Most acute myeloid leukemia progenitor cells with long-term proliferative ability in vitro and in vivo have the phenotype CD34+/CD71-/HLA-DR-
Most acute myeloid leukemia progenitor cells with long-term proliferative ability in vitro and in vivo have the phenotype CD34+/CD71-/HLA-DR-
复制标题
DOI：
10.1182/blood.v92.11.4325
复制
发表时间：
1998-12-01
期刊：
影响因子：
20.3
通讯作者：
Sutherland, HJ
中科院分区：
文献类型：
--
作者：
Blair, A;Hogge, DE;Sutherland, HJ
急性髓系白血病 (AML) 是造血祖细胞恶性转化的结果，造血祖细胞增殖形成 AML 母细胞积聚。这些 AML 细胞中只有少数能够在体外增殖，这表明 AML 细胞可能按层次结构组织，其中只有最原始的细胞能够维持白血病克隆。为了进一步研究这一假设，我们评估了基于表面抗原表达纯化这些原始细胞的策略。作为体外终点，我们确定了能够在悬浮培养 (SC) 中产生长达 8 周的 AML 集落形成细胞 (CFU) 的 AML 祖细胞的表型，并将该表型与在非肥胖糖尿病/严重联合免疫缺陷 (NOD/SCID) 小鼠中复制 AML 的细胞的表型进行比较。对 AML 细胞进行荧光激活细胞分选 (FACS)，以检测 CD34 和 CD71、CD38 和/或 HLA-DR 的共表达，并在体外和体内以不同的细胞剂量对亚组分进行分析，以评估纯化情况。虽然大多数原发性 AML CFU 缺乏 CD34 的表达，但大多数在 SC 中 2 至 8 周后能够产生 CFU 的细胞是 CD34(+)/CD71(-)。 2 至 4 周后，产生 CFU 的细胞中的 HLA-DR 表达是异质的。然而，在 SC 6 至 8 周后，大部分 CFU 来自 CD34(+)/HLA-DR- 细胞。同样，大多数能够从 SC 长期产生 CFU 的细胞是 CD34(+)/CD38(-)。大多数能够移植NOD/SCID小鼠的细胞也是CD34+/CD71-和CD34(+)/HLA-DR-。 CD34(+)/CD71(+)或HLA-DR+亚组分未实现植入，然而，在两名患者中，CD34(+)和CD34(-)亚组分均能够植入NOD/SCID小鼠。结合这些抗原的三色分选策略可以对这些 NOD/SCID 白血病起始细胞进行大约 2 个对数级的纯化，并且在一次实验中仅使用 400 个细胞即可实现植入。表型研究表明，通过将缺乏 CD38 表达与 CD34(+)/CD71(-) 或 CD34(+)/HLA-DR- 表型相结合，可以实现更高的纯化。这些结果表明，大多数能够在体外和体内长期增殖的AML细胞与正常干细胞共享CD34(+)/CD71(-)/HLA-DR-表型。我们的数据表明，在这组患者中，根据表型定义，白血病转化发生在原始祖细胞中，并根据功能分析定义，随后发生了某种程度的分化。 (C) 1998 年，美国血液学会。
Acute myeloid leukemia (AML) occurs as the result of malignant transformation in a hematopoietic progenitor cell, which proliferates to form an accumulation of AML blasts. Only a minority of these AML cells are capable of proliferation in vitro, suggesting that AML cells may be organized in a hierarchy, with only the most primitive of these cells capable of maintaining the leukemic clone. To further investigate this hypothesis, we have evaluated a strategy for purifying these primitive cells based on surface antigen expression. As an in vitro endpoint, we have determined the phenotype of AML progenitor cells which are capable of producing AML colony-forming cells (CFU) for up to 8 weeks in suspension culture (SC) and compared the phenotype with that of cells which reproduce AML in nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice. AML cells were fluorescence-activated cell sorted (FACS) for coexpression of CD34 and CD71, CD38, and/or HLA-DR and the subfractions were assayed in vitro and in vivo at various cell doses to estimate purification. While the majority of primary AML CFU lacked expression of CD34, most cells capable of producing CFU after 2 to 8 weeks in SC were CD34(+)/CD71(-). HLA-DR expression was heterogeneous on cells producing CFU after 2 to 4 weeks. However, after 6 to 8 weeks in SC, the majority of CFU were derived from CD34(+)/HLA-DR- cells. Similarly, the majority of cells capable of long-term CFU production from SC were CD34(+)/CD38(-). Most cells that were capable of engrafting NOD/SCID mice were also CD34+/CD71- and CD34(+)/HLA-DR-. Engraftment was not achieved with CD34(+)/CD71(+) or HLA-DR+ subfractions, however, in two patients, both the CD34(+) and CD34(-) subfractions were capable of engrafting the NOD/SCID mice. A three-color sorting strategy combining these antigens allowed approximately a 2-log purification of these NOD/SCID leukemia initiating cells, with engraftment achieved using as few as 400 cells in one experiment. Phenotyping studies suggest even higher purification could be achieved by combining lack of CD38 expression with the CD34(+)/CD71(-) or CD34(+)/HLA-DR- phenotype. These results suggest that most AML cells capable of long-term proliferation in vitro and in vivo share the CD34(+)/CD71(-)/HLA-DR- phenotype with normal stem cells. Our data suggests that in this group of patients the leukemic transformation has occurred in a primitive progenitor, as defined by phenotype, with some degree of subsequent differentiation as defined by functional assays. (C) 1998 by The American Society of Hematology.