【摘要】 目的:探讨人白细胞介素18在鼻咽癌细胞中的表达能否提高鼻咽癌病人外周血CD8+T细胞的杀伤活性以及作用途径。方法:构建人白细胞介素18的真核表达载体pcDNA3．1(-)/HIL-18,转染人鼻咽癌细胞株SUNE;以转染的SUNE细胞和未转染的 SUNE细胞为靶细胞,以鼻咽癌患者外周血的 CD8+T细胞为效应细胞,混合培养 12h,用 LDH释放法测定 CD8+T细胞的细胞毒活性;用免疫组化法测定混合培养物中 CD8+T细胞上 Perforin、Fas－L、Granzyme B的表达。结果:所构建的真核表达载体在鼻咽癌细胞中能高效表达hIL-18;ELISA法测得转染阳性细胞培养上清液中hIL-18的含量为(85±10)pg/ml,而未转染的SUNE细胞的培养上清液中hIL-18的含量低于5pg/ml。将表达hIL－18的SUNE细胞与鼻咽癌患者外周血CD8+T细胞混合培养12h后,CD8+T细胞的细胞毒活性显著增强(P＜0．111),尤其是当效应细胞/靶细胞为10:1时,CD8+细胞对转染的SUNE细胞的裂解率高达46．7%,而CD8+T细胞对未转染的SUME细胞的裂解率仅为4．6%。免疫组化法表明,这种杀伤活性的增强与CD8+T细胞表达Perforin有关,而与Fas-L和Granzyme B途径无关。结论:hIL-18能显著增强鼻咽癌患者外周血口CD8+T细胞的体外杀伤活性,这种杀伤活性与 CDS8+细胞表达Perforin有关。更多还原

【Abstract】 The study is designed to investigate if human interleukin 18 (hIL-18) cDNA expressed in nasopharyngeal carcinoma (NPC) cells could enhance the cytotoxicity of CD8+ T cells from the PBMC of the MFC-bearing patient and how to enhance it.Methods: A constructed eukaryotic expression vector for human IL-18 ,pcDNA3.1(-)/hIL-18, was used to transfect the NPC cell strain-SUNE cells. Then, the transfected SUNE cells were mix-cultured with CD8+ T cells which were separated from the PBMC of the patient with NPC, the cytotoxicity was measured by lactate dehydrogenase(LDH) releasing method, the expression for Perforin, Fas-L and Granzyme B were detected with the mix-cultures. Results: The eukaryotic expression vector could highly express hIL-18 in transfected SUNE cells and only traces of the protein exist in non-transfected SUNE cells. The concentration of hIL-18 in supernatant of the transfected SUNE cells was (85 ± 10) pg/ml, but less than 5 pg/ml of hIL-18 contained in supernatant of the non-transfected SUNE cells.The expression of hIL-18 in SUNE cells could enhance the cytotoxicity of CD8+ T cells from the NPC patient significantly( P < 0.001), and this activity was related to the expression of perforin on CD8+ T cells. Conclusion:hIL- 18 can increase the cytotoxicity of CD8+ T cells in vitro and induce the expression of perforin on CDS+ T cells.更多还原