【摘要】 目的:建立多参数流式检测胸腺细胞选择相关高迁移率族蛋白(TOX)在PD-1、Tim-3和CD244耗竭性T细胞中的免疫表型法，以及其在1例罕见CD4~-CD56~+表型的血液恶性肿瘤母细胞性浆细胞样树突状细胞肿瘤(BPDCN)患者中的应用分析。方法:收集1例BPDCN患者骨髓(BM)和外周血(PB),2例非肿瘤手术患者和1例健康者BM以及11例健康者(HI)PB分别作为BM和PB对照组，利用多色流式单抗CD45、CD3、CD4、CD8、CD25、FoxP3、TOX、PD-1、Tim-3、CD244及相应同型对照进行流式细胞检测。结果:流式检测结果提示，BPDCN患者BM和PB中TOX~+CD3~+、TOX~+CD4~+和TOX~+CD8~+ T细胞比例均比对照组升高；BPDCN患者BM和PB中TOX~+PD-1~+CD8~+、TOX~+Tim-3~+CD8~+和TOX~+CD244~+CD8~+耗竭性T细胞中的分布比例也显著高于对照组。此外，在BPDCN患者BM和PB的CD4~+CD25~+FoxP3~+Treg细胞中TOX~+细胞分布比例高于对照组；同时TOX~+细胞在PD-1~+ CD4~+CD25~+FoxP3~+Treg、Tim-3~+ CD4~+CD25~+FoxP3~+Treg和CD244~+ CD4~+CD25~+FoxP3~+Treg耗竭性T细胞中的分布比例也明显高于对照组。结论:成功建立多参数流式细胞术检测PD-1、Tim-3和CD244耗竭性T细胞亚群中TOX免疫表型法，TOX可能是BPDCN患者T细胞免疫调节的重要转录因子。更多还原

【Abstract】 Objective:To establish a multi-parameter flow cytometry method for detecting the immunophenotypes of thymocyte selection-associated high mobility group box protein(TOX) expression and co-expression with programmed death 1(PD-1), T cell immunoglobulin domain and mucin domain-containing molecule 3(Tim-3) and CD244 in T cell subsets, and analyze the immune suppression characteristic in a patient with blastic plasmacytoid dendritic cell neoplasm(BPDCN). Methods: The peripheral blood(PB) and bone marrow(BM) of one patient with BPDCN were collected, while BM of non-tumor surgery(2 cases and 1 healthy donor), PB of 11 healthy individuals severed as control. The monoclonal antibodies CD45, CD3, CD4, CD8, CD25, FoxP3, TOX, PD-1, Tim-3, CD244 and corresponding isotype controls were employed in polychromatic flow cytometry detection. Results: The proportion of TOX~+CD3~+, TOX~+CD4~+ and TOX~+CD8~+ T cells in BM and PB of the patient with BPDCN was higher than that of healthy control. Moreover, the percentage of TOX~+PD-1~+CD8~+, TOX~+Tim-3~+CD8~+ and TOX~+CD244~+CD8~+ cells in BM or PB from the BPDCN patient was also higher than that from control. In addition, an increased percentage of TOX~+ cells among CD4~+CD25~+FoxP3~+ Treg cells in BM or PB of the BPDCN patients was observed in comparison with healthy individuals. And the number of TOX~+ cells among the PD-1~+CD4~+CD25~+FoxP3~+ Treg, Tim-3~+CD4~+CD25~+FoxP3~+ Treg and CD244~+CD4~+CD25~+FoxP3~+ Treg was also higher than that of healthy individuals. Conclusion:The detection of TOX expression and co-expression with PD-1, Tim-3 and CD244 in T cell subsets are successfully established using multi-parameter flow cytometry, and TOX may be an important transcription factor in T cell immune regulation of BPDCN patient.更多还原