Kirov S M, Kwant W O, Fernandez L A, MacSween J M, Langley G R
Br J Haematol. 1980 Feb;44(2):235-42. doi: 10.1111/j.1365-2141.1980.tb01205.x.
Chronic lymphocytic leukaemia peripheral blood mononuclear cells (CLL-PBMN) were separated into B, T and Null-enriched lymphocyte sub-populations using sequential mouse and sheep red blood cell rosetting depletions on Hypaque-Ficoll gradients. The procedure produced viable cell populations with mean percentage purities of 90, 87 and 75 for B, T and non-rosetting (Null-enriched) sub-populations, respectively. More than 80% of PBMN cells were generally accounted for by mouse and sheep rosetting. The purified lymphocyte sub-populations were examined with a panel of B-cell specific alloantisera obtained from kidney transplant recipients and a rabbit antiserum to B cell antigen isolated from a human B-lymphoblastoid line. The results illustrated that the antigens detected by these sera also have potential as a marker for characterizing the CLL population. Where conventional markers were weak or absent, B cell antigens were readily detected in both fluorescent and cytotoxic tests. The majority of the non-rosetting cells (less than 90%) in CLL followed similar patterns of reactivity to the purified B cells, suggesting they are a subset of B cells. A small residual population (0--5% of PBMN) did not react with the antisera, the significance of which is unknown.
利用在Hypaque-Ficoll梯度上依次进行小鼠和绵羊红细胞花环去除法,将慢性淋巴细胞白血病外周血单个核细胞(CLL-PBMN)分离为富含B、T和Null的淋巴细胞亚群。该方法产生的活细胞群体中,B、T和非花环形成(富含Null)亚群的平均纯度分别为90%、87%和75%。通常,超过80%的外周血单个核细胞可通过小鼠和绵羊花环形成来计数。用一组从肾移植受者获得的B细胞特异性同种抗血清以及一种从人B淋巴母细胞系分离的B细胞抗原兔抗血清,对纯化的淋巴细胞亚群进行检测。结果表明,这些血清检测到的抗原也有可能作为表征CLL群体的标志物。在传统标志物微弱或缺乏的情况下,在荧光和细胞毒性试验中均能轻易检测到B细胞抗原。CLL中大多数非花环形成细胞(不到90%)对纯化B细胞的反应模式相似,表明它们是B细胞的一个亚群。一小部分残留细胞(占外周血单个核细胞的0 - 5%)不与抗血清反应,其意义尚不清楚。