前列腺癌PSA特异性树突状细胞瘤苗的构建及其体外免疫活性的观察
Journal Title: Di-er Junyi Daxue Xuebao - Year 2009, Vol 30, Issue 3
Abstract
目的:构建前列腺癌PSA特异性树突状细胞(PSADC)瘤苗,并观察其体外免疫活性,为后续研究奠定基础。方法:分离骨髓前体细胞,大量制备骨髓DCs(bone marrow derived DC,BMDC);分别以PSA、癌细胞裂解产物(lysate,Lys)、无关蛋白卵清蛋白(Ova)冲击DC制备PSADC、LysDC、OvaDC瘤苗。ELISA法检测PSADC瘤苗培养上清中细胞因子(IL12 p70和IL1β)水平的变化;观察PSADC瘤苗刺激抗原特异性T细胞增殖活性和诱导抗原特异性CTL杀伤活性,并与LysDC、OvaDC瘤苗及未加入抗原冲击的DC(NonDC)组作比较。结果:成功获得成熟DC,纯度可以达到>95%。ELISA法检测结果表明,PSADC、LysDC和OvaDC组培养上清中IL12 p70和IL1β水平均较NonDC组明显升高(P<0.05)。混合淋巴细胞培养(MLR)显示:PSADC、LysDC组DCs刺激CD4+T细胞增殖的能力明显优于OvaDC、NonDC组(P<0.01);PSADC、LysDC组培养上清中IL2、IFNγ水平明显高于后两者(P<0.01),而IL10和IL4水平下调(P<0.05)。PSADC、LysDC组可产生针对PSA和肿瘤裂解物的DTH反应,而对无关抗原无效(P<0.05)。与OVADC、NonDC组相比,LysDC、PSADC组体外诱导的CTL细胞对LNCaP细胞的杀伤活性较强 (P<0.05),且具有抗原特异性。结论:利用PSA蛋白冲击DC可成功制备PSA特异性DC瘤苗,该瘤苗体外具有较强的免疫活性,能有效杀伤LNCaP细胞。
Authors and Affiliations
Dan feng XU, Yi GAO, Yu shan LIU, Xin gang CUI, Jian ping CHE, Lei YIN, Ji zhang XING, Ya cheng YAO, Ji zhong REN, Zhi lian MIN
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