Simon H B, Sheagren J N
J Exp Med. 1971 Jun 1;133(6):1377-89. doi: 10.1084/jem.133.6.1377.
An in vitro model of cellular immunity in the guinea pig was established. Animals were immunized with tubercle bacilli, bovine gamma globulin, or picrylated human serum albumin in complete Freund's adjuvant. Oil-induced peritoneal exudates from immune and control animals were cultured overnight with and without specific antigen. The cultures were washed and the macrophage monolayers were infected with Listeria monocytogenes. At intervals the monolayers were lysed and the numbers of viable intracellular bacteria were quantitated by pour plate cultures. Random monolayers were also evaluated in sequence by visually counting the intracellular bacteria on Gram-stained plates. Both methods demonstrated that the macrophages from immune animals had markedly enhanced listericidal activity when the peritoneal exudates were cultured with antigen before infection. Macrophage migration inhibition was also demonstrated under these conditions. The experiments reported here describe an in vitro model of cellular immunity which will allow separation and recombination of cell types and direct assay of cell products in efforts to elucidate further the mechanisms of the immunologically mediated enhancement of macrophage bactericidal capacity.
建立了豚鼠细胞免疫的体外模型。用结核杆菌、牛γ球蛋白或苦味酸化人血清白蛋白加完全弗氏佐剂对动物进行免疫。将免疫动物和对照动物的油诱导腹腔渗出液在有和没有特异性抗原的情况下培养过夜。培养物经洗涤后,巨噬细胞单层用单核细胞增生李斯特菌感染。每隔一段时间,裂解单层细胞,通过倾注平板培养法定量存活的细胞内细菌数量。还通过肉眼计数革兰氏染色平板上的细胞内细菌,对随机单层细胞进行顺序评估。两种方法均表明,当腹腔渗出液在感染前用抗原培养时,来自免疫动物的巨噬细胞的杀菌活性明显增强。在这些条件下还证实了巨噬细胞迁移抑制。此处报道的实验描述了一种细胞免疫的体外模型,该模型将允许细胞类型的分离和重组以及细胞产物的直接测定,以进一步阐明免疫介导的巨噬细胞杀菌能力增强的机制。