[阵发性夜间血红蛋白尿患者骨髓造血细胞对粒细胞集落刺激因子的反应]

[The response of bone marrow hematopoietic cells to G-CSF in paroxysmal nocturnal hemoglobinuria patients].

作者信息

Cao Yan-Ran, Shao Zong-Hong, Liu Hong, Shi Jun, Bai Jie, Tu Mei-Feng, Wang Hua-Quan, Xing Li-Min, Cui Zhen-Zhu, Sun Juan, Jia Hai-Rong, Yang Tian-Ying

机构信息

Institute of Hematology and Blood Diseases Hospital, CAMS & PUMC, Tianjin 300020, China.

出版信息

Zhonghua Xue Ye Xue Za Zhi. 2005 Apr;26(4):235-8.

PMID:15949269

Abstract

OBJECTIVE

To study the response of hematopoietic cells (HSC) to granulocyte colony stimulating factor (G-CSF) in paroxysmal nocturnal hemoglobinuria (PNH) patients.

METHODS

(1) Bone marrow mononuclear cells (BMMNC) from 17 PNH patients and 12 normal subjects were inoculated into semisolid culture media containing or not G-CSF (50 ng/ml). The cluster/colony forming unit-granulocyte/monocyte (CFU/cFU-GM) were counted and compared. (2) BMMNC of 20 PNH patients and 12 normal controls were triply stained for CD34, CD59 and G-CSF receptor CD114/stem cell factor receptor (C-KIT) CD117 and assessed by FCM. The CD34(+) cells were identified as CD34(+)/CD59(+) and CD34(+)/CD59(-). Percentage of CD114 and CD117 expression in each cell population was calculated.

RESULTS

(1) PNH cFU-GM without G-CSF were (112.41 +/- 22.74)/10(5) BMMNC, while with G-CSF: (133.82 +/- 25.85)/10(5) BMMNC and normal cFU-GM were (190.33 +/- 36.05)/10(5) BMMNC, (309.42 +/- 92.94)/10(5) BMMNC, respectively. Whether with or without G-CSF, PNH BMMNC formed less cFU-GM than control did, both of the two kinds of BMMNC responded to G-CSF well (P < 0.05), but the increment of PNH cFU-GM yields was less than that of the normal control (P < 0.05). CFU-GM yields of PNH BMMNC without G-CSF were (24.29 +/- 9.05)/10(5) BMMNC, with G-CSF were (27.53 +/- 10.65)/10(5) BMMNC, while normal control were (77.42 +/- 36.01)/10(5) BMMNC and (98.00 +/- 43.14)/10(5) BMMNC, respectively. Whether with or without G-CSF, PNH BMMNC showed less CFU-GM yields than that of control (P < 0.05). (2) The percentage of CD114 positive cells in PNH CD34(+)CD59(+) BMMNC was (73.34 +/- 29.40)% and that in PNH CD34(+)CD59(-) BMMNC and in control CD34(+)CD59(+) BMMNC were (32.70 +/- 6.89)% and (58.52 +/- 29.99)%, respectively. The percentage of CD114 expression in PNH CD34(+) CD59(-) BMMNC was less than that in the other two groups (P < 0.05). The percentages of CD117 positivities on the PNH CD34(+)CD59(+) BMMNC were (76.90 +/- 22.08)%, PNH CD34(+) CD59(-) (36.03 +/- 7.69)% and control CD34(+) CD59(+) (80.28 +/- 13.36)%, respectively (P < 0.01).

CONCLUSION

In vitro, BMMNC of normal control grow better, and respond better to G-CSF than PNH BMMNC do. PNH CD34(+)CD59(-) BMMNC express less G-CSF receptor and C-KIT than PNH CD34(+)CD59(+) and normal CD34(+)CD59(+) BMMNC do, which may be the reason that abnormal PNH clone grow worse than the normal clones do.

摘要

目的

研究阵发性睡眠性血红蛋白尿（PNH）患者造血细胞（HSC）对粒细胞集落刺激因子（G-CSF）的反应。

方法

（1）将17例PNH患者和12例正常受试者的骨髓单个核细胞（BMMNC）接种于含或不含G-CSF（50 ng/ml）的半固体培养基中。计数并比较粒系/单核系集落形成单位（CFU/cFU-GM）。（2）对20例PNH患者和12例正常对照的BMMNC进行CD34、CD59和G-CSF受体CD114/干细胞因子受体（C-KIT）CD117三重染色，并用流式细胞术（FCM）进行评估。将CD34(+)细胞鉴定为CD34(+)/CD59(+)和CD34(+)/CD59(-)。计算每个细胞群中CD114和CD117表达的百分比。

结果

（1）未加G-CSF时，PNH患者的cFU-GM为（112.41±22.74）/10⁵ BMMNC，加G-CSF时为（133.82±25.85）/10⁵ BMMNC，正常对照分别为（190.33±36.05）/10⁵ BMMNC、（309.42±92.94）/10⁵ BMMNC。无论有无G-CSF，PNH患者的BMMNC形成的cFU-GM均少于对照组，两种BMMNC对G-CSF均有良好反应（P<0.05），但PNH患者cFU-GM产量的增加幅度小于正常对照（P<0.05）。未加G-CSF时，PNH患者的BMMNC的CFU-GM产量为（24.29±9.05）/10⁵ BMMNC，加G-CSF时为（27.53±10.65）/10⁵ BMMNC，而正常对照分别为（77.42±36.01）/10⁵ BMMNC和（98.00±43.14）/10⁵ BMMNC。无论有无G-CSF，PNH患者的BMMNC的CFU-GM产量均低于对照组（P<0.05）。（2）PNH患者CD34(+)CD59(+) BMMNC中CD114阳性细胞的百分比为（73.34±29.40）%，PNH患者CD34(+)CD59(-) BMMNC和正常对照CD34(+)CD59(+) BMMNC中分别为（32.70±6.89）%和（58.52±29.99）%。PNH患者CD34(+) CD59(-) BMMNC中CD114的表达百分比低于其他两组（P<0.05）。PNH患者CD34(+)CD59(+) BMMNC中CD117阳性的百分比分别为（76.90±22.08）%，PNH患者CD34(+) CD59(-)为（36.03±7.69）%，正常对照CD34(+) CD59(+)为（80.28±13.36）%（P<0.01）。

结论

体外实验中，正常对照的BMMNC生长更好，对G-CSF的反应优于PNH患者的BMMNC。PNH患者CD34(+)CD59(-) BMMNC表达的G-CSF受体和C-KIT少于PNH患者CD34(+)CD59(+)和正常CD34(+)CD59(+) BMMNC，这可能是PNH异常克隆生长不如正常克隆的原因。