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目的 观察树突细胞免疫对 HPV感染阳性角朊细胞 (HIPK)增殖特性的影响及诱发杀伤性 T淋巴细胞 (CTL)细胞毒活性的变化 .方法 用计数法绘制细胞生长曲线、流式细胞仪 (FCM) )检测 DNA含量及细胞周期的变化 ,以观察HIPK增殖特性变化 .同时用 5 1 Cr释放法测定树突细胞免疫前后细胞毒性淋巴细胞 (CTL )对 HIPK的细胞毒作用 .结果 树突细胞免疫后 ,HIPK生长延缓 ,增殖幅度降低 ,倍增时间 (Dt)延长 ,HIPK的 Dt为 2 4.7h,DCs细胞培养上清作用后 HIPK的 Dt为 41.6 h. FCM检测 HIPK的 G0 / G1,S,G2 / M各时相细胞百分比 (% )分别为 :5 7.6 ,2 8.5 ,13.9;DCs细胞培养上清作用组分别为 6 1.7,2 0 .3,11.2 .两者增殖指数(PI)比较差异显著 (P<0 .0 1) .另外没有观察到 HIPK凋亡 ,而 DCs培养上清作用组却观察 6 .8%的凋亡率 .5 1 Cr释放实验中 ,效 -靶比为 10∶ 1,5∶ 1,2∶ 1时 ,DCs治疗组 CTL对HIPK杀伤率 (% )分别为 37.6 ,2 1.4,13.7;而对照组为 5 .6 ,2 .9,1.6 .结论 树突细胞免疫后 HIPK出现了一定的增殖抑制现象 ,同时诱导了 CTL的杀伤作用 .
Objective To observe the effect of dendritic cell immunity on the proliferation of HPV-positive keratinocytes (HIPK) and the cytotoxicity of cytotoxic T lymphocytes (CTL) .Methods The cell growth curve was drawn by flow cytometry FCM) were used to detect the changes of DNA content and cell cycle in order to observe the proliferation characteristics of HIPK.Cell cytotoxicity of cytotoxic lymphocytes (CTL) before and after dendritic cell immunization with HIPK was measured by 5 1 Cr release assay.Results Dendritic cells After immunization, the growth of HIPK was delayed, the proliferation amplitude was decreased, the doubling time (Dt) was prolonged, the Dt of HIPK was 2 4.7 h, and the Dt of HIPK was 41.6 h after DCs cultured in supernatant of DCs.GM / G1, The percentages of cells in G2 / M phase were (5 7.6), (2 8.5) and (13.9) respectively, and the proliferative index (PI) (P <0.01) .In addition, no apoptosis of HIPK was observed, while the apoptotic rate of 6.8% in DCs cultured supernatant group was observed.When the 5 1 Cr release experiment was carried out, the efficiency-target ratio was 10: 1, 2: 1, the killing rate (%) of CTL to HIPK in DCs treatment group was 37.6, 2 1.4,13.7, while the control group was 5.6, 2.9, 1.6 Conclusion HIPK dendritic cells after a certain degree of inhibition of proliferation, while inducing CTL killing effect.