Sieber G, Bochert G, Enders B, Rühl H
Blut. 1980 Aug;41(2):81-92. doi: 10.1007/BF01039652.
The kinetics of Pokeweed mitogen-induced transformation of human B-lymphocytes into immunoglobulin-secreting cells were examined in a reverse hemolytic plaque assay using protein A-coated sheep red blood cells as indicator cells. Peak responses occurred consistently after 6-8 days of incubation. After determination of the optimal experimental conditions the RHPA was found to be a reliable tool to estimate ISC in vitro; the technique was also found to be applicable for experiments surveying the B-cell response of an individual over a period of months. The PWM-induced transformation of B cells was absolutely T-cell-dependent. Other substances known as typical T-cell mitogens were also tested for polyclonal B-cell activation and some of them showed significant responses. Further experiments have shown that co-cultivation of non-HLA identical cells did not increase the number of plaques in unstimulated cultures whereas the addition of PWM leads to the generation of ISC within the expected range. These findings open a wide field of application of the RHPA in experimental and clinical immunology.
在反向溶血空斑试验中，使用蛋白A包被的绵羊红细胞作为指示细胞，检测了商陆有丝分裂原诱导人B淋巴细胞转化为免疫球蛋白分泌细胞的动力学。孵育6 - 8天后始终出现峰值反应。在确定最佳实验条件后，发现反向溶血空斑试验是体外评估免疫球蛋白分泌细胞的可靠工具；该技术也适用于长达数月的个体B细胞反应监测实验。商陆有丝分裂原诱导的B细胞转化绝对依赖于T细胞。还测试了其他已知的典型T细胞有丝分裂原对多克隆B细胞的激活作用，其中一些显示出显著反应。进一步的实验表明，非HLA匹配细胞的共培养不会增加未刺激培养物中的空斑数量，而添加商陆有丝分裂原会导致在预期范围内产生免疫球蛋白分泌细胞。这些发现为反向溶血空斑试验在实验和临床免疫学中的广泛应用开辟了广阔领域。
