埃泼斯坦-巴尔病毒潜伏膜蛋白2A特异性细胞毒性T淋巴细胞的体外诱导研究

Induction of Epstein-Barr Virus Latent Membrane Protein 2A-Specific Cytotoxic T Lymphocytes in vitro

目的:利用含埃泼斯坦-巴尔病毒(Epstein-Barr virus,EBV)潜伏膜蛋白2A重组痘苗病毒(rVV-LMP2A)感染的人树突状细胞(DC)体外诱导EBV-LMP2A特异性细胞毒性T淋巴细胞(CTL)。方法:rVV-LMP2A感染的DC分别在培养第1、8天刺激相同MHC背景的人外周血单个核细胞(PBMC),在IL-2作用下体外诱导EBV特异性CTL。培养第15天乳酸脱氢酶法检测CTL的特异性杀伤活性。结果:诱导的CTL对rVV-LMP2A感染的靶细胞在效靶比为5:1、10:1和15:1时,特异性杀伤率分别为63.5％、65.5％和67.9％。结论:rVV-LMP2A感染的DC刺激相同MHC背景的PBMC可诱导出较高杀伤活性的EBV特异性CTL。

Objective: To induce the specific cytotoxic T lymphocytes (CTL) in vitro using human monocyte-derived dendritic cells (DC) infected with recombinant vaccinia virus carrying Epstein-Barr virus latent membrane protein 2A (EBV-LMP2A) gene (rVV-LMP2A). Methods: With the treatment of IL-2, EBV-specific CTL was induced by coculture of rVV-LMP2A-infecting DC with the PBMC of identical donor. Specific cytotoxic competence of the CTL was measured by using lactate dehydrogenase (LDH) release assay. Results: The specific cytotoxicity of the CTL with the effect: target (E= T) ratio of 5- 1, 10: 1 and 15: 1 was 63. 5%, 65. 5% and 67. 9% respectively. Conclusion: The EBV-specific CTL can be efficiently induced by DC infected with rVV-LMP2A in vitro.