[1]王国征,吴远彬,郭坤元,等.CD34CD38KG1a白血病干细胞对同种异体自然杀伤细胞的抵抗作用及其机制[J].新乡医学院学报,2019,36(9):828-832.[doi:10.7683/xxyxyxb.2019.09.006]
　WANG Guo-zheng,WU Yuan-bin,GUO Kun-yuan,et al.Resistance of CD34CD38KG1a leukemic stem cells to allogeneic nature killer cells and its mechanism[J].Journal of Xinxiang Medical University,2019,36(9):828-832.[doi:10.7683/xxyxyxb.2019.09.006]
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CD34+CD38-KG1a白血病干细胞对同种异体自然杀伤细胞的抵抗作用及其机制
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
36
期数:
2019年9
页码:
828-832
栏目:
基础研究
出版日期:
2019-09-05

文章信息/Info

Title:
Resistance of CD34+CD38-KG1a leukemic stem cells to allogeneic nature killer cells and its mechanism
作者:
王国征; 吴远彬; 郭坤元; 代喜平
（广州中医药大学第二附属医院血液科，广东省中医院血液科，广东　广州　510120）
Author(s):
WANG Guo-zheng; WU Yuan-bin; GUO Kun-yuan; DAI Xi-ping
（Department of Hematology，the Second Affiliated Hospital of Guangzhou University of Chinese Medicine；Guangdong Provincial Hospital of Chinese Medicine，Guangzhou 510120，Guangdong Province，China）
关键词:
CD34+CD38-KG1a细胞; 自然杀伤细胞; 细胞毒实验; NKG2D配体
Keywords:
CD34+CD38-KG1a cell; nature killer cell; cytotoxicity; NK2D ligand
分类号:
R733.71
DOI:
10.7683/xxyxyxb.2019.09.006
文献标志码:
A
摘要:
目的　探讨CD34+CD38-KG1a白血病干细胞抵抗同种异体自然杀伤（NK）细胞杀伤作用的机制。方法　采用免疫磁珠法（MACS）分选CD34+CD38-KG1a细胞和4例健康个体外周血中NK细胞，并采用流式细胞术检测纯度；乳酸脱氢酶（LDH）释放法检测NK细胞在不同效靶比（5∶1，10∶1，20∶1）下对K562和CD34+CD38-KG1a细胞的杀伤作用；聚合酶链式反应-序列特异性引物法分析NK细胞KIR和CD34+CD38- KG1a细胞HLA-Ⅰ基因分型；流式细胞术检测K562和CD34+CD38-KG1a细胞表面主要组织相容性复合体（MHC）-I类链相关分子A/B、人巨细胞病毒 UL16结合蛋白（ULBP）1～3和人类白细胞抗原（HLA）-Ⅰ类分子的表达。结果　分选的CD34+CD38--KG1a细胞和NK细胞纯度分别为（94.25±2.16）%、（91.70±2.05）%。NK细胞在各效靶比下对CD34+CD38-KG1a细胞的杀伤率均低于K562细胞（P<0.05）。4例健康个体NK细胞KIR基因型为KIR2DL1、KIR2DL3、KIR3DL1和KIR3DL2，CD34+CD38-KG1a细胞HLA-Ⅰ基因型为A30、30，B51、78，Cw4、16。K562细胞高表达NKG2D配体，CD34+CD38-KG1a细胞几乎不表达NKG2D配体，CD34+CD38-KG1a细胞中NKG2D配体表达率显著低于K562细胞（P<0.05）；K562细胞中几乎不表达HLA-Ⅰ，CD34+CD38-KG1a细胞高表达HLA-Ⅰ，CD34+CD38-KG1a细胞中HLA-Ⅰ的表达率显著高于K562细胞（P<0.05）。结论　CD34+CD38-KG1a细胞明显抵抗同种异体NK细胞的杀伤作用，其机制可能与高表达HLA-Ⅰ分子和低表达NK2D配体有关。
Abstract:
Objective　To explore the mechanism of CD34+CD38-KG1a leukemia stem cells against killing effect of nature killer （NK）cells.Methods　The CD34+CD38-KG1a cells and NK cells of 4 healthy individuals were isolated by magnetic cell sorting （MACS） and the purity were detected by flow cytometry.The killing effects of NK cells on K562 and CD34+CD38-KG1a cells at different effector-to-target cell ratios（5∶1，10∶1，20∶1） were detected by lactate dehydrogenase （LDH） release assay.KIR genotyping of NK cells and HLA-I genotyping of CD34+CD38-KG1a cells were analyzed by polymerase chain reaction-sequence specific primers.The expression of major histocompatibility complex（MHC） class I chain-related molecules A and B（MICA/B），human cytomegalovirus glycoprotein UL16 binding proteins（ULBP）1-3 and human leukocyte antigen（HLA）-I molecules on the surface of K562 and CD34+CD38-KG1a cells were detected by flow cytometry.Results　The purity of CD34+CD38-KG1a cells and NK cells were （94.25+2.16）% and （91.70+2.05）% respectively.The killing rate of NK cells to CD34+CD38- KG1a cells was lower than that to the K562 cells under all effector-to-target cell ratios （P<0.05）.The KIR genotypes of NK cells in 4 healthy individuals were KIR2DL1，KIR2DL3，KIR3DL1 and KIR3DL2；the HLA-I genotypes of CD34+CD38-KG1a cells were A30，30；B51，78；Cw4，16.NKG2D ligand was highly expressed in K562 cells，and NKG2D ligand was almost not expressed in CD34+CD38-KG1a cells；the expression rate of NKG2D ligand in CD34+CD38-KG1a cells was significantly lower than that in the K562 cells （P<0.05）.The HLA-I was almost not expressed in K562 cells，but it was highly expressed in CD34+CD38-KG1a cells；the expression rate of HLA-I in CD34+CD38-KG1a cells was significantly higher than that in the K562 cells（P<0.05）.Conclusion　CD34+CD38-KG1a cells can significantly resistant the killing of allogeneic NK cells，which may be related to the high expression of HLA-I and the low expression of NK2D ligands.

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更新日期/Last Update: 2019-09-05