摘要
目的研究在体外负载前列腺干细胞抗原(PSCA)制备树突状细胞(DC)疫苗的方法。方法应用PSCA融合蛋白冲击致敏由外周血单个核细胞分离培养得到的人DC,采用流式细胞仪检测其活化前、后的表型变化,混合淋巴细胞反应检测其在体外刺激淋巴细胞增殖的能力。结果DC-PSCA组白细胞介素(IL)-12(p40)和IL-1β浓度分别为(183.0±5.0)和(1183.0±30.9)ng/L,显著高于DC-TRX组[(162.0±2.5)和(971.0±10.4)ng/L,P值均<0.01]和DC组[(30.0±2.6)和(204.0±6.1)ng/L,P值均<0.01]。不同抗原冲击活化后,DC-PSCA组人类白细胞抗原(HLA)-DR、CD80、CD86、CD40的荧光强度(除去同型对照抗体荧光强度后的数值)分别为101.7±9.8、334.2±18.3、371.2±22.5和102.3±8.2,显著高于DC-TRX组(88.4±6.3、304.3±19.4、315.5±24.7和94.6±7.9,P值均<0.05)和DC组(68.8±3.0、283.2±23.1、265.4±18.8和77.2±4.6,P值均<0.05)。经肿瘤坏死因子-α活化后的DC-PSCA组放射性活度值(cpm值)显著高于其他两组(P值均<0.05)。结论PSCA融合蛋白体外冲击DC是一种制备DC疫苗的有效方法。
Objective To establish a method for preparing dendritic cell （DC） vaccine against prostate cancer by loading the DCs with prostate stem cell antigen （PSCA） in vitro, Methods The DCs were isolated from human peripheral blood mononuclear cells（PBMCs） and the DC vaccines generated by PSCA pulsing. The phenotype of the antigen-loaded DCs was assayed by flow cytometry before and after activation, and their capacity to activate T lymphocytes assessed by mixed lymphocyte reaction（MLR）, Results The concentrations of IL-12 （[183. 0 ± 5.0] ng/L） and IL-1β （[1183.0 ± 30. 9] ng/L） in DC-PSCA group were significantly higher than those in DC-TRX group （[162.0 ± 2.5] and [971.0 ± 10.4] ng/L） and DC group （[30.0 ± 2.6] and [204.0 ± 6.1] ng/L, all P 〈 0.01）. After pulsed activation, the fluorescence densities（after adjusting the value of control antibody of the same type） of HLA-DR, CDS0, CD86, and CD40 DC in DC-PSCA group were 101.7 ± 9.8, 334.2 ± 18.3, 371.2 ± 22.5 and 102.3 ± 8.2, respectively, all higher than those in the DC-TRX group （68.8 ± 3.0, 283. 2 ± 23. 1, 265.4±18.8 and 77.2 ± 4.6, respectively） and DC group（68.8 ± 3.0, 283.2±23.1, 265.4 ± 18.8 and 77.2 ± 4.6, respectively, all P 〈 0.05）. The cpm value in DC-PSCA group was significantly higher than those in the other two groups after activation of TNF-α（both P 〈 0.05）. Conclusion DC pulsing with PSCA fusion protein is an effective strategy for generating DC vaccines against prostate cancer in vitro. （Shanghai Med J, 2007, 30 ： 924-927）
出处
《上海医学》
CAS
CSCD
北大核心
2007年第12期924-927,共4页
Shanghai Medical Journal
基金
国家自然科学基金(30400445)
上海市卫生局科技发展基金(034117)资助项目
