摘要
目的 研究骨髓单个核细胞（BMMNC）-Coombs试验（＋）血细胞减少症（又称免疫相关性血细胞减少症,IRP）患者骨髓Th17细胞数量及功能,探讨Th17细胞在IRP发病中的作用.方法 选取初诊IRP患者43例、血常规恢复正常IRP患者34例及正常对照13名.采用流式细胞术（FCM）检测骨髓白细胞介素（IL）-23受体（IL-23R）＋CD4＋/CD4＋细胞比例;酶联免疫吸附（ELISA）法测定骨髓上清液IL-6、IL-23和IL-17水平;RT-PCR技术检测BMMNC视黄酸相关孤核受体γt（RORγt）及信号转导转录激活子3（STAT3）mRNA的相对表达水平.结果 IRP初诊组IL-23R＋CD4＋/CD4＋细胞比例[（3.6±3.1）%]明显高于恢复组[（2.0±1.0）%]和正常对照组[（1.9±1.4）%]（P值均＜0.05）;初诊组骨髓上清液IL-6、IL-23和IL-17水平[分别为（26.21±14.55）、（2.23±0.99）和（2.54±1.33）μg/L]明显高于恢复组[分别为（9.08±6.36）、（0.91±0.76）和（1.28±0.18）μg/L]和正常对照组[分别为（14.63±7.66）、（1.19±0.98）和（1.50±0.28）μg/L]（P值均＜0.05）;初诊组RORγtmRNA、STAT3mRNA表达水平（0.25±0.08、1.10±0.16）明显高于恢复组（0.12±0.08、0.97±0.12）和正常对照组（0.07±0.05、0.95±0.13）（P值均＜0.05）.IRP组IL-23R^＋CD4^＋/CD4^＋细胞比例与CD5^＋CD19^＋/CD19^＋细胞比例呈正相关（P＜0.05）;IRP组骨髓液IL-17水平与骨髓CD5^＋CD19^＋/CD19^＋细胞比例、自身抗体数量呈正相关（r分别为0.494、0.377,P值均＜0.05）;IRP组RORγt mRNA表达水平与骨髓CD5^＋CD19^＋/CD19^＋细胞比例及自身抗体数量呈正相关（r分别为0.741、0.541,P值均＜0.05）;IL-23R^＋CD4^＋/CD4^＋细胞比例与骨髓上清液IL-17水平及STAT3 mRNA表达水平呈正相关（r分别为0.438、0.448,P值均＜0.05）.结论 Th17细胞在IRP患者中数量增加,功能活跃;Th17细胞有望成为该类疾病干预治疗的新靶点.
Objective To study the quantity and function of bone marrow（BM）Th17 cells in the blood cytopenia patients with positive BMMNC-Coombs test（IRP）and to explore the role of Th17 cells in the pathogenesis of the disease.Methods Forty-three untreated IRP patients,34 recovered IRP patients and 13healthy donors were enrolled in this study.The ratio of IL-23R^＋ CD4^ ＋/CD4^＋ cells in BM were examined by flow cytometry（FCM）,the levels of IL-6,IL-23,IL-17 by ELISA,and the expressions of RORγt mRNA、STAT3 mRNA in BMMNC by semiquantitive RT-PCR.Results The ratio of IL-23R^＋ CD4^＋/CD4^＋ cells [（3.6±3.1）%],the levels of IL-6[（26.21 ±14.55）μg/L],IL-23[（2.23 ±0.99）μg/L],IL-17[（2.54 ± 1.33）μg/L]and the expressions of RORγt mRNA（0.25 ±0.08）and STAT3 mRNA（1.10 ±0.16）in BMMNC of untreated IRP patients were significantly higher than those of recovered IRP patients [（2.0±1.0）%,（9.08 ±6.36）μg/L,（0.91 ±0.76）μg/L,（1.28 ±0.18）μg/L,0.12 ±0.08,0.97 ±0.12 respectively]（P 〈0.05）;there was no significiant difference between those of recovered IRP patients and normal controls [（1.9±1.4）%, （14.63±7.66）μg/L, （1.19±0.98）μg/L, （1.50±0.28）μg/L, 0.07 ±0.05,0.95 ±0.13, respectively] （P 〉 0.05 ） . In IRP group, there were significantly positive correlations between the ratios of IL-23R ^＋ CD4^ ＋/CD4^ ＋ cells and CD5^ ＋ CD19^ ＋/CD19 ^＋ （P 〈 0.05 ）, there were significantly positive correlations between the levels of IL-17 and CD5^＋ CD19^＋/CD19^＋, the quantity of BMMNC-antibody （r =0.494 and 0. 377, respectively）（ P 〈 0.05 ）; and so did between the expressions of RORγt mRNA and the ratio of CD5 ^＋ CD19 ^＋/CD19 ^＋ , the quantity of BMMNC-antibody （ r = 0. 741 and 0. 541, respectively）（ P 〈0.05 ）, and between the ratio of IL-23R^＋ CD4^ ＋/CD4 ^＋ and the level of IL-17, the expression ofSTAT3 mRNA （r=0.438 and 0.448, respectively）（P〈0.05）. Conclusions There exists increased qunantity and hyperfunction of Th17 cells in the IRP patients which induce B cells hyperfunction and produetion of autoantibodies against the BM hematopoietic cells. Th17 cells might be a potential new therapeutic target of IRP.
出处
《中华血液学杂志》
CAS
CSCD
北大核心
2010年第10期684-687,共4页
Chinese Journal of Hematology
基金
国家自然科学基金（30670886,30470749,30971285,30971286）
天津市高等学校科研基金（200317）
天津市卫生局科技基金项目（03KZ26）
国家＂十一五＂科技支撑项目（2008BAI61B00）
高等学校博士学科点专项科研基金（200800620004）
天津市科技支撑计划重点项目（07ZCGYSF00600）
天津市应用基础及前沿技术研究计划（08JCYBJC07800）
