Doubleday C W, Geoghegan W D, Jordon R E
Cutaneous Immunopathology Unit, University of Texas Health Science Center, Houston 77030.
J Lab Clin Med. 1988 Jan;111(1):28-34.
It has been reportedly previously that complement enhances pemphigus vulgaris antibody-induced epidermal cell detachment. The present studies were designed to eliminate the possibility of the human plasminogen-plasmin system contributing to the cell detachment observed in previous complement-mediated studies. To assay for the presence of protease contamination in the reagents, a simple, sensitive, caseinolytic-fluorometric assay was used. Confluent murine or human epidermal cell cultures were incubated with pemphigus IgG with and without complement and plasminogen. Detached epidermal cells, as determined by light microscopy and cell-counter analysis, were enumerated at 48 hours. Incubating pemphigus IgG with complement resulted in marked cell detachment, compared with minimal cell detachment when pemphigus IgG was incubated with plasminogen. Culture media were collected after 48 hours and assayed for activation of plasminogen. It was determined that the added plasminogen had not been activated, particularly in experiments where significant detachment was observed. The plasminogen could still be activated by streptokinase and urokinase. These studies suggest that epidermal cell detachment in pemphigus appears to be mediated at least in part by complement activation.
据先前报道,补体可增强寻常型天疱疮抗体诱导的表皮细胞脱离。本研究旨在排除人纤溶酶原 - 纤溶酶系统导致先前补体介导研究中观察到的细胞脱离的可能性。为检测试剂中蛋白酶污染的存在,使用了一种简单、灵敏的酪蛋白分解荧光测定法。将汇合的小鼠或人表皮细胞培养物与含有和不含有补体及纤溶酶原的天疱疮IgG一起孵育。通过光学显微镜和细胞计数器分析确定,在48小时时对脱离的表皮细胞进行计数。与天疱疮IgG与纤溶酶原孵育时极少的细胞脱离相比,天疱疮IgG与补体孵育导致明显的细胞脱离。48小时后收集培养基并检测纤溶酶原的激活情况。确定添加的纤溶酶原未被激活,特别是在观察到明显脱离的实验中。纤溶酶原仍可被链激酶和尿激酶激活。这些研究表明,天疱疮中的表皮细胞脱离似乎至少部分由补体激活介导。