McKeever P E, Garvin A J, Hardin D H, Spicer S S
Am J Pathol. 1976 Sep;84(3):437-56.
A recently developed method for ultrastructural demonstration of cell surface receptors for immune complexes is applied to evaluation of these receptors on various cell types. The method entailing incubation with a complex of horesradish peroxidase (HRP) and antibody to HRP (anti-HRP) disclosed dense foci indicative of immune complex receptors distributed at 30- to 120-mmu intervals over macrophage surfaces. Invaginations, loop-like evaginations, and pinocytotic vasicles stained prominently. The number of stained immune complex receptors averaged 200,000 per oil-induced macrophage and 120,000 per noninduced macrophage, as determined from counts of focal deposits in electron micrographs. Receptor periodicity on giant cells present in oil-induced exudates resembled that on macrophages, but the larger giant cells contained an estimated 1.5 million sites. Although receptor periodicity on eosinophils and neutrophils equaled that on macrophages, the staining was lighter and was interrupted by intervals of unstained membrane. Neutrophils averaged 28,000 and eosinophils 35,000 receptors per cell, whereas those lymphocytes with receptors averaged 3,500 per cell. Viable cells incubated with anti-HRP sequentially exhibited about half as many reactive sites as did cells incubated with immune complex. When warmed to 37 C, viable macrophages and eosinophils pinocytosed soluble immune complexes almost completely within 30 minutes and phagocytosed insoluble complexes more slowly. The endocytosed soluble immune complexes were sequestered within tubulovesicular structures in addition to the expected phagocytic vacuoles. Receptors appeared fully active on macrophages that were restained with soluble, cold immune complex after they had endocytosed immune complex in the course of a 30-minute warming interval.
一种最近开发的用于超微结构显示免疫复合物细胞表面受体的方法被应用于评估各种细胞类型上的这些受体。该方法包括用辣根过氧化物酶(HRP)和抗HRP抗体(抗HRP)的复合物孵育,结果显示在巨噬细胞表面有密集的病灶,表明免疫复合物受体以30至120微米的间隔分布。内陷、环状外翻和吞噬泡染色明显。根据电子显微镜照片中病灶沉积物的计数,每只油诱导的巨噬细胞上染色的免疫复合物受体平均数量为200,000个,每只未诱导的巨噬细胞为120,000个。油诱导渗出物中存在的巨细胞上的受体周期性与巨噬细胞上的相似,但较大的巨细胞估计含有150万个位点。尽管嗜酸性粒细胞和中性粒细胞上的受体周期性与巨噬细胞上的相等,但染色较浅且被未染色的膜间隔中断。中性粒细胞平均每个细胞有28,000个受体,嗜酸性粒细胞平均每个细胞有35,000个受体,而有受体的淋巴细胞平均每个细胞有3,500个受体。与抗HRP顺序孵育的活细胞显示的反应位点数量约为与免疫复合物孵育细胞的一半。当加热到37℃时,活的巨噬细胞和嗜酸性粒细胞在30分钟内几乎完全吞噬可溶性免疫复合物,吞噬不溶性复合物的速度较慢。除了预期的吞噬泡外,内吞的可溶性免疫复合物还被隔离在管状小泡结构内。在30分钟的升温间隔内巨噬细胞内吞免疫复合物后,用可溶性冷免疫复合物重新染色时,受体似乎完全活跃。