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目的:探讨携带癌胚抗原(carcinoembryonic antigen,CEA)基因重组腺相关病毒rAAV/CEA感染直肠癌患者的DC诱导CTL对肿瘤的杀伤作用。方法:抽取直肠癌患者的外周血培养DC细胞,用rAAV/CEA感染DC,DC成熟后与淋巴细胞混合诱导产生CTL,用流式细胞仪分别检测感染和未感染病毒的DC的表型及其诱导的CTL杀伤活性。结果:实验组DC的CD40、CD80、CD86表达显著增高[(81.95±1.45)%vs(72.27±1.59)%,(80.10±1.03)%vs(70.59±1.04)%,(58.93±1.84)%vs(48.66±1.97)%;均P<0.05]。实验组DC中CEA的表达率显著增高[(59.04±1.68)%vs(46.03±1.23)%;均P<0.05]。在效靶比为12.5∶1、25∶1、50∶1、100∶1时,实验组CTL对直肠癌LoVo细胞的杀伤率明显提高[(12.65±0.30)%vs(6.59±0.21)%,(25.77±2.53)%vs(8.35±1.12)%,(43.36±0.83)%vs(12.79±0.23)%,(61.03±1.83)%vs(23.35±0.69)%;均P<0.05],并且随着效应细胞比例的增加,杀伤率也逐渐增加。结论:携带CEA重组腺相关病毒感染DC激活CTL对肿瘤细胞能产生高效的杀伤作用。
OBJECTIVE: To investigate the cytotoxicity of CTL induced by DC induced by recombinant adeno-associated virus (rAAV / CEA) carrying carcinoembryonic antigen (CEA) gene in rectal cancer patients. Methods: DCs from peripheral blood of patients with rectal cancer were cultured, DCs were infected with rAAV / CEA, and then mixed with lymphocytes to induce CTL after maturation. The phenotypes of infected and uninfected DCs were detected by flow cytometry Induced CTL killing activity. Results: The expressions of CD40, CD80 and CD86 in experimental group were significantly higher than those in experimental group [(81.95 ± 1.45)% vs (72.27 ± 1.59)%, (80.10 ± 1.03)% vs (70.59 ± 1.04)%, (58.93 ± 1.84)% vs (48.66 ± 1.97)%, all P <0.05]. The expression of CEA in experimental group was significantly higher than that in control group [(59.04 ± 1.68)% vs (46.03 ± 1.23)%, all P <0.05]. When the target ratio was 12.5:1, 25:1, 50:1, 100:1, the cytotoxicity of CTL on rectal cancer LoVo cells was significantly increased [(12.65 ± 0.30)% vs (6.59 ± 0.21)%, (25.77 ± 2.53)% vs (8.35 ± 1.12)%, (43.36 ± 0.83)% vs (12.79 ± 0.23)%, (61.03 ± 1.83)% vs (23.35 ± 0.69)%, all P <0.05] Increasing the proportion of effector cells, the killing rate is also gradually increased. CONCLUSION: CTLs activated by DCs carrying CEA recombinant adeno-associated virus can efficiently kill tumor cells.