学的John Bienenstock教授合作，建立了与大鼠嗜碱性白血病（RBL-2 H3）细胞（粘膜肥大细胞的同源物）共培养的鼠交感神经元的时间培养模型。在长达3天的共培养中，光学显微镜鉴定出神经突与RBL-2 H3细胞的接触。然后，我们用共聚焦激光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。用缓激肽刺激后，突起中的Fluo-3荧光强度增加。这表明缓激肽刺激神经突中的钙离子浓度增加。之后，在与神经突接触的RBL-2 H3细胞中钙离子浓度也增加。然而，在未与神经突接触的RBL-2 H3细胞中从未观察到这种钙信号。当用蝎毒刺激神经突起时，RBL-2 H3细胞中也诱导了类似的钙信号。通过抗P物质抗体的实验，我们发现从突起释放的P物质触发了RBL-2 H3细胞中的这些钙信号。

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.