摘要
目的探讨原发免疫性血小板减少症（ITP）患者外周血CD5＋B细胞数量及其分泌IL．10的能力及意义。方法采用流式细胞术检测57例治疗前、40例治疗后IrrP患者及25名正常对照外周血CD5＋B细胞比例及其在体外培养后胞内IL-10的平均荧光强度（MFI），ELISA方法检测培养上清液IL．10浓度；分析CD5＋B细胞特征与疾病状态的关系。结果治疗前ITP患者外周血CD5＋B细胞占总淋巴细胞比例和绝对值计数[（3．75±2．37）％、（6．29±5．77）×10＋／L]均高于正常对照[（2．10±1．08）％、（3．06±1．90）×10＋／L]（P值均〈0．05）；ITP患者和正常对照外周血CD5＋B细胞的胞内IL-10的MFI水平均高于其他淋巴细胞亚群；治疗前ITP患者CD5‘B细胞中产生IL-10的细胞比例、胞内IL-10MFI[（29．51±20．73）％、27．95±13．99]均高于正常对照[（15．904-9．58）％、14．31±11．29]（P值均〈0．05），其细胞培养上清液中IL-10浓度[（173．05±102．50）ng／L]低于正常对照组[（230．61±76．96）ng／L]；治疗后ITP患者CD5＋B细胞比例下降至正常水平，其胞内IL．10MFI与治疗前无显著差异，仍高于正常对照组，但细胞培养上清液中IL-10水平恢复正常。结论治疗前ITP患者外周血CD5＋B细胞比例升高、细胞内IL-10积聚，但分泌IL-10的能力下降；疾病缓解后，CD5＋B细胞的数量及分泌IL-10的能力得到恢复。
Objective To investigate the number of peripheral blood CD5 B cells and their ability ＋ of secreting IL-10 in patients with immune thrombocytopenia（ITP）. Methods Peripheral blood lymphocytes were isolated from 57 pre-treated, 40 post-treated ITP patients and 25 controls using Ficoll-Hypaque density centrifugation and then stained with PE-CD5/FITC-CD19 for flow cytometric analysis. After 24-hour culture, lymphocytes were stained with APC-IL-10 for intracellular cytokine detection. ELISA assay was employed to determine IL-IO concentration in supernatants. Results The percentage and absolute number of CD5＋ B cells in peripheral blood from pre-treated ITP patients were significantly higher than that from normal controls （3.75±2.37）% vs （2.10±1.08）%, P〈0.01;（6.29±5.77） xlOT/Lvs（3.06±1.90） x 10V/L,P〈 0.01. CD5＋ B cells expressed more intracellular IL-10 than other lymphocyte subsets both in ITP patients and normal controls. The percentages of IL-10 cells within CD5 ＋ B cells in pre-treated ITP patients and normal controls were （ 29.51 ±20.73 ） % and （ 15.90 ± 9.58 ） %, respectively （ P 〈 0.01 ）. Intracellular mean fluo- rescence intensity （MFI） of IL-IO in CD5 ＋ B cells was 27.95± 13.99 in pre-treated patients, which was sig- nificantly higher than that in controls （ P 〈 0.01 ）. In contrast, IL-10 concentration in supernatants was （173.05 ±102.50）ng./L in pre-treated ITP group, which was lower than that（230.61± 76.96）ng/L in con- trois. In patients who achieved remission, the number of CD5 ＋ B cells decreased to level comparable to nor- real controls. While intracellular IL-10 MFI of CD5 ＋ B cells in post-treated ITP patients remained as high as in pre-treated ones, the IL-10 concentration in supernatants increased to level similar to controls. Conclusion The significantly increased number of CD5 ＋ B cells and accumulated IL-10 in CD5 ＋ B cells suggestedimpaired IL-IO secretion in ITP patients. The number and the ability of secreting
出处
《中华血液学杂志》
CAS
CSCD
北大核心
2012年第12期1028-1032,共5页
Chinese Journal of Hematology
基金
国家自然科学基金（30972737、81170473）
上海市卫生局科研计划课题（20114313）
