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目的:建立EB病毒转化B淋巴母细胞系(B-LCL),作为抗原提呈细胞(APC)提呈抗原肽,刺激短期培养的特异性T细胞活化并分泌IFN-γ,从而应用于T细胞表位鉴定中。方法:用B95-8细胞培养上清中的EB病毒转化肾综合征出血热(HFRS)患者PBMC,建立HFRS患者的B-LCL,以自身B-LCL为APC,加载抗原肽后,刺激短期培养的G9L特异的HFRS患者CD8+T细胞系,应用ELISPOT测定CD8+T细胞受到抗原肽刺激后产生IFN-γ的能力。结果:加载过抗原肽G9L或V15R的B-LCL可刺激G9L特异的CD8+T细胞活化并产生IFN-γ,而与G9L无同源序列的I15P则不能刺激G9L特异的CD8+T细胞活化。结论:B-LCL可作为非专职APC有效地将抗原肽提呈给特异性T细胞。
OBJECTIVE: To establish EBV-B lymphoblastoid cell line (B-LCL) as an antigen presenting cell (APC) to present antigenic peptide and to stimulate specific T cell activation and secretion of IFN-γ in short-term culture and to apply it to T cells Epitope identification. Methods: The PBMC of patients with HFRS were transformed by Epstein-Barr virus (EBV) in the culture supernatant of B95-8 cells. B-LCL was established in patients with HFRS. After self-B-LCL was used as APC, Of CD8 + T cell lines of G9L-specific HFRS patients, the ability of CD8 + T cells to produce IFN-γ after antigen peptide stimulation was measured using ELISPOT. Results: B-LCL loaded with antigen peptide G9L or V15R stimulated G9L-specific CD8 + T cell activation and IFN-γ production, whereas I15P, which was not homologous to G9L, stimulated G9L-specific CD8 + T cell activation. Conclusion: B-LCL can be effectively used as a part-time APC to present antigenic peptides to specific T cells.