阵发性睡眠性血红蛋白尿症异常克隆特征研究  

The Biological Characteristics of PNH Clones

作　　者：叶金慧 王璇 王世充 彭雪梅 葛美丽 邵英起 郑以州 程涛 胡林萍 YE Jinhui;WANG Xuan;WANG Shichong;PENG Xuemei;GE Meili;SHAO Yingqi;ZHENG Yizhou;CHENG Tao;HU Linping(Institute of Hematology&Blood Diseases Hospital,Haihe Laboratory of Cell Ecosystem,State Key Laboratory of Experimental Hematology,National Clinical Research Center for Blood Diseases,Chinese Academy of Medical Sciences&Peking Union Medical College,Tianjin 300020,China)

机构地区：[1]中国医学科学院血液病医院(中国医学科学院血液学研究所),细胞生态海河实验室,实验血液学国家重点实验室,国家血液系统疾病临床医学研究中心,中国医学科学院&北京协和医学院,天津300020

出　　处：《中国细胞生物学学报》2022年第7期1349-1358,共10页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:81970104、81890990);细胞生态海河实验室创新项目(批准号:HH22KYZX0040);国家重点研发计划(批准号:2021YFA1100900);中国医学科学院医学与健康科技创新工程项目(批准号:2021-I2M-1-040);中国医学科学院基金(批准号:2021-RW320-011)资助的课题。

摘　　要：阵发性睡眠性血红蛋白尿症(PNH)是一类源于造血干细胞PIGA基因突变的获得性克隆性疾病,临床表现主要为溶血、骨髓衰竭等。该文通过流式细胞术和体外集落形成等实验检测比较经典型PNH(cPNH)患者与健康对照组外周血细胞、骨髓中血细胞及造血干祖细胞(hemopoietic stem progenitor cells,HSPCs)的数量和功能,探究PNH异常克隆特征。结果显示,cPNH患者外周血细胞中T细胞、B细胞和NK细胞数明显降低,髓系细胞数无明显改变。骨髓各类血细胞中,cPNH患者单核细胞比例明显高于对照组,中性粒细胞、有核红细胞以及淋系细胞比例无明显改变。cPNH患者骨髓HSPCs均为CD59–的PNH异常克隆;患者HSPCs出现大量耗竭,向B-NK祖细胞分化明显受阻,该结果与外周血和骨髓中B、NK细胞变化一致,提示cPNH患者淋系细胞受损最为严重。CFU实验结果表明,cPNH患者HSPCs形成髓系克隆的能力与健康对照组无差异;而在红系克隆种类中,cPNH患者形成的偏小的红系克隆代偿性增多。综上,该研究解析了PNH异常克隆的生物学特征,为PNH的诊治提供了一定的理论依据。PNH(paroxysmal nocturnal hemoglobinuria)is an acquired clonal disease caused by the PIGA mutation in HSCs(hematopoietic stem cells),characterized mainly by hemolysis and bone marrow failure etc.In this study,flow cytometry and colony forming cell assays were performed to analyze the total number of blood cells and HSPCs in bone marrow and peripheral blood from cPNH patients compared with health donors.Moreover,the characteristics of PNH clones were investigated.The results revealed that the number of T cells,B cells and NK cells in peripheral blood from cPNH decreased significantly,while no significant difference was observed in the number of myeloid cells compared with health donors.Among many kinds of blood cells in bone marrow,the proportion of monocytes in cPNH was significantly higher than that in the control group,while there was no significant difference in the proportion of neutrophils,nucleated erythrocytes and lymphoid cells between cPNH and control group.All the HSPCs from cPNH bone marrow were abnormal PNH clones detected as CD59–.Moreover,there was an extensive exhaustion of HSPCs in cPNH,and their lymphoid differentiation potential was obviously inhibited,which was consistent with the change of B cells and NK cells in peripheral blood and bone marrow.These data indicated that lymphoid cells were the most severely impaired cells in cPNH.CFU assays manifested that there was no difference in myeloid colony forming ability between control and cPNH.As for the erythroid colony,cPNH HSPCs formed more small erythroid colonies for compensation.In conclusion,this study showed the biological characteristics of PNH clones,which provided theoretical basis for the diagnosis and treatment of PNH.

关 键 词：阵发性睡眠性血红蛋白尿症 PNH克隆 特征 造血干祖细胞 

分 类 号：R556.64[医药卫生—血液循环系统疾病]