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冻融肿瘤疫苗的制备及对小鼠恶性黑色素瘤抑瘤作用的实验研究
Preparation of Frozen Tumor Vaccines and Their Antitumor Effect Study to Malignant Melanoma Carcinoma of Mice
【作者】 刘佳；
【导师】 曹培国；
【作者基本信息】 中南大学 ， 肿瘤学， 2010， 硕士
【摘要】 目的：探讨冻融肿瘤疫苗对C57BL/6J小鼠黑色素瘤的抑制作用。方法：(1)将B16Fo恶性黑色素瘤细胞皮下接种C57BL/6J小鼠建立肿瘤模型后,行氩氦刀冷冻消融肿瘤组织。(2)取冻融肿瘤组织联合弗氏不完全免疫佐剂制成肿瘤疫苗。(3)12只C57BL/6J小鼠随机分为实验组和对照组,每组6只。实验组小鼠注射肿瘤疫苗,每周1次,共4次,并于每次注射疫苗后眼眶采血,测量外周血CD4+、CD8+免疫指标变化；对照组小鼠注射佐剂和生理盐水。第四次疫苗注射完成后各组小鼠均皮下接种黑色素瘤细胞,观察各组小鼠的成瘤率、出瘤时间,肿瘤生长状态以及生存期。结果：1.实验组及对照组小鼠全部成瘤,成瘤率为100%。实验组和对照组小鼠平均出瘤时间分别为11.35±1.6天和6.7±1.2天,两组比较差异有显著性(P＜0.01)；平均生存时间分别为36.0±2.8天和25.6±1.5天,两组比较差异有显著性(P＜0.01)。2.实验组小鼠外周血T细胞亚群在免疫后呈动态变化,CD4+、CD4+／CD8+比值在免疫后逐渐增加,在第2周时与疫苗接种前相比差异有显著性(P＜0.01),在第4周时达到高峰；CDs+在免疫后开始下降,在第4周时与疫苗接种前相比差异有显著性(P＜0.01)。结论：1.冻融肿瘤疫苗能有效延缓小鼠恶性黑色素瘤的生长,延长荷瘤小鼠的生存期。2.冻融肿瘤疫苗能刺激机体细胞免疫,改善机体的免疫状况。
【Abstract】 Objective:To investigate the inhibitory effect of tumor vaccines(the necrotic tumor tissues after cryosurgery) in mice with malignant melanoma carcinoma.Methods:(1)The tumor model was established by subcutaneous implantation of B16Fo cells in C57BL/6J mice,then treated with cryosurgery.(2)Frozen tumor tissues combined with incomplete Freund adjuvant were made into tumor Vaccines.(3)Twelve C57BL/6J mice were randomly divided into 2 groups:experimental group and control group.Experimental mice group was injected with tumor Vaccines and blood was taken from the orbit after each vaccination,then the changes of CD4+、CD8+ were analyzed. Control mice group was injected with Adjuvant and NS.B16Fo cells was implanted subcutaneously into mice after 4th vaccination. The tumor growth and survival period were observed.Results:1.The tumor formation rate of each group was 100%. The mean tumor growth time of experimental and control group were 11.3±1.6 and 6.7d±1.2 days,respectively(P<0.01),and the mean survival time of two group mice were 36.0±2.8 and 25.6d±1.5 days,respectively (P<0.01).2.1n experimental group, T cell subsets in peripheral blood after immunization showed dynamic changes.CD4+、CD47/CD8+ gradually increased and CD8+ decreased after immunization,and reached a peak value in the 4th week(P<0.01).Conclutions:1.Tumor vaccine can effectively delay the growth of malignant melanoma carcinoma and prolong survival time of immunized mices.2.Tumor vaccine can stimulate body cell immunity and improve immune function.