Kubota T, Handa Y, Tsuchida A, Kaneko M, Kobayashi H, Kubota T
Department of Neurosurgery, Fukui Medical School, Japan.
Stroke. 1993 Dec;24(12):1993-2000; discussion 2000-1. doi: 10.1161/01.str.24.12.1993.
Although it has been suggested that humoral immunity plays a role in the pathogenesis of cerebral vasospasm after subarachnoid hemorrhage, there has been no quantitative assay for cellular immunity. We studied the kinetics of immune cells in the subarachnoid space after subarachnoid hemorrhage in the rat.
One hundred fourteen Sprague-Dawley rats were used in this study. The animals were divided into two groups and injected with either autologous blood (0.3 mL) or saline into the major cistern. They were killed at the specified time: 10 minutes or 1, 2, 3, 5, or 7 days after subarachnoid hemorrhage. For immunohistochemical analysis, the rats' whole brains were frozen, and cryostat sections were prepared. For flow cytometric analysis of immune cell presence, their whole brains underwent enzymatic digestion.
Histopathologic study revealed pathological change of the arterial wall, and immunohistochemical study revealed the existence of macrophages and T cells in the subarachnoid space in animals with a survival time of 2 to 5 days after subarachnoid hemorrhage. A flow cytometric study revealed the peak of appearance of T cells and macrophages 2 days after subarachnoid hemorrhage. The helper-suppressor T cell ratio also reached a peak 2 days after subarachnoid hemorrhage.
A serial response of immunoreactive cells, which resembles that of the chronic allergic reaction observed in autoimmune diseases or delayed-type hypersensitivity, exists in the subarachnoid space after subarachnoid hemorrhage. The present results suggest that the initial response in cellular immunity, which is followed by humoral immunity and eicosanoid reactions, plays a role in eliciting the development of cerebral vasospasm.
尽管有研究表明体液免疫在蛛网膜下腔出血后脑血管痉挛的发病机制中起作用,但尚无针对细胞免疫的定量检测方法。我们研究了大鼠蛛网膜下腔出血后蛛网膜下腔内免疫细胞的动力学变化。
本研究使用了114只Sprague-Dawley大鼠。将动物分为两组,分别向大脑大池注射自体血(0.3 mL)或生理盐水。在蛛网膜下腔出血后的特定时间点(10分钟或1、2、3、5或7天)处死动物。用于免疫组织化学分析时,将大鼠全脑冷冻,制备冰冻切片。用于免疫细胞存在情况的流式细胞术分析时,对全脑进行酶消化。
组织病理学研究显示动脉壁有病理改变,免疫组织化学研究显示蛛网膜下腔出血后存活2至5天的动物中存在巨噬细胞和T细胞。流式细胞术研究显示蛛网膜下腔出血后2天T细胞和巨噬细胞出现的峰值。辅助性T细胞与抑制性T细胞的比例在蛛网膜下腔出血后2天也达到峰值。
蛛网膜下腔出血后蛛网膜下腔内存在免疫反应性细胞的系列反应,类似于自身免疫性疾病或迟发型超敏反应中观察到的慢性过敏反应。目前的结果表明,细胞免疫的初始反应继之以体液免疫和类花生酸反应,在引发脑血管痉挛的发展中起作用。