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