Yu Dah-Shyong, Lee Chi-Feng, Hsieh Dar-Shih, Chang Sun-Yran
Laboratory of Uro-oncology, Division of Urology, Department of Surgery, Tri-Service General Hospital, Taipei, Taiwan, Republic of China.
Urology. 2004 Mar;63(3):596-601. doi: 10.1016/j.urology.2003.09.039.
To evaluate the antitumor effects of recombinant bacille Calmette-Guérin (BCG) DNA (multi-rBCG) and murine interleukin-12 DNA (mIL-12) vaccines on xenografted MBT-2 murine bladder tumors.
Treatment with combined multi-rBCG and mIL-12 was examined in syngeneic C3H/HeN mice and athymic nude mice. The delivery efficiency of multi-rBCG expression was detected by flow cytometry. Inhibition of tumor growth was monitored, and antitumor effects were evaluated after one dose of electroporation immunogenetherapy, with measurement of cytokines and phenotyping of infiltrating lymphocytes in tumors.
In vivo expression of multi-rBCG was efficient and reached a maximum on day 7 after electroporation. Treatment with multi-rBCG plus mIL-12 significantly inhibited tumor growth in C3H/HeN mice, with increased production of Th1-type cytokines, including interferon-gamma and IL-12. Treatment with multi-rBCG and/or mIL-12 in C3H/HeN mice induced infiltration of CD4+/CD8+ T cells and expansion of natural killer cells within tumors. By contrast, however, athymic nude mice treated in the same way showed no significant immune cells within tumors and died of the fast growing tumors.
Electroporation using multi-rBCG plus mIL-12 could be effective immunotherapy for existing bladder cancer. The antitumor effects correlated with the elicitation of Th1 lymphocytes and natural killer cell-mediated cytotoxic immune responses.
评估重组卡介苗（BCG）DNA（多重组BCG）和小鼠白细胞介素-12 DNA（mIL-12）疫苗对异种移植的MBT-2小鼠膀胱肿瘤的抗肿瘤作用。
在同基因C3H/HeN小鼠和无胸腺裸鼠中检测联合使用多重组BCG和mIL-12的治疗效果。通过流式细胞术检测多重组BCG表达的递送效率。监测肿瘤生长抑制情况，并在一剂电穿孔免疫基因治疗后评估抗肿瘤作用，同时测量细胞因子并对肿瘤中浸润淋巴细胞进行表型分析。
多重组BCG在体内表达有效，电穿孔后第7天达到最大值。多重组BCG加mIL-12治疗显著抑制了C3H/HeN小鼠的肿瘤生长，Th1型细胞因子（包括干扰素-γ和IL-12）的产生增加。在C3H/HeN小鼠中用多重组BCG和/或mIL-12治疗可诱导肿瘤内CD4+/CD8+ T细胞浸润和自然杀伤细胞扩增。然而，相比之下，以同样方式治疗的无胸腺裸鼠肿瘤内未显示出明显的免疫细胞，并死于快速生长的肿瘤。
使用多重组BCG加mIL-12进行电穿孔可能是现有膀胱癌的有效免疫疗法。抗肿瘤作用与Th1淋巴细胞的激发和自然杀伤细胞介导的细胞毒性免疫反应相关。
