摘要
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 explore polarization of T lymphocyte and its relationship with apoptosis of marrow cells in patients with myelodysplastie syndromes （MDS）. Methods Measurements of Thl, Th2, Tc1, Tc2 subsets in bone marrows from 34 patients with MDS and 13 normal controls were performed by flow eytometry. INF-γ and TNF-α in marrow serum were determined by ELISA （ Enzyme-linked immunosorbent assay）. Apoptosis index of marrow cells was detected by TUNEL （TdT-mediated dUTP nick end labeling）. Correlations between Th1, Th2, Tc1, Tc2 subsets and INF-γ, TNF-α levels as well as apoptosis index were analyzed, and relationship between TNF-α, INF-γ levels and apoptosis index was also investigated. Results ①The percentage of Thl cells [（10. 1 ±1.6）%], Tel cells [（24.0 ±3.6）%] and Tc1/Tc2 ratio （50.0 ± 11.1 ） was significantly increased in patients with MDS than in normal controls [ （4.0 ± 0. 5 ） % , （5.8 ±0.6）% and 13.4 ±2.7, respectively]. Levels of INF-γ [ （58.6 ±21.7）μg/L] and TNF-α [ （15.7±3.8） μg/L] in marrow serum of MDS patients was markedly elevated compared to normal controls [0 and （0.3 ± 0.2） μg/L, respectively ]. An increased apoptosis index of nucleated cells was observed in MDS patients [（7.8±1.5）%] as compared to controls [（2. 1 ±0.3）%, P〈0.05]. The Thl cell percentage showed a positive correlation with the levels of INF-γ, and TNF-α （ r = 0. 38, P 〈 0.05 and r = 0. 39, P 〈 0.05, respectively）, and with apoptotic index of nucleated marrow cells in MDS patients （ r = 0. 33, P 〈 0.05 ）. Furthermore, a positive correlation was observed between INF-γ, TNF-α levels and apoptotic index of marrow cells （r =0. 74, P 〈0.01 and r =0. 73, P 〈0.01, respectively）. ①Th1, Tc1 cells and Tc1/Tc2 ratio in MDS-RCMD patients was markedly elevated （P 〈 0.01 ） but did not in RCMD-RS, RAEB-1 and RAEB-2 patients as compared to normal controls. ③An elevation in the percentages of Thl, Tcl and Tc1/ Tc2 ratio was detected in patients with IPSS lower-risk but did not in higher-risk group as compared to controls. ④Increased Thl and Tel percentages and Th1/Th2 and Tc1/Tc2 ratios were observed in RCMD patients with normal karyotype, but did not in those with abnormal karyotype. Conclusions Th1/Th2 and Tc1/Tc2 in bone marrow of MDS patients were unbalanced, polarizing to type I reaction especially in patients with RCMD subtype, IPSS lower-risk and normal karyotype. The increased Thl cells in bone marrow may account for the increased apoptosis of nucleated marrow cells in MDS, through proapoptotic cytokines such as INF-γ, and TNF-α. [
出处
《中华血液学杂志》
CAS
CSCD
北大核心
2007年第8期549-554,共6页
Chinese Journal of Hematology
