树突细胞免疫对HPV感染阳性角朊细胞生物学特性影响

Effect of dendritic cell immunization on biological characteristics of HPV infected keratinocytes

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 .

AIM To investigate the characteristic change of HPV infection positive keratinocytes (HIPK)in culture after dendritic cell immunization. METHODS By comparing the difference of the growth curve with cytometre assay, DNA content and cell cycle of HIPK with flow cytometre(FCM) assay, we studied the growth characteristic distinctness of HIPK cultured with the supernatant of dendritic cell and the cytotoxicity alteration of the cytotoxic lymphocyte(CTL) to HIPK was evaluated with 51 Cr release assay. RESULTS After dendritic cell immunization, the HIPK proliferation was lagged, The double proliferation time(Dt) of HIPK was 24.7 h, and the Dt of HIPK treated with supernatant of DCs was 41.6 h. With FCM assay, the percentage (%) of the cells at distinct stage G0/G1, S, G2/M phase of HIPK was 57.6,28 5,13.9 while the percentage of HIPK cultured with the supertant of DCs was 61.7,20.3,11.2 respectively. The proliferation indexes of them were significantly different ( P <0 01, χ 2 test).When effect/target ratios were 10:1,5:1,2:1, the cytotoxicity rates( 51 Cr release rate/%) of DCs treated group were 37.6,21.4,13.7 respectively, while those of control group were 5.6,2.9,1.6 respectively with 51 Cr release assay. The significant differences between them ( P <0 01) were evaluated by χ 2 test. The apoptosis was found only in HIPK treated with supernatant of DCs, whose apoptotic percentage was 6.8% assayed with FCM. CONCLUSION Dendritic cell immunization can restrain proliferation of HIPK as well as killing it by activated CTL.