急性髓系白血病患者NK细胞表达及功能变化研究

The Expression and Function of NK Cells in Patients with Acute Myeloid Leukemia

目的:探讨急性髓系白血病(AML)患者自然杀伤(NK)细胞的抗原表达特征和功能标记物的表达特点。方法:采用多参数流式细胞术检测56例初诊AML患者临床样本以及24例健康对照样本中的NK细胞表面标记物以及其功能指标,包括激活性受体NKG2D、NKP46、DNAM-1和杀伤标记颗粒酶B、穿孔素。结果:56例AML患者参照WHO造血和淋巴组织肿瘤分类标准大致分为AML未分化型(M_(1))、AML部分分化型(M_(2))和急性粒/单核细胞白血病(M_(4)/M_(5))3类,但3类患者的NK细胞并无差异,因而不再细分。NK细胞分为CD3^(-)CD56^(hi)CD16^(-)(简称CD56^(hi)NK)和CD3^(-)CD56^(dim)CD16^(+)(简称CD56^(dim)NK)两个细胞群来分析。AML患者CD56^(hi)NK细胞群与CD56^(dim)NK细胞群相比,除NKP46外,CD56^(hi)NK细胞的NKG2D等受体阳性百分比均明显降低(P<0.001)。与健康对照相比,AML患者CD56^(hi)NK细胞比例明显增高,而NK细胞数量、比例和CD56^(dim)NK细胞比例明显降低(P<0.05);CD56^(hi)NK细胞胞内穿孔素分子比例明显增高(P<0.05);CD56^(hi)NK细胞表面DNAM-1分子,CD56^(dim)NK细胞表面NKG2D、DNAM-1分子以及胞内穿孔素分子表达明显减低(P<0.05)。AML患者其余功能指标表达与健康对照对应指标相比差异均无统计学意义(P>0.05)。另外,AML患者CD56^(hi)NK细胞比例与CD34^(+)表达呈正相关关系(r=0.303)。结论:与CD56^(dim)NK相比,AML患者的CD56^(hi)NK比例及功能标记物表达量都较低;与健康对照相比,AML患者的NK细胞数量及表达比例降低且功能标记物表达异常,提示其功能受损。

Objective:To explore the expression characteristics of antigens and functional markers of natural killer(NK)cells in patients with acute myeloid leukemia(AML).Methods:Multi-parameter flow cytometry was used to detect NK cell surface markers and their functional indicators in 56 newly diagnosed AML patients and 24 healthy controls,including activating receptors NKG2 D,NKP46,DNAM-1,and killing indicators granzyme B,perforin.Results:Referring to the WHO hematopoiesis and lymph tissue tumor classification criteria,56 cases were roughly divided into three types:AML M_(1),M_(2),and M_(4)/M_(5).However,there was no differences about NK cells among the three types,so it was no longer subdivided.NK cells were divided into two groups:CD3^(-)CD56^(hi)CD16^(-)(CD56^(hi)NK)and CD3^(-)CD56^(dim)CD16^(+)(CD56^(dim)NK).Compared with CD56^(dim)NK cell population,except for NKP46,the positive expression levels of NKG2 D and other receptors of CD56^(hi)NK cells in AML patients decreased(P<0.001).Compared with healthy controls,the proportion of CD56^(hi)NK cells in AML patients increased,while the number and proportion of NK cells and proportion of CD56^(dim)NK cells significantly decreased(P<0.05).The proportion of perforin in CD56^(hi)NK cells significantly increased(P<0.05).The expression of DNAM-1 in CD56^(hi)NK cells,NKG2 D,DNAM-1,and perforin in CD56^(dim)NK cells decreased significantly(P<0.05).There was no statistically significant difference in expression of other functional indexes in AML patients compared with corresponding indexes of healthy controls.In addition,the proportion of CD56^(hi)NK cells was positively correlated with the expression of CD34^(+) in AML(r=0.303).Conclusion:Compared with CD56^(dim)NK,the ratio of CD56^(hi)NK and the expression of functional markers in AML patients are lower.Compared with healthy controls,the number and expression ratio of NK cells in AML patients decrease and the expression of functional markers is abnormal,indicating that its function is impaired.