CAJ | 학술논문

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ITP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)×107/L]均高于正常对照[(2.10±1.08)%、(3.06±1.90)×107/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-10 MFI[(29.51±20.73)%、27.95±13.99]均高于正常对照[(15.90±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-10 MFI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.
목적 탐토원발면역성혈소판감소증(ITP)환자외주혈CD5+ B세포수량급기분비IL-10적능력급의의.방법 채용류식세포술검측57례치료전、40례치료후ITP환자급25명정상대조외주혈CD5+ B세포비례급기재체외배양후포내IL-10적평균형광강도(MFI),ELISA방법검측배양상청액IL-10농도;분석CD5+ B세포특정여질병상태적관계.결과 치료전ITP환자외주혈CD5+ B세포점총림파세포비례화절대치계수[(3.75±2.37)%、(6.29±5.77)×107/L]균고우정상대조[(2.10±1.08)%、(3.06±1.90)×107/L](P치균＜0.05);ITP환자화정상대조외주혈CD5+ B세포적포내IL-10적MFI수평균고우기타림파세포아군;치료전ITP환자CD5+ B세포중산생IL-10적세포비례、포내IL-10 MFI[(29.51±20.73)%、27.95±13.99]균고우정상대조[(15.90±9.58)%、14.31±11.29](P치균＜0.05),기세포배양상청액중IL-10농도[(173.05±102.50)ng/L]저우정상대조조[(230.61±76.96)ng/L];치료후ITP환자CD5+ B세포비례하강지정상수평,기포내IL-10 MFI여치료전무현저차이,잉고우정상대조조,단세포배양상청액중IL-10수평회복정상.결론 치료전ITP환자외주혈CD5+ B세포비례승고、세포내IL-10적취,단분비IL-10적능력하강;질병완해후,CD5+ B세포적수량급분비IL-10적능력득도회복.
Objective To investigate the number of peripheral blood CD5+ B cells and their ability of secreting IL-10 in patients with immune thrombocytopenia(ITP). MethodsPeripheral 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-10 concentration in supernatants. ResultsThe 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）×107/L vs（3.06±1.90）×107/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 fluorescence intensity (MFI) of IL-10 in CD5+ B cells was 27.95±13.99 in pre-treated patients, which was significantly 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 controls. In patients who achieved remission, the number of CD5+ B cells decreased to level comparable to normal 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 suggested impaired IL-10 secretion in ITP patients. The number and the ability of secreting IL-10 of CD5+ B cells could be restored after effective treatments in patients with ITP.