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, General Hospital, Tianjin Medical University, Tianjin 300052, China.
Zhonghua Xue Ye Xue Za Zhi. 2017 May 14;38(5):399-403. doi: 10.3760/cma.j.issn.0253-2727.2017.05.009.
To investigate the levels of NK cells and their relevant cytokines (IL-10, TGF-β and IFN-γ) in patients with primary immune thrombocytopenia (ITP) . All samples were obtained from 42 patients (22 newly diagnosed and 20 in remission) and 20 healthy volunteers. The levels of IL-10 and IFN-γ in blood serum were detected by enzyme-linked immunosorbent assay (ELISA) . The percentage of CD3(-) CD56(+) NK cell, CD3(-) CD56(bright) CD16(-) NK cell, CD3(-) CD56(dim) CD16(+) NK cell in peripheral blood lymphocyte were detected by flow cytometry. The NK cells were isolated by immunomagnetic microbeads. The mRNA expression levels of IL-10, TGF-β, and IFN-γ in NK cells were detected by real-time fluorescent quantitative PCR. Correlation between the above measured results was analyzed. ① The blood serum level of IFN-γ in newly diagnosed ITP patients [ (653.0±221.6) ng/L] was higher than that in remission ITP patients [ (484.4±219.5) ng/L] and healthy control [ (390.9±253.5) ng/L] (=0.022, =0.001) . The blood serum level of IL-10 in newly diagnosed ITP patients was lower than that in healthy control [ (52.09±26.66) ng/L (79.44±38.43) ng/L, =0.007]. ②The percentage of NK cell in newly diagnosed and remission ITP patients [ (9.53±3.93) %, (9.03±3.78) %] were significantly lower than that in healthy control [ (13.72±7.42) %] (=0.013, =0.007) . The ratio of CD3(-) CD56(bright) CD16(-) NK cell/total NK cells in newly diagnosed ITP patients was higher than that in healthy control [ (6.85±4.43) % (4.05±2.81) %, =0.032]. The ratio of CD3(-)CD56(dim) CD16(-) NK cell/total NK cells in newly diagnosed ITP patients was lower than that in healthy control [ (93.14±4.43) % (95.94±2.81) %, =0.032]. ③ There was no significant difference in the mRNA expression level of IFN-γ in NK cells of ITP patients and healthy control (all >0.05) . The mRNA expression levels of IL-10 and TGF-β in NK cells in newly diagnosed ITP patients were significantly higher than that in healthy control (1.82±1.32 1.02±1.03, =0.023; 2.80±2.31 1.46±1.37, =0.028) . The ratio of CD3(-)CD56(bright) CD16(-) NK cell/total NK cells was positively correlated with the mRNA expression levels of IL-10, TGF-β in NK cells (=0.424, =0.001; =0.432, <0.001) . NK cells may compensate for the deficiency of the number by enhancing the secretion of negative regulation cytokines, acting as "protective" roles in the disease.
探讨原发性免疫性血小板减少症（ITP）患者自然杀伤（NK）细胞及其相关细胞因子（白细胞介素-10、转化生长因子-β和干扰素-γ）的水平。所有样本取自42例患者（22例新诊断患者和20例缓解期患者）及20名健康志愿者。采用酶联免疫吸附测定（ELISA）法检测血清中白细胞介素-10和干扰素-γ的水平。采用流式细胞术检测外周血淋巴细胞中CD3(-)CD56(+)NK细胞、CD3(-)CD56(bright)CD16(-)NK细胞、CD3(-)CD56(dim)CD16(+)NK细胞的百分比。通过免疫磁珠分离NK细胞。采用实时荧光定量聚合酶链反应检测NK细胞中白细胞介素-10、转化生长因子-β和干扰素-γ的信使核糖核酸（mRNA）表达水平。分析上述检测结果之间的相关性。①新诊断ITP患者血清干扰素-γ水平[（653.0±221.6）ng/L]高于ITP缓解期患者[（484.4±219.5）ng/L]和健康对照[（390.9±253.5）ng/L]（=0.022，=0.001）。新诊断ITP患者血清白细胞介素-10水平低于健康对照[（52.09±26.66）ng/L （79.44±38.43）ng/L，=0.007]。②新诊断和缓解期ITP患者NK细胞百分比[（9.53±3.93）%，（9.03±3.78）%]显著低于健康对照[（13.72±7.42）%]（=0.013，=0.007）。新诊断ITP患者CD3(-)CD56(bright)CD16(-)NK细胞与总NK细胞的比例高于健康对照[（6.85±4.43）% （4.05±2.81）%，=0.032]。新诊断ITP患者CD3(-)CD56(dim)CD16(-)NK细胞与总NK细胞的比例低于健康对照[（93.14±4.43）% （95.94±2.81）%，=0.032]。③ITP患者与健康对照NK细胞中干扰素-γ的mRNA表达水平无显著差异（均>0.05）。新诊断ITP患者NK细胞中白细胞介素-10和转化生长因子-β的mRNA表达水平显著高于健康对照（1.82±1.32 1.02±1.03，=0.023；2.80±2.31 1.46±1.37，=0.028）。CD3(-)CD56(bright)CD16(-)NK细胞与总NK细胞的比例与NK细胞中白细胞介素-10、转化生长因子-β的mRNA表达水平呈正相关（=0.424，=0.001；=0.432，<0.001）。NK细胞可能通过增强负调控细胞因子的分泌来弥补数量的不足，在该疾病中发挥“保护”作用。
