Activation and dramatically increased cytolytic activity of tumor specific T lymphocytes after radio-frequency ablation in patients with hepatocellular carcinoma and colorectal liver metastases

瞄准：为了如果特定的细胞毒素的 T 房间回答能与恶意的肝在病人被导致，估计，肿瘤与无线电频率对待脱离(RFA ) 。方法：有颜色的肝转移的六个病人表面的癌症并且 6 与肝细胞，癌(HCC ) 经历了 RFA。血以前被取样，在 RFA 以后的 4 和 8 wk。测试抗原是活体检视从每个病人获得的自体同源的肝和肿瘤溶解产物。外部 T 房间激活被一个干扰素鲸鱼群妈(IFNgamma ) 估计分泌物试金和流动血细胞计数。T 房间是双 stained 让 CD4/CD8 和 IFNgamma 检测细胞毒素的 T 房间。积极的 IFNgamma 和 IFNgamma 否定 T 房间的比率作为刺激索引(SI ) 被决定。估计细胞溶解的活动， T 房间与人的 CaCo 颜色被共同孵化 cytosolic adenylate 激酶的表面的癌症和 HepG2 HCC 房间和版本被酶试金测量。结果：在 RFA SI 前是 0.021 (+/- 0.006 ) 为 CD4 (+) 并且 0.022 (+/- 0.004 ) 为对非恶意的肝织物的 CD8 (+) T 房间并且 0.018 (+/- 0.005 ) 为 CD4 (+) 并且 0.021 (+/- 0.004 ) 为对自体同源的肿瘤织物的 CD8 (+) 房间。在对肿瘤织物的 RFA SI 以后的四个星期增加了到 0.109 (+/- 0.005 )为 CD4 (+)并且 0.11 (+/- 0.012 )为对 HCC 的 CD8 (+) T 房间，并且到 0.115 (+/- 0.031 )为 CD4 (+)并且 0.15 (+/- 0.02 )为为颜色的 CD8 (+)房间表面的转移( P 【 0.0001 )。没有增加的 SI 被观察与非，恶性瘤织物根本预定点。与 2.62 在对各自的癌症房间的 RFA 细胞溶解的的活动前是低的(+/- 0.37 ) 相对的光单位(RLU ) ，而是超过 100 合拢的玫瑰在 RFA 以后的 4 和 8 wk。自发的版本是 2% 最大值在所有实验释放的 【 。结论：病人与主要并且肝的继发性瘤在 RFA 以后与 CD8 (+) T 房间的一项戏剧性地增加的肿瘤 specific 细胞溶解的活动显示出重要肿瘤特定的细胞毒素的 T 房间刺激。

AIM： To assess if a specific cytotoxic T cell response can be induced in patients with malignant liver tumors treated with radio-frequency ablation （RFA）. METHODS： Six Patients with liver metastases of colorectal cancer and 6 with hepatocellular carcinoma （HCC） underwent RFA. Blood was sampled before, 4 and 8 wk after RFA. Test antigens were autologous liver and tumor lysate obtained from each patient by biopsy. Peripheral T cell activation was assessed by an interferon gamma （IFNγ） secretion assay and flow cytometry. T cells were double-stained for CD4/CD8 and IFNγ to detect cytotoxic T cells. The ratio of IFNγ positive and IFNγ negative T cells was determined as the stimulation index （SI）. To assess cytolytic activity, T cells were co-incubated with human CaCo colorectal cancer and HepG2 HCC cells and release of cytosolic adenylate kinase was measured by a luciferase assay. RESULTS： Before RFA SI was 0.021 （±0.006） for CD4^＋ and 0.022 （± 0.004） for CD8^＋T cells against nonmalignant liver tissue and 0.018 （± 0.005） for CD4^＋ and 0.021 （± 0.004） for CD8^＋ cells against autologous tumor tissue. Four weeks after RFA SI against tumor tissue increased to 0.109 （± 0.005） for CD4＋ and 0.11 （± 0.012） for CD8＋ T cells against HCC, and to 0.115 （± 0.031） for CD4^＋ and 0.15 （± 0.02） for CD8^＋ cells for colorectal metastases （P 〈 0.0001）. No increased SI was observed with nonmalignant tumor tissue at all time points. Before RFA cytolytic activity against the respect（ve cancer cells was low with 2.62 （± 0.37） relative luminescence units （RLU）, but rose more than 100 fold 4 and 8 wk after RFA. Spontaneous release was 〈 2% of maximum release in all experiments. CONCLUSION： Patients with primary and secondary tumors of the liver show a significant tumor-specific cytotoxic T-cell stimulation with a dramatically increased tumor specific cytolytic activity of CD8^＋ T cells after RFA.