EpCAM蛋白激活DC诱导抗原特异性CTL治疗卵巢癌的实验研究

发布时间：2018-05-13 10:43

本文选题：卵巢癌 + 免疫治疗　；参考：《安徽医科大学》2017年硕士论文

【摘要】：目的探讨Ep CAM蛋白激活树突细胞(DC)诱导产生CD8+细胞毒T淋巴细胞(CD8+CTL)进行卵巢癌免疫治疗的效果,为卵巢癌的临床治疗提供帮助。方法利用Ep CAM蛋白诱导成熟DC同时检测DC表面分子和白介素(IL-10与IL-12)表达量的变化,随后通过Ep CAM-DC诱导Ep CAM抗原特异性CD8+CTL,继而检测Ep CAM-DC-CD8+CTL对正常卵巢上皮细胞IOSE80和卵巢癌细胞SKOV3的杀伤效果,同时检测干扰素(IFN)-γ释放量。随后进一步检测Ep CAM-DC-CD8+CTL对卵巢癌移植裸鼠的肿瘤抑制效果,并通过病理学染色比较治疗前后肿瘤组织变化情况。结果与PBS刺激相比,Ep CAM蛋白能够显著上调DC表面分子CD80、CD83、CD86和HLA-DR水平,依次达到4.79、4.85、4.60和10.91倍;同时Ep CAM蛋白显著提高IL-12释放和显著抑制IL-10分泌(P0.05)。DC-CD8+CTL与Ep CAM-DC-CD8+CTL均引起少量IOSE80细胞凋亡(P0.05),但Ep CAM-DC-CD8+CTL对SKOV3细胞杀伤率是DC-CD8+CTL的6.82倍(P0.05)。动物实验表明,经Ep CAM-DC-CD8+CTL治疗后,BALB/C-nu/nu卵巢癌移植肿瘤体积比明显低于PBS组以及DC-CD8+CTL组,分别达到0.27和0.28倍(P0.05)。HE染色显示Ep CAM-DC-CD8+CTL治疗导致肿瘤组织出现明显的病理学改变。结论Ep CAM蛋白刺激促进了DC成熟继而诱导产生Ep CAM特异性CD8+CTL,Ep CAM-DC-CD8+CTL能够高效的杀伤肿瘤细胞并延迟肿瘤生长,对卵巢癌临床免疫治疗具有重要意义。
[Abstract]:Objective to investigate the effect of immunotherapy of ovarian cancer with Ep CAM protein activated dendritic cell (DC) induced CD8 cytotoxic T lymphocytes (CD8 CTL), and to provide help for clinical treatment of ovarian cancer. Methods mature DCs were induced by EP CAM protein to detect the expression of IL-10 and IL-12 on the surface of DCs at the same time. Then Ep CAM antigen-specific CD8 CTLs were induced by EP CAM-DC, then the killing effect of EP CAM-DC-CD8 CTL on normal ovarian epithelial cells IOSE80 and ovarian cancer cells SKOV3 was detected, and the release of IFN- 纬 from IFN- 纬 was detected. Then the tumor inhibition effect of EP CAM-DC-CD8 CTL on nude mice transplanted with ovarian cancer was further detected, and the changes of tumor tissue before and after treatment were compared by pathological staining. Results compared with PBS stimulation, Ep CAM protein could significantly upregulate the levels of CD80, CD83, CD86 and HLA-DR on DC surface, which were 4.79 ~ 4.85 ~ 4.60 and 10.91 times respectively. At the same time, EP CAM protein significantly increased the release of IL-12 and significantly inhibited the secretion of IL-10 by P0.05N. DC-CD8 CTL and EP CAM-DC-CD8 CTL both induced a small amount of IOSE80 cell apoptosis, but the killing rate of EP CAM-DC-CD8 CTL on SKOV3 cells was 6.82 times as high as that of DC-CD8 CTL. Animal experiments showed that the tumor volume ratio of BALB / C-nunu ovarian cancer transplanted after Ep CAM-DC-CD8 CTL therapy was significantly lower than that of PBS group and DC-CD8 CTL group, and reached 0.27 and 0.28 times P0.05U. He staining showed that EP CAM-DC-CD8 CTL treatment resulted in obvious pathological changes in tumor tissue. Conclusion Ep CAM protein stimulates DC maturation and then induces the production of EP CAM specific CD8 CAM-DC-CD8 CTL, which can effectively kill tumor cells and delay tumor growth. It is of great significance for clinical immunotherapy of ovarian cancer.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.31

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