Kluin-Nelemans H C, Limpens J, Meerabux J, Beverstock G C, Jansen J H, de Jong D, Kluin P M
Department of Hematology, University Medical Center, Leiden, The Netherlands.
Leukemia. 1991 Mar;5(3):221-4.
A spontaneously growing EBV-negative B-cell line (DoHH2) was established from the pleural fluid cells of a 60-year-old man with centroblastic/centrocytic non-Hodgkin's lymphoma, that had transformed into an immunoblastic lymphoma. The pleural fluid cells and the DoHH2 cells expressed IgG lambda, were reactive with CD10 and CD19 monoclonal antibodies, and showed by cytogenetic analysis 48,XY, +7, +del(12)(q24), t(14;18)(q32;q21). Southern blot analysis of mini-satellite DNA patterns, and of rearrangements of the immunoglobulin genes and bcl-2, confirmed that the cell line was derived from the patient's clonal lymphoma cells. Direct nucleotide sequence analysis on polymerase chain reaction (PCR) products of the t(14;18) junction revealed an identical sequence for the JH-bcl-2 junction at JH6 and in the major breakpoint region of bcl-2 in both the original tumor cells and the DoHH2 cell line. The cell line was valuable as a standard quantification control for PCR analysis of the t(14;18) breakpoint. Titration experiments demonstrated the detection of up to one tumor cell in 10(5) normal blood lymphocytes.
一株自发生长的EBV阴性B细胞系（DoHH2）是从一名60岁患有中心母细胞/中心细胞性非霍奇金淋巴瘤且已转化为免疫母细胞性淋巴瘤的男性患者的胸水细胞中建立的。胸水细胞和DoHH2细胞表达IgG λ，与CD10和CD19单克隆抗体反应，并通过细胞遗传学分析显示为48,XY, +7, +del(12)(q24), t(14;18)(q32;q21)。对小卫星DNA模式、免疫球蛋白基因重排和bcl-2重排的Southern印迹分析证实该细胞系源自患者的克隆性淋巴瘤细胞。对t(14;18)连接处聚合酶链反应（PCR）产物的直接核苷酸序列分析显示，在原始肿瘤细胞和DoHH2细胞系中，JH6处的JH-bcl-2连接处以及bcl-2主要断裂点区域的序列相同。该细胞系作为t(14;18)断裂点PCR分析的标准定量对照很有价值。滴定实验表明，在10(5)个正常血液淋巴细胞中可检测到多达一个肿瘤细胞。
