Zhang Y J, Qu W, Liu H, Wang Y H, Liu C Y, Li L J, Wang H Q, Fu R, Xing L M, Shao Z H
Department of Hematology, Tianjin Medical University General Hospital, Tianjin 300052, China.
Zhonghua Yi Xue Za Zhi. 2017 Apr 25;97(16):1231-1235. doi: 10.3760/cma.j.issn.0376-2491.2017.16.010.
To investigate natural killer (NK) cell quantities and function in patients with immune thrombocytopenia (ITP) . A total of 66 ITP patients (34 newly diagnosed and 32 in complete remission) were collected from September 2015 to May 2016 in Tianjin Medical University General Hospital, and 30 healthy volunteers were recruited as controls. The percentages of NK cells and their subsets in peripheral blood, the expression of activating receptor (NKp44), inhibitory receptor (NKG2A) and CD16, perforin and granzyme β were detected by flow cytometry. The correlation between the above parameters and patients' immune status and platelet level were evaluated. (1)The percentage of CD3(-)CD56(+) NK cells in newly diagnosed patients (10.99%±4.89%)and patients in complete remission (9.73%±6.75%) were significantly lower than that in healthy controls (14.67%±7.24%)(=0.023, 0.003). The percentage of NK cells Bright subset was significantly lower in the newly diagnosed patients(0.48%±0.23%)and those in complete remission (0.41%±0.33%) than in healthy controls(0.64%±0.32%)(=0.037, 0.002); the percentage of Dim subset was also significantly lower in the newly diagnosed (10.16%±5.02%) and patients in complete remission (8.07%±5.74%) than in healthy controls(14.16%±7.19%) (=0.009, 0.007). (2)The proportion of Bright subset in total NK cells in new diagnosed ITP patients (6.48%±4.33%) was significantly higher than that in healthy controls (4.21%±2.70%)(=0.020); the proportion of Dim NK cells subset in new diagnosed ITP patients (93.51%±4.33%) was significantly lower than that in healthy controls(95.79%±2.70%) (=0.020). (3)The expression of activating receptor NKp44 in new diagnosed ITP patients was significantly lower than that in complete remission group and healthy controls[0.28%(0.95%)vs 0.61%(2.05%), 0.92%(0.90%); =0.047, 0.048]; the expression of inhibitory receptor NKG2A in new diagnosed ITP patients was significantly higher than that in healthy controls(42.34%±23.86% vs 29.25%±12.83%, =0.009). The proportion of CD16 was significantly lower in the newly diagnosed patients than in healthy controls(93.51%±4.33%95.79%±2.70%, =0.020). (4)The expression of perforin in the newly diagnosed ITP patients was significantly lower than that in healthy controls [87.52%(25.29%)vs 91.55%(8.29%), =0.025]; the expression of granzyme β in ITP patients and controls showed no statistically significant difference. (5)The level of NK cells in ITP patients was negatively correlated with CD3(+) CD8(+) T cells (=-0.387, =0.012) and CD5(+) CD19(+) B cells in peripheral blood (=-0.273, =0.028), positively correlated with the ratio of CD3(+) CD4(+) /CD3(+) CD8(+) (=0.358, =0.028) and peripheral platelet count (=0.314, =0.011). Deceased quantities and impaired total NK function, insufficient suppression of autoreactive T and B cells might play a role in the pathogenesis of ITP.
探讨免疫性血小板减少症(ITP)患者自然杀伤(NK)细胞数量及功能。2015年9月至2016年5月在天津医科大学总医院收集了66例ITP患者(34例新诊断患者和32例完全缓解患者),招募30名健康志愿者作为对照。采用流式细胞术检测外周血NK细胞及其亚群的百分比、活化受体(NKp44)、抑制性受体(NKG2A)和CD16、穿孔素及颗粒酶β的表达。评估上述参数与患者免疫状态及血小板水平的相关性。(1)新诊断患者(10.99%±4.89%)和完全缓解患者(9.73%±6.75%)中CD3(-)CD56(+)NK细胞的百分比显著低于健康对照(14.67%±7.24%)(P=0.023,0.003)。新诊断患者(0.48%±0.23%)和完全缓解患者(0.41%±0.33%)中NK细胞Bright亚群的百分比显著低于健康对照(0.64%±0.32%)(P=0.037,0.002);新诊断患者(10.16%±5.02%)和完全缓解患者(8.07%±5.74%)中Dim亚群的百分比也显著低于健康对照(14.16%±7.19%)(P=0.009,0.007)。(2)新诊断ITP患者中NK细胞Bright亚群在总NK细胞中的比例(6.48%±4.33%)显著高于健康对照(4.21%±2.70%)(P=0.020);新诊断ITP患者中Dim NK细胞亚群的比例(93.51%±4.33%)显著低于健康对照(95.79%±2.70%)(P=0.020)。(3)新诊断ITP患者中活化受体NKp44的表达显著低于完全缓解组和健康对照[0.28%(0.95%)对0.61%(2.05%),0.92%(0.90%);P=0.047,0.048];新诊断ITP患者中抑制性受体NKG2A的表达显著高于健康对照(42.34%±23.86%对29.25%±12.83%,P=0.009)。新诊断患者中CD16的比例显著低于健康对照(93.51%±4.33%对95.79%±2.70%,P=0.020)。(4)新诊断ITP患者中穿孔素的表达显著低于健康对照[87.52%(25.29%)对91.55%(8.29%),P=0.025];ITP患者和对照中颗粒酶β的表达无统计学差异。(5)ITP患者中NK细胞水平与外周血CD3(+)CD8(+)T细胞呈负相关(r=-0.387,P=0.012),与CD5(+)CD19(+)B细胞呈负相关(r=-0.273,P=0.028),与CD3(+)CD4(+)/CD3(+)CD8(+)比值呈正相关(r=0.358,P=0.028),与外周血小板计数呈正相关(r=0.314,P=0.011)。NK细胞数量减少及总功能受损、对自身反应性T和B细胞抑制不足可能在ITP发病机制中起作用。