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目的通过观察大剂量化疗(HDC)并自体外周血干细胞移植(APBSCT)治疗重症原发性干燥综合征(pSS)患者治疗前后 T 细胞亚群和 T 细胞受体(TCR)克隆谱型的变化,探讨移植的免疫重建过程。方法 2例长期糖皮质激素和免疫抑制剂治疗病情不能缓解的重症 pSS 患者,给予HDC 及 APBSCT 治疗。流式细胞仪检测 T 细胞亚群,观察治疗前后 T 细胞亚群的变化。RT-PCR 扩增患者外周血 TCRV 区24个β亚家族序列,阳性的亚家族行荧光标记的二次 PCR,用分析 TCRVβ亚家族 CDR3长度的方法检测 T 细胞克隆性,以及移植前后 T 细胞克隆谱的变化。结果 2例重症 pSS患者顺利完成 APBSCT 全过程,T 细胞亚群有不同程度的恢复,移植后 TCRVβ有由寡克隆向多克隆转化的趋势。结论本组2例重症 pSS 患者 HDC 并 APBSCT 治疗后病情得以控制,免疫功能在3个月时已有恢复的趋势,但其可行性与安全性有待大样本及随诊证实。
Objective To observe the changes of T cell subsets and T cell receptor (TCR) genotypes in patients with severe primary Sjogren’s syndrome (pSS) treated with high-dose chemotherapy (HDC) and autologous peripheral blood stem cell transplantation (APBSCT) Explore the immune reconstruction process. Methods Two long-term glucocorticoids and immunosuppressive agents were given HDC and APBSCT for patients with severe pSS who could not be alleviated. T-cell subsets were detected by flow cytometry and the changes of T-cell subsets were observed before and after treatment. RT-PCR amplification of peripheral blood TCRV 24 beta subfamily sequences, positive subfamily fluorescence labeling of the second PCR, TCRV beta subfamily CDR3 length analysis of T cell clonality and T cell cloning before and after transplantation Spectral changes. Results Two patients with severe pSS successfully completed the whole process of APBSCT. T cell subsets recovered to varying degrees. TCRVβ tended to be transformed from oligoclonal to polyclonal. Conclusions HDC and APBSCT were controlled in 2 patients with severe pSS in this group. The immune function has been restored at 3 months. However, its feasibility and safety need to be confirmed by large sample and follow - up.