A mouse model of irradiation and spleen-thymus lymphocyte infusion induced aplastic anemia
A mouse model of irradiation and spleen-thymus lymphocyte infusion induced aplastic anemia
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DOI：
10.1080/16078454.2022.2113356
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发表时间：
2022-08
期刊：
影响因子：
1.9
通讯作者：
Runfeng Ni;Liwei Fan;Le Zhang;Yanqi Song;HaiJin Wang;Aidi Wang;Baoshan Liu
中科院分区：
文献类型：
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作者：
Runfeng Ni;Liwei Fan;Le Zhang;Yanqi Song;HaiJin Wang;Aidi Wang;Baoshan Liu
摘要目的建立免疫诱导的再生障碍性贫血（AA）小鼠模型，用于AA的研究。然而，没有统一的建立模型的条件，也没有评估免疫稳态。本研究的目的是确定AA模型的建立条件，并对AA模型进行免疫学和病理学评价。方法采用亚致死量照射联合脾-胸腺淋巴细胞输注的方法建立AA小鼠模型。通过血常规检查和骨髓病理学检查，证明AA模型建立成功。流式细胞术检测Th 17和Treg细胞的频率。免疫组化法检测CD 34+、CD 41+细胞，ELISA法检测IL-6、IL-8、IL-17、TNF-α、IFN-γ。结果137 Cs亚致死剂量照射（5戈伊）和脾-胸腺淋巴细胞输注（5 × 10 6）成功建立AA小鼠模型。AA小鼠寿命长，表现为全血细胞减少和骨髓衰竭。AA小鼠Th 17细胞比例明显升高，Treg细胞比例明显降低。AA小鼠骨髓和外周血中IL-6、IL-8、IL-17、TNF-α和IFN-γ水平显著升高，骨髓和外周血中IL-6、IL-8、IL-17、TNF-α和IFN-γ水平显著升高。结论改良的AA小鼠模型符合AA的诊断标准，模拟了人类AA的免疫内环境。AA小鼠模型生存期较长，Th 17/Treg细胞失衡导致CD 34+细胞和CD 41+细胞破坏，是免疫介导的发病机制，适合长期研究。
ABSTRACT Objectives The immune-induced aplastic anemia (AA) mouse model has been used for the study of AA. However, there were no uniform conditions for establishing a model and no assessment of immunological homeostasis. Our study aimed to identify the conditions of establishing a model and assess the AA model in immunology and pathology. Methods We induced an AA mouse model by the combination between sublethal irradiation and spleen-thymus lymphocyte infusion. The success of establishing the AA model was identified by blood routine tests and pathology of bone marrow. The frequency of Th17 and Treg cells was measured by flow cytometry. The frequency of CD34+ and CD41+ cells was detected by immunohistochemical technique.IL-6, IL-8, IL-17, TNF-α and IFN-γ were evaluated by ELISA. Results The 137Cs sublethal irradiation (5 Gy) and spleen-thymus lymphocyte infusion (5 106) induced the AA mouse model successfully. The AA mice had a long lifetime and manifested pancytopenia and bone marrow failure. The percentage of Th17 cells increased and the percentage of Treg cells decreased distinctly in AA mice. The area of hematopoietic tissues and count of CD34+ cells and CD41+ cells were significantly reduced in AA mice.The level of cytokines, IL-6, IL-8, IL-17, TNF-α and IFN-γ, was increased significantly in peripheral blood and bone marrow. Conclusion Our data suggest that the improved AA mouse model conforms to the diagnosis standard of AA and simulates the immune internal environment of human AA. The AA mouse model has a longer lifetime and unbalances of Th17/Treg cells caused the destruction of CD34+ cells and CD41+ cells, which was immune-mediated pathogenesis to adapt to long-term research.