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目的:研究肿瘤患者树突状细胞经MAGE-3/CEA(HLA-A2/A24+)肽疫苗冲击后诱导特异性CTL的杀伤活性。方法:选择MAGE-3-HLA-A2/A24+或CEA-HLA-A24肿瘤患者,收集PBMNC中的贴壁细胞在含有rhGM-CSF、rhIL-4的1640中培养诱导DC细胞,第7日加入HLA-A2-MAGE-3,HLA-A24-MAGE-3、HLA-A24-CEA肽冲击。以肽冲击后的DC与未经纯化的T细胞混合培养诱导的CTL作为效应细胞,Mel526,803,Raji,K562作为靶细胞,以LDH法检测特异性CTL的杀伤力。结果:未纯化的T细胞及肽冲击后的DC与该T细胞混合培养后的细胞表型变化表明CD3、CD4、CD25、CD16/CD56均无显著性差异,表达的CD8、CD86、CD69、CD45RO/CD8、HLA-DR有显著性差异。T-IL-2细胞与T-DC-P细胞对Raji和K562细胞株的杀伤活性无显著性差异。T-IL-2细胞与T-DC-P细胞对Mel526和803细胞株的杀伤活性有显著性差异。结论:肽疫苗可以诱导MAGE、CEA特异性的CTL应答。DC为基础的疫苗是肿瘤免疫治疗的重要的新方法,因其简便、快速的特点,具有良好的前景。
OBJECTIVE: To study the killing activity of dendritic cells (CTLs) induced by dendritic cells of MAGE-3 / CEA (HLA-A2 / A24 +) peptide vaccine in tumor patients. Methods: MAGE-3-HLA-A2 / A24 + or CEA-HLA-A24 tumor patients were selected. Adherent cells from PBMNC were collected to induce DCs in 1640 cells containing rhGM-CSF and rhIL-4. -A2-MAGE-3, HLA-A24-MAGE-3, HLA-A24-CEA peptide. The specific CTLs were detected by LDH method using CTLs induced by peptide-challenged DC mixed with non-purified T cells as effector cells, Mel526, 803, Raji and K562 as target cells. Results: The phenotypic changes after unincubated T cells and peptide-challenged DCs mixed with the T cells showed that CD3, CD4, CD25 and CD16 / CD56 showed no significant difference. The expression of CD8, CD86, CD69 and CD45RO / CD8, HLA-DR were significantly different. T-IL-2 cells and T-DC-P cells on Raji and K562 cell lines showed no significant difference in cytotoxic activity. The killing activity of T-IL-2 cells and T-DC-P cells on Mel526 and 803 cell lines were significantly different. Conclusion: Peptide vaccine can induce MAGE and CEA specific CTL responses. DC-based vaccine is an important new method of tumor immunotherapy, because of its simple, rapid characteristics, has good prospects.