ALDH1A1 mRNA负载树突状细胞疫苗体外诱导抗鼻咽癌干细胞免疫反应OA
The specific immune response of dendritic cell vaccine pulsed with ALDH1A1 mRNA on nasopharyngeal carcinoma stem cells in vitro
目的:探讨乙醛脱氢酶1A1(ALDH1A1)mRNA负载树突状细胞(DC)疫苗体外诱导抗鼻咽癌干细胞免疫反应的能力.方法:体外转录ALDH1A1 mRNA并电转染至人外周血DC,构建ALDH1A1-DC疫苗,转染EGFP mRNA为对照.流式细胞术检测DC表型、淋巴细胞亚群、细胞毒性T 淋巴细胞(CTL)增殖和细胞因子分泌.分选ALDH+和ALDH-C666-1细胞,检测干细胞标志物(CD24、CD133)和干性基因(OCT4、SOX2、Nanog)的表达水平,采用CCK-8法评估ALDH1A1-DC疫苗诱导的特异性免疫杀伤能力.结果:EGFP-DC荧光表达率显著高于未转染DC(P<0.000 1),ALDH1A1-DC抗原提呈分子HLA-ABC、共刺激分子CD86、CD40和CCR7表达与未转染DC相比显著增强(P=0.001 3、P=0.005 9、P=0.000 4及P=0.001 9).与单独淋巴细胞组(T)及空载DC诱导的淋巴细胞组(DC-T)相比,ALDH1A1-DC诱导的CTL比例最高(均P<0.000 1),分裂指数最高(P=0.000 2、P=0.000 3),且CD8+CD69+(P=0.000 5、P<0.000 1)和CD8+IFN-γ+(P=0.012 6、P=0.001 9)细胞亚群比例均显著提高.ALDH1A1-DC-T细胞对ALDH+C666-1悬浮细胞球的杀伤效果显著优于ALDH-细胞球(P=0.001 0),且与其他各组淋巴细胞相比表现出较强的抗ALDH+C666-1悬浮细胞球的免疫效应(P=0.001 0,P<0.000 1).然而,HLA-ABC阻断后其对ALDH+C666-1细胞的特异性杀伤效应显著降低(P<0.000 1).结论:mRNA转染DC后可翻译蛋白并促进DC成熟,增强其诱导特异性CTL免疫反应的能力.
Objective:To investigate the ability of dendritic cell(DC)vaccine loaded with aldehyde dehydrogenase 1A1(ALDH1A1)mRNA to enhance anti-nasopharyngeal carcinoma stem cell immune responses in vitro.Methods:ALDH1A1 mRNA was transcribed in vitro and electroporated into human peripheral blood-derived DCs to construct ALDH1A1-DC vaccine;EGFP mRNA was used as a transfection control.Flow cytometry was employed to analyze DC phenotypes,lymphocyte subsets,cytotoxic T lymphocyte(CTL)proliferation,and cytokine secretion.ALDH+and ALDH-C666-1 cells were sorted;expression of stem cell markers(CD24,CD133)and stemness-associated genes(OCT4,SOX2,Nanog)was analyzed.The specific cytotoxic capacity induced by ALDH1A1-DC was evaluated using the Cell Counting Kit-8(CCK-8)assay.Results:EGFP-DC exhibited significantly higher fluorescence expression than untransfected DCs(P<0.000 1).Moreover,ALDH1A1-DC significantly enhanced the expression of antigen-presenting molecules HLA-ABC,co-stimulatory molecules CD86,CD40,and CCR7 compared to untransfected DCs(P=0.001 3,P=0.005 9,P=0.000 4,P=0.001 9).Compared with the T lymphocyte group(T)and the DC-T group(DC-induced T lymphocytes),ALDH1A1-DC-induced T lymphocytes(ALDH1A1-DC-T)exhibited the highest proportion of CTL(P<0.000 1),the highest proliferation index(P=0.000 2 and P=0.000 3,respectively),the highest proportions of CD8+CD69+cells(P=0.000 5 and P<0.000 1,respectively),and the highest proportions of CD8+IFN-γ+subsets(P=0.012 6 and P=0.001 9,respectively).CCK-8 assay demonstrated that ALDH1A1-DC-T exhibited significantly enhanced cytotoxicity against ALDH+C666-1 suspension spheres compared to ALDH-spheres(P=0.001 0).Furthermore,ALDH1A1-DC-T elicited the most potent anti-ALDH+C666-1 immune response among all lymphocyte groups tested(P=0.001 0 and P<0.000 1,respectively).However,HLA-ABC blockade significantly attenuated this specific cytotoxicity(P<0.000 1).Conclusion:mRNA transfected into DCs can be translated into protein and promote the maturation of DCs and enhance the ability of DCs to induce specific CTL immune response.
谢云青;许扬梅;陈珊珊;林晓为;刘沁颖
福建省肿瘤医院暨福建医科大学肿瘤临床医学院 福建省肿瘤生物治疗重点实验室,福建 福州 350014福建省肿瘤医院暨福建医科大学肿瘤临床医学院 福建省肿瘤生物治疗重点实验室,福建 福州 350014福建省肿瘤医院暨福建医科大学肿瘤临床医学院 福建省肿瘤生物治疗重点实验室,福建 福州 350014福建省肿瘤医院暨福建医科大学肿瘤临床医学院 福建省肿瘤生物治疗重点实验室,福建 福州 350014福建省肿瘤医院暨福建医科大学肿瘤临床医学院 福建省肿瘤生物治疗重点实验室,福建 福州 350014
医药卫生
肿瘤干细胞乙醛脱氢酶1A1树突状细胞细胞毒性T淋巴细胞鼻咽癌
cancer stem cell(CSC)aldehyde dehydrogenase 1A1(ALDH1A1)dendritic cell(DC)cytotoxic T lymphocyte(CTL)nasopharyngeal carcinoma
《中国肿瘤生物治疗杂志》 2026 (5)
486-493,8
福建省卫生健康委员会医学创新课题(2023CXA036)福建省科技创新联合资金项目(2024Y9622)
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