Liu C C, Perussia B, Cohn Z A, Young J D
J Exp Med. 1986 Dec 1;164(6):2061-76. doi: 10.1084/jem.164.6.2061.
We show here that human peripheral blood NK cells contain a pore-forming protein (PFP) with an Mr of 70,000-72,000 that assembles structural lesions (with an average internal diameter of 150-170 A) and forms functional channels. The PFP was isolated by affinity chromatography from human NK cells, using a specific anti-C9 antiserum as the immunoadsorbent. The NK cells were isolated from PBL by positive or negative selection by indirect rosetting using a panel of monoclonal antibodies directed against different NK and T cell surface antigens. PFP was identified in NK cells freshly isolated and isolated from cultured PBL, both stimulated with interleukin 2, but not in NK cell-depleted lymphocytes. In planar bilayers, the channels formed by the NK cell-derived PFP are highly voltage resistant, with most channels persisting in the open state once they have inserted into the bilayer. The unit conductances of these channels range 0.3-1 nS in 0.1 M NaCl. The channels show poor selectivity for monovalent and divalent ions. The PFP is also released from human NK cells stimulated with the calcium ionophore A23187, suggesting that this protein, like the one produced by murine CTL lines, may be similarly secreted during cell-mediated killing. Its identification in primary human NK cell cultures indicates that this protein may play an active role in NK cell-mediated killing.
我们在此表明，人类外周血自然杀伤细胞（NK细胞）含有一种分子量为70,000 - 72,000的成孔蛋白（PFP），该蛋白能形成结构损伤（平均内径为150 - 170埃）并形成功能性通道。使用特异性抗C9抗血清作为免疫吸附剂，通过亲和层析从人类NK细胞中分离出PFP。通过使用一组针对不同NK和T细胞表面抗原的单克隆抗体，通过间接玫瑰花结法进行阳性或阴性选择，从外周血淋巴细胞（PBL）中分离出NK细胞。在新鲜分离的NK细胞以及从培养的PBL中分离出的、经白细胞介素2刺激的NK细胞中均鉴定出了PFP，但在去除NK细胞的淋巴细胞中未鉴定出。在平面双层膜中，NK细胞来源的PFP形成的通道具有高度电压抗性，一旦插入双层膜，大多数通道会保持开放状态。在0.1 M NaCl中，这些通道的单位电导范围为0.3 - 1 nS。这些通道对单价和二价离子的选择性较差。用钙离子载体A23187刺激人类NK细胞时，PFP也会释放出来，这表明这种蛋白与小鼠细胞毒性T淋巴细胞（CTL）系产生的蛋白类似，可能在细胞介导的杀伤过程中以类似方式分泌。在原代人类NK细胞培养物中鉴定出该蛋白，表明它可能在NK细胞介导的杀伤中发挥积极作用。
