Abstract
目的探索再生障碍性贫血(AA)小鼠模型中供者来源T细胞不同时间点动力学变化。方法构建AA小鼠模型,分别于不同时间点采用流式细胞术测定模型小鼠脾脏与骨髓内供者T细胞比例、活化分子表达、细胞周期及功能亚群,评估不同时期T细胞功能状态。结果①半致死剂量照射联合主要组织相容性抗原(MHC)半相合的淋巴结细胞输注成功构建T细胞免疫介导的AA小鼠模型。②AA小鼠脾脏中供者T细胞从移植第3天后开始明显浸润,并逐渐出现CD4+/CD8+比例倒置,第5天开始进入骨髓,以CD8+细胞浸润为主。③CD69在供者CD4+细胞中表达高峰晚于CD8+细胞,CD25在CD4+细胞与CD8+细胞中表达水平的变化趋势相同,但在CD8+细胞中的表达高于CD4+细胞。④脾脏内供者CD4+细胞S/G2/M期比例在移植后第3天即达高峰,约12%,而CD8+细胞中S/G2/M期比例在移植后第5天达高峰,约20%,且两者在进入骨髓后S/G2/M期比例均再次升高,但移植第3天后其比例在脾脏与骨髓CD8+细胞中持续高于CD4+细胞。⑤脾脏内免疫活化的T细胞经历短暂的中枢记忆T细胞(TCM)阶段后迅速分化为效应记忆T细胞(TEM),进入骨髓后部分TEM分化为效应细胞进一步发挥效应功能。结论AA小鼠模型中供者T细胞进入异体后迅速活化,5天内达增殖高峰,并完成向TEM细胞的分化,5天后开始进入骨髓进一步增殖损伤造血。
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