Xiao Juan, Wu Yong-ji, Zhang Zhi-nan, Lu Zhao-jiang, Chen Shi-ping, Dong Hong-yan
Department of Hematology, Peking Union Medical College Hospital, CAMS and PUMC, Beijing 100730, China.
Zhonghua Xue Ye Xue Za Zhi. 2003 Apr;24(4):169-73.
To explore the characteristics of CD(34)(+) CD(59)(+) cells from paroxysmal nocturnal hemoglobinuria(PNH) patients' bone marrow and the possible reasons of hematopoietic clonal dominance of PNH clones.
CD(34)(+) CD(59)(+) and CD(34)(+) CD(59)(-) cells from PNH patients and CD(34)(+) cells from normal control were selected from the bone marrow mononuclear cells by means of immunomagnetic microbead-flow cytometry two step sorting method undergone ex vivo expansion in liquid culture for two weeks and performed semisolid cultures before and after expansion.
(1) Cultivation for seven days was the optimum for ex vivo expansion of PNH CD(34)(+) CD(59)(+) cells and normal CD(34)(+) cells, both cell populations remained CD(59) positive after expansion. (2) Normal CD(34)(+) cells had higher capacities of proliferation and expansion, and stronger potential to survival than that of both PNH CD(34)(+) CD(59)(+) and PHN CD(34)(+) CD(59)(-) cells. (3) In terms of semisolid culture, there was no significant difference in the yields of CFU formation between CD(34)(+) CD(59)(+) and CD(34)(+) CD(59)(-) cells. (4) In liquid culture with combinations of hematopoietic factors SCF + IL-3 + IL-6 + FL + Tpo or SCF + IL-3 + IL-6 + FL + Tpo + Epo, there was no significant difference in the capabilities of survival, proliferation and expansion between CD(34)(+) CD(59)(+) and CD(34)(+) CD(59)(-) cells; but with combination of SCF + IL-3 + IL-6 + FL + Tpo + Epo + GM-CSF, CD(34)(+) CD(59)(-) cells had better proliferation and expansion capacities and stronger potential to survival than that of CD(34)(+) CD(59)(+) cells.
(1) Normal CD(34)(+) cells had better proliferation, expansion capacities and stronger potential to survival than that of PNH CD(34)(+) CD(59)(+)cells. (2) In semisolid and liquid culture with hematopoietic factor combinations, there was no significant difference in the capabilities of survival, proliferation and expansion between CD(34)(+) CD(59)(+) and CD(34)(+) CD(59)(-) cells. It was suggested that CD(34)(+) CD(59)(-) cells had no clonal hemotopoiesis dominance. GM-CSF might be one of the reasons for PHN clones to possess clonal hematopoiesis dominance.
探讨阵发性睡眠性血红蛋白尿症（PNH）患者骨髓中CD(34)(+) CD(59)(+)细胞的特征及PNH克隆造血克隆优势的可能原因。
采用免疫磁珠 - 流式细胞术两步分选法从骨髓单个核细胞中筛选出PNH患者的CD(34)(+) CD(59)(+)和CD(34)(+) CD(59)(-)细胞以及正常对照的CD(34)(+)细胞，在液体培养中进行体外扩增两周，并在扩增前后进行半固体培养。
（1）培养7天是PNH CD(34)(+) CD(59)(+)细胞和正常CD(34)(+)细胞体外扩增的最佳时间，扩增后两种细胞群体均保持CD(59)阳性。（2）正常CD(34)(+)细胞具有比PNH CD(34)(+) CD(59)(+)和PNH CD(34)(+) CD(59)(-)细胞更高的增殖和扩增能力以及更强的存活潜力。（3）在半固体培养方面，CD(34)(+) CD(59)(+)和CD(34)(+) CD(59)(-)细胞之间集落形成单位（CFU）形成产量无显著差异。（4）在造血因子SCF + IL - 3 + IL - 6 + FL + Tpo或SCF + IL - 3 + IL - 6 + FL + Tpo + Epo组合的液体培养中，CD(34)(+) CD(59)(+)和CD(34)(+) CD(59)(-)细胞之间的存活、增殖和扩增能力无显著差异；但在SCF + IL - 3 + IL - 6 + FL + Tpo + Epo + GM - CSF组合中，CD(34)(+) CD(59)(-)细胞比CD(34)(+) CD(59)(+)细胞具有更好的增殖和扩增能力以及更强的存活潜力。
（1）正常CD(34)(+)细胞比PNH CD(34)(+) CD(59)(+)细胞具有更好的增殖、扩增能力和更强的存活潜力。（2）在造血因子组合的半固体和液体培养中，CD(34)(+) CD(59)(+)和CD(34)(+) CD(59)(-)细胞之间的存活、增殖和扩增能力无显著差异。提示CD(34)(+) CD(59)(-)细胞无克隆造血优势。GM - CSF可能是PNH克隆具有克隆造血优势的原因之一。
