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目的体外建立原发性免疫缺陷病(PIDD)患者永生化B淋巴母细胞系(B-LCL),为进一步开展PIDD相关研究提供资源。方法自PIDD患者外周血中分离单个核细胞,采用EB病毒感染并加入环胞菌素A抑制T淋巴细胞活性的方法建立B-LCL,7 d左右倒置显微镜下观察转化细胞的形态,并进行染色体核型分析。结果转化后的B淋巴母细胞体积增大,增生的淋巴细胞多聚集成团;已建立的永生化细胞能够成功冻存、复苏及传代;转化前后细胞染色体核型无明显改变。结论本实验成功建立了PIDD患者永生化的B-LCL,为进一步开展PIDD的病因学研究及诊断、治疗奠定了基础。
Objective To establish immortalized B lymphoblastoid cell line (B-LCL) in patients with idiopathic immunodeficiency disease (PIDD) in vitro and provide resources for further research on PIDD. Methods Peripheral blood mononuclear cells (PBMCs) were isolated from peripheral blood of patients with PIDD. B-LCL was induced by Epstein-Barr virus (EBV) infection and cyclosporin A-inhibiting T lymphocyte activity. The morphology of transformed cells was observed under inverted microscope for about 7 days, Karyotype analysis. Results The transformed B lymphoblasts increased in size and proliferated lymphocytes aggregated into clusters. The established immortalized cells were successfully cryopreserved, resuscitated and passaged, and the karyotype of the transformed cells did not change significantly. Conclusion This study successfully established immortalized B-LCL in patients with PIDD, which lays the foundation for the further study of etiology and diagnosis and treatment of PIDD.