Hummel J L, Lichty B D, Reis M, Dubé I, Kamel-Reid S
Institute of Medical Sciences, Graduate Department of Cellular and Molecular Pathology, University of Toronto, Ontario, Canada.
Leukemia. 1996 Aug;10(8):1370-6.
Chronic lymphocytic leukemia (CLL) is a heterogeneous disease of the elderly which can present in one of three stages; benign, intermediate or advanced. The molecular events governing the progression of CLL are poorly understood. In order to develop model systems for predicting the aggressiveness of leukemic clones in CLL, in vivo transplantation of SCID mice with CLL cells, and the in vitro growth of CLL cells on mouse and human stromal layers, were investigated. Bone marrow or peripheral blood cells from 40 patients at different stages of CLL were transplanted into 172 immune-deficient SCID mice. Thirty-five percent of SCID mice injected with CLL cells were positive for the presence of human DNA by Southern blot or PCR analysis. The most frequently involved sites were the spleen, lung, kidney and bone marrow, at levels corresponding from 0.1 to 10 percent human DNA. Thrice-weekly intraperitoneal injections of IL-2, alone or in combination with IL-7, did not increase the level of human cell engraftment. SCID mice developed endogenous thymic lymphomas at an incidence of 10-33 percent, a rate that was not increased by CLL cell transplantation. In vitro, CLL cells were able to proliferate for 9 weeks on human stromal layers supplemented with CM (conditioned media from a culture of the human bladder carcinoma cell line 5637), but failed to thrive on the murine stromal cell line MTE cultured either in CM or autologous serum. FACS analysis revealed that 81 percent of proliferating cells on human stromal layers carried the CD5 cell surface marker, identifying them as CLL cells. Previously EBV-negative CLL cells became EBV-positive after 9 to 12 weeks in culture. The results of this study provide a firm foundation for the development of in vivo and in vitro model systems for the study of human CLL.
慢性淋巴细胞白血病（CLL）是一种常见于老年人的异质性疾病，可分为三个阶段之一：良性、中间型或晚期。目前对控制CLL进展的分子事件了解甚少。为了开发预测CLL中白血病克隆侵袭性的模型系统，研究了将CLL细胞移植到SCID小鼠体内以及CLL细胞在小鼠和人类基质层上的体外生长情况。将40例处于不同阶段的CLL患者的骨髓或外周血细胞移植到172只免疫缺陷的SCID小鼠体内。通过Southern印迹或PCR分析，35%注射CLL细胞的SCID小鼠检测到人类DNA阳性。最常受累的部位是脾脏、肺、肾脏和骨髓，人类DNA水平在0.1%至10%之间。每周三次腹腔注射IL-2，单独或与IL-7联合使用，均未提高人类细胞植入水平。SCID小鼠发生内源性胸腺淋巴瘤的发生率为10%-33%，CLL细胞移植并未增加该发生率。在体外，CLL细胞能够在添加了CM（来自人膀胱癌细胞系5637培养的条件培养基）的人类基质层上增殖9周，但在CM或自体血清中培养的小鼠基质细胞系MTE上无法生长。FACS分析显示，在人类基质层上增殖的细胞中有81%携带CD5细胞表面标志物，表明它们为CLL细胞。先前EBV阴性的CLL细胞在培养9至12周后变为EBV阳性。本研究结果为开发用于研究人类CLL的体内和体外模型系统奠定了坚实基础。
