Moore D L, Vas S I
Immunology. 1968 Aug;15(2):185-95.
The production of complement was investigated using a modification of Jerne's haemolytic plaque technique. Guinea-pig spleen and bone marrow cells were suspended in layers of agarose containing, as indicator cells, sensitized sheep erythrocytes (EA) or intermediate products consisting of EA and some of the components of complement. R reagents or serum, in which certain components of the C′ complex had been inactivated, were added after incubation. No plaque formation due to release of complement components was observed. Bone marrow and spleen cells produced plaques when incubated with sensitized or non-sensitized erythrocytes and a reagent containing cobra venom. A method was developed for the preparation of thin films of agarose in which tissue cells and erythrocytes were suspended in a single cell layer. Single cells and clumps of cells present in the peritoneal exudate of normal guinea-pigs produced plaques when incubated with EA or EAC′ and R. Occasionally, similar clumps of cells also produced plaques with EA or EAC′ and R. The plaque-forming cells appeared to be macrophages. Plaques were not detected with R or R. No plaques were produced by spleen cells under similar experimental conditions. Bone marrow cells produced plaques with both sensitized and non-sensitized erythrocytes after prolonged incubation. Optimal plaque formation occurred when R or R was added before incubation. Plaques were not produced if the tissue cells were pre-incubated in suspension cultures for 2 hours or longer before plating. Treatment of the peritoneal exudate cells with KCN or with dinitrophenol prevented plaque formation.
采用改良的耶尔恩溶血空斑技术研究补体的产生。将豚鼠脾脏和骨髓细胞悬浮于含有致敏绵羊红细胞（EA）或由EA和部分补体成分组成的中间产物作为指示细胞的琼脂糖层中。孵育后加入已使C′复合物某些成分失活的R试剂或血清。未观察到由于补体成分释放而导致的空斑形成。当骨髓和脾脏细胞与致敏或未致敏红细胞以及含有眼镜蛇毒的试剂一起孵育时，会产生空斑。开发了一种制备琼脂糖薄膜的方法，其中组织细胞和红细胞悬浮于单细胞层中。正常豚鼠腹腔渗出液中的单个细胞和细胞团与EA或EAC′及R一起孵育时会产生空斑。偶尔，类似的细胞团与EA或EAC′及R一起也会产生空斑。形成空斑的细胞似乎是巨噬细胞。用R或R未检测到空斑。在类似实验条件下，脾脏细胞未产生空斑。长时间孵育后，骨髓细胞与致敏和未致敏红细胞均产生空斑。在孵育前加入R或R时，空斑形成最佳。如果组织细胞在平板接种前在悬浮培养中预孵育2小时或更长时间，则不会产生空斑。用KCN或二硝基苯酚处理腹腔渗出液细胞可防止空斑形成。
