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目的研究骨髓增生异常综合征(MDS)患者骨髓 T 细胞极化状况及其与骨髓上清液中促凋亡因子 TNF-α、INF-γ水平及造血细胞凋亡的关系。方法采用流式细胞术检测34例 MDS 患者骨髓 T 细胞极化亚群状况,用 ELISA 法检测骨髓上清液 TNF-α、INF-γ水平,并用 TdT 介导的 dUTP 缺口末端标记法(TUNEL)检测骨髓造血细胞凋亡状况,分析三者间的相关性。结果①MDS 患者骨髓Th1细胞百分比[(10.1±1.6)%]和 Tc1细胞百分比[(24.0±3.6)%]以及 Tc1/Tc2值(50.0±11.1)明显高于正常对照[分别为(4.0±0.5)%,(5.8±0.6)%和13.4±2.7];MDS 患者骨髓上清液 INF-γ浓度[(58.6±21.7)μg/L]和 TNF-α浓度[(157±38)μg/L]较正常对照[分别为0和(0.3±0.2)μg/L]明显升高;MDS 患者骨髓造血细胞凋亡指数[(7.8±1.5)%]明显高于正常对照[(2.1±0.3)%,P<0.05]。骨髓 Th1细胞数与骨髓上清液 INF-γ和 TNF-α浓度呈正相关(r 值分别为0.38和0.39,P<0.05),并与造血细胞凋亡指数呈正相关(r=0.33,P<0.05)。骨髓上清液 INF-γ和 TNF-α浓度与造血细胞凋亡也呈正相关(r 值分别为0.74和0.73,P<0.01)。②难治性血细胞减少伴多系发育异常(RCMD)组(18例)Th1、Tc1细胞百分比,Tc1/Tc2值较正常对照明显升高(P 值均<0.01),T细胞向Ⅰ型反应极化;RCMD 伴环形铁粒幼细胞增多(RCMD-RS)(4例)、RAEB-1(5例)、RAEB-2组(7例)T 细胞未显示异常极化。③IPSS 低危组病例 Th1细胞百分比、Tc1细胞百分比及 Tc1/Tc2值均较正常升高,而高危组 T 细胞未见异常极化。④染色体核型正常的 RCMD 患者骨髓 Th1细胞百分比、Tc1细胞百分比及 Tc1/Tc2值均较正常对照明显升高。而核型异常组 T 细胞未见异常极化。结论MDS 患者骨髓中 Th1、Tc1细胞比例增高,Th1/Th2、Tc1/Tc2平衡向Ⅰ型反应极化,其中以 RCMD 组、IPSS 低危组和染色体核型正常组患者的 T 细胞Ⅰ型反应极化明显。骨髓造血细胞过度凋亡与 Th1细胞以及 INF-γ和 TNF-α水平呈正相关,提示 Th1细胞通过分泌 INF-γ和 TNF-α导致造血细胞过度凋亡。
Objective To investigate the bone marrow T cell polarization in patients with myelodysplastic syndrome (MDS) and its relationship with the levels of pro-apoptotic factors TNF-α and INF-γ in bone marrow supernatant and hematopoietic cell apoptosis. Methods The bone marrow T cell polarization subsets of 34 patients with MDS were detected by flow cytometry. The levels of TNF-α and INF-γ in bone marrow supernatants were detected by ELISA. TUNEL and TdT-mediated dUTP nick end labeling (TUNEL) Detection of bone marrow hematopoietic cells apoptosis, analysis of the correlation between the three. Results ① The percentages of Th1 cells and the percentage of Tc1 cells [(24.0 ± 3.6)%] and Tc1 / Tc2 (50.0 ± 11.1) in MDS patients were significantly higher than those in healthy controls [4.0 ± 0.5% %, (5.8 ± 0.6)%, and 13.4 ± 2.7] respectively. The concentrations of INF-γ in bone marrow supernatant of MDS patients [(58.6 ± 21.7) μg / L and The apoptotic index of bone marrow hematopoietic cells of MDS patients [(7.8 ± 1.5)%] was significantly higher than that of the normal controls [(2.1 ± 0.3)%, P <0.05]. The number of Th1 cells in bone marrow was positively correlated with the concentrations of INF-γ and TNF-α in bone marrow supernatant (r = 0.38 and 0.39, respectively, P <0.05), and positively correlated with hematopoietic apoptosis index (r = 0.33, ). The concentrations of INF-γ and TNF-α in bone marrow supernatant were also positively correlated with hematopoietic apoptosis (r = 0.74 and 0.73, respectively, P <0.01). ② The percentage of Th1 and Tc1 cells and Tc1 / Tc2 in RCMD group were significantly higher than those in normal control group (all P <0.01) RCMD with RCMD-RS (4 cases), RAEB-1 (5 cases) and RAEB-2 group (7 cases) T cells showed no abnormal polarization. ③ The percentages of Th1 cells, the percentage of Tc1 cells and the Tc1 / Tc2 values in low-risk PSS patients were higher than those in normal controls, while those in high-risk patients showed no abnormal polarization. ④ The percentages of Th1 cells, Tc1 cells and Tc1 / Tc2 in RCMD patients with normal karyotype were significantly higher than those in normal controls. However, there was no abnormal polarization of T cell in abnormal karyotype. Conclusion The proportion of Th1 and Tc1 cells in bone marrow of patients with MDS is increased, and the balance of Th1 / Th2 and Tc1 / Tc2 is polarized towards type Ⅰ reaction. The type Ⅰ response of T cell in RCMD group, IPSS low risk group and normal karyotype group Clearly. Over-apoptosis of bone marrow hematopoietic cells was positively correlated with Th1 cells and the levels of INF-γ and TNF-α, suggesting that Th1 cells induce excessive apoptosis of hematopoietic cells by secreting IFN-γ and TNF-α.