Räsänen L, Lehto M, Leinikki P
J Immunol Methods. 1984 Dec 31;75(2):361-8. doi: 10.1016/0022-1759(84)90120-0.
Lymphocytes bind certain bacteria. This property has been utilized in the lymphocyte bacterial rosette assay to identify T and B cell subsets. Here we determined the optimum conditions for the assay, studied the effect of metabolic inhibitors and divalent cations and compared bacterial rosette-forming lymphocyte subpopulations with those defined with monoclonal antibodies. The strains used were Brucella melitensis, Bacillus subtilis, Staphylococcus aureus and Escherichia coli. Optimum attachment was obtained at 4 degrees C in 6% BSA with ultrasonicated bacteria. Pretreatment of lymphocytes with the microfilament-disruptive drug cytochalasin B suppressed the binding of bacteria, whereas colchicine (inhibitor of microtubules), puromycin (inhibitor of translation), sodium azide (inhibitor of oxidative phosphorylation) and 2-deoxyglucose (inhibitor of glycolysis) had no effect. Divalent cations were required for the attachment of bacteria. B. melitensis bound to DR-positive cells, whereas the other bacterial strains rosetted OKT4- OKT8- and DR-positive cells without exhibiting helper or suppressor T cell or B lymphocyte specificity.
淋巴细胞可结合某些细菌。这一特性已被用于淋巴细胞细菌花环试验以鉴定T细胞和B细胞亚群。在此，我们确定了该试验的最佳条件，研究了代谢抑制剂和二价阳离子的作用，并将形成细菌花环的淋巴细胞亚群与用单克隆抗体定义的亚群进行了比较。所使用的菌株为羊布鲁氏菌、枯草芽孢杆菌、金黄色葡萄球菌和大肠杆菌。在4℃、6%牛血清白蛋白（BSA）中使用超声处理过的细菌可获得最佳结合效果。用微丝破坏药物细胞松弛素B预处理淋巴细胞可抑制细菌的结合，而秋水仙碱（微管抑制剂）、嘌呤霉素（翻译抑制剂）、叠氮化钠（氧化磷酸化抑制剂）和2-脱氧葡萄糖（糖酵解抑制剂）则无此作用。细菌的结合需要二价阳离子。羊布鲁氏菌与DR阳性细胞结合，而其他细菌菌株则与OKT4 - OKT8 -和DR阳性细胞形成花环，且未表现出辅助性或抑制性T细胞或B淋巴细胞特异性。
