阵发性睡眠性血红蛋白尿症患者外周血淋巴细胞增殖、杀瘤活性及其对正常表型骨髓造血细胞的影响  

Proliferation and Anti-tumor activity of Lymphocytes and Their Effect on Normal Phenotype of Autologous Bone Marrow Hematopoietic Cells in Patients with Paroxysmal Nocturnal Hemoglobinuria

作　　者：王毓洲[1,2] 武永吉[1] 张之南[1] 

机构地区：[1]中国医学科学院,中国协和医科大学,北京协和医院血液科 [2]首都医科大学附属北京同仁医院呼吸科,北京100730

出　　处：《中国实验血液学杂志》2003年第1期50-53,共4页Journal of Experimental Hematology

摘　　要：阵发性睡眠性血红蛋白尿症 (PNH)克隆可能由于缺乏GPI锚连蛋白而逃脱免疫攻击 ,形成生长优势。本研究测定PNH患者淋巴细胞增殖反应及对K5 6 2细胞的杀伤作用 ,并与正常对照比较 ,观察其淋巴细胞功能。采用体外液体培养体系 ,应用免疫磁珠技术分选PNH患者骨髓CD34+ 及CD34- 细胞 ,分别向 2组细胞及对照细胞加入自身CD5 9+ 或CD5 9- 淋巴细胞及其培养上清液、外源性IFN γ及IL 2。培养 10天后 ,测定骨髓细胞中PNH细胞 (CD5 9- )百分数 ,观察淋巴细胞对骨髓中CD34+ 及CD34- 细胞中PNH细胞含量的影响。研究结果表明 ,对PHA的增殖反应 ,PNH患者未分选的淋巴细胞与健康对照比较、PNH患者自身CD5 9+ 与CD5 9- 淋巴细胞之间比较均无统计学差异 ,但对K5 6 2细胞的杀伤作用PNH患者未分选的淋巴细胞明显低于健康对照组 ,分别为 (5 0 .0 0± 2 8.6 7) %及 (76 .13± 10 .15 ) % (P <0 .0 5 ) ,而PNH患者CD5 9- 淋巴细胞与CD5 9+ 淋巴细胞之间无显著性差异。PNH患者骨髓细胞中加入自身CD5 9- 淋巴细胞或其培养上清液、CD5 9+ 淋巴细胞或其培养上清液、外源性IFN γ和IL 2培养后 ,CD5 9+ 细胞均有不同程度下降 ,除CD5 9+ 淋巴细胞组外 ,其他各组CD5 9+ 细胞下降与培养前比较有显著性差异 (P <0 .0 5 ) ,IFN γ及ILThere is a hypothesis that paroxysmal nocturnal hemoglobinuria (PNH) hematopoitic stem cells are resistant to the cytotoxic effect of T cells because they lack glycosylphosphatidylinositol (GPI) linked proteins. The aim of this study is to investigate proliferation and anti tumor activity of lymphocytes in patients with PNH, and also to assay the effect on normal cell(CD59 +) phenotype in bone marrow of PNH patient by autologous lymphocytes in vitro. MTT assay for detection of lymphocytes proliferation and its anti tumor effect was driven to delineate T cell reactive function. The CD34 - and CD34 + bone marrow cells (selected by means of immunomagnetic method) as well as unsorted marrow cells in PNH patients were cultured together with autologous CD59 + or CD59 - lymphocytes, their cultured supernatant, extrinsic IFN γ and IL 2 in a liquid culture system. CD59 + cells were counted by flow cytometry after 10 day culture in vitro. The results showed that there were no differences in the proliferation ability of lymphocytes between each group: controls 1.42±0.46, unsorted PNH lymphocytes 1.40±0.35, CD59 - 1.30±0.40, and CD59 + PNH lymphocytes 1.40±0.42. Anti tumor effect of lymphocytes declined in PNH patients when compared with control [(50.00±28.67)% vs. (76.13±10.15)%]. The proportion of CD59 + cells diminished significantly after culture with autologous lymphocytes, their supernatant, extrinsic IFN γ or IL 2(P<0.01) in groups of unsorted, CD34 + and CD34 - bone marrow cells. No significant difference was found between groups of CD59 - and CD59 + lymphocytes, or CD34 - and CD34 + marrow cells. It is concluded that turbulences of immune regulations may be involved in pathogenesis of PNH.

关 键 词：阵发性睡眠性血红蛋白尿症 淋巴细胞 骨髓细胞 

分 类 号：R556.64[医药卫生—血液循环系统疾病]