Verstegen M M, Cornelissen J J, Terpstra W, Wagemaker G, Wognum A W
Institute of Hematology, Erasmus University Rotterdam, The Netherlands.
Leukemia. 1999 Apr;13(4):618-28. doi: 10.1038/sj.leu.2401366.
In this study the ability of malignant and normal progenitors in peripheral blood (PB) and bone marrow (BM) of CML patients in chronic phase to proliferate and produce mature progeny after transplantation into hereditary immunodeficient (SCID and NOD/SCID) mice was examined. Engraftment in NOD/SCID mice preconditioned by total body irradiation (TBI) alone was 10-fold higher than in SCID mice preconditioned by macrophage depletion and TBI, demonstrating that NOD/SCID mice are more suitable for engraftment of chronic phase CML cells. Low-density cells at cell doses of 10-30 x 10(6) and purified CD34+ cells at doses of approximately 0.2 x 10(6) engrafted NOD/SCID mice, with levels of 2 to 20% CD45+ cells with production of monocytes, granulocytes, erythroid cells, B-lymphocytes, CD34+ cells and variable frequencies of erythroid and myeloid colony-forming cells. As demonstrated by fluorescent in situ hybridization (FISH) analysis, purified human myeloid, B-lymphoid, erythroid and CD34+ cells from chimeric mouse BM contained Philadelphia-chromosome (Ph)-positive cells and Ph- cells in similar frequencies as primary cells from the CML patients. These results demonstrate that production of mature normal as well as malignant cells of multiple lineages were supported with similar efficiency. In contrast, all human erythroid and myeloid clonogenic cells detected in the mice were Ph-, which can be attributed to less efficient maintenance or more rapid differentiation of immature Ph+ cells in the mouse microenvironment. CML blast crisis cells also grew well in NOD/SCID mice, with 80-90% of human cells produced containing the Ph- chromosome. The availability of an in vivo assay that supports outgrowth of normal and malignant stem cells from chronic phase and blast crisis CML patients will facilitate examination of differential effects of growth factors, inhibitory cytokines and cytotoxic drugs on survival of normal and malignant stem cells in vivo and on progression of chronic phase CML towards blast crisis.
在本研究中，检测了慢性期慢性髓性白血病（CML）患者外周血（PB）和骨髓（BM）中的恶性祖细胞和正常祖细胞移植到遗传性免疫缺陷（SCID和NOD/SCID）小鼠后增殖并产生成熟子代的能力。仅通过全身照射（TBI）预处理的NOD/SCID小鼠中的植入率比通过巨噬细胞清除和TBI预处理的SCID小鼠高10倍，表明NOD/SCID小鼠更适合慢性期CML细胞的植入。细胞剂量为10 - 30×10⁶的低密度细胞和剂量约为0.2×10⁶的纯化CD34⁺细胞植入了NOD/SCID小鼠，CD45⁺细胞水平为2%至20%，产生单核细胞、粒细胞、红细胞、B淋巴细胞、CD34⁺细胞以及不同频率的红细胞和髓系集落形成细胞。荧光原位杂交（FISH）分析表明，来自嵌合小鼠BM的纯化人髓系、B淋巴细胞系、红细胞系和CD34⁺细胞中含有费城染色体（Ph）阳性细胞和Ph阴性细胞，其频率与CML患者的原代细胞相似。这些结果表明，多种谱系的成熟正常细胞和恶性细胞的产生得到了相似效率的支持。相比之下，在小鼠中检测到的所有人红细胞系和髓系克隆形成细胞均为Ph阴性，这可归因于在小鼠微环境中未成熟Ph⁺细胞的维持效率较低或分化更快。CML急变期细胞在NOD/SCID小鼠中也生长良好，所产生的人细胞中有80% - 90%含有Ph阴性染色体。一种支持慢性期和急变期CML患者正常和恶性干细胞生长的体内检测方法的可用性将有助于研究生长因子、抑制性细胞因子和细胞毒性药物对体内正常和恶性干细胞存活以及慢性期CML向急变期进展的不同影响。
