McKinnon J G, Lalani A, Sy L, Bear H D
Tom Baker Cancer Center, University of Calgary, Alberta, Canada.
Surgery. 1993 May;113(5):536-40.
Cytotoxic T lymphocytes (CTL) have been shown to be useful for adoptive immunotherapy in malignancy. Traditional sources for CTL, such as tumor-infiltrating lymphocytes, have limitations. It would therefore be useful to develop a method of generating antitumor CTL from a renewable source such as peripheral blood.
DBA/2 mice were injected intradermally in the abdominal wall with the murine tumor PHS-5 and killed 14 days later. Peripheral blood lymphocytes were harvested and cultured with 20 units/ml interleukin-2 and autologous tumor-stimulator cells treated with mitomycin C. Cultures were split when greater than 2 x 10(6) cells/well, fed every 3 days and stimulated weekly.
Lymphocytes expanded greater than 130,000-fold during 8 weeks. Specific cytotoxicity was shown with 51Cr release assay. Withdrawal of repeated stimulation with autologous tumor resulted in failure of cells to expand in culture and loss of cytotoxicity. In vivo administration showed marked reduction of 10-day liver metastases, indicating therapeutic efficacy.
These data demonstrate a successful animal model of adoptive immunotherapy with CTL generated from peripheral blood lymphocytes.
细胞毒性T淋巴细胞（CTL）已被证明在恶性肿瘤的过继性免疫治疗中有用。传统的CTL来源，如肿瘤浸润淋巴细胞，存在局限性。因此，开发一种从外周血等可再生来源产生抗肿瘤CTL的方法将是有用的。
将小鼠肿瘤PHS-5皮内注射到DBA/2小鼠的腹壁，14天后处死小鼠。收集外周血淋巴细胞，与20单位/毫升白细胞介素-2和经丝裂霉素C处理的自体肿瘤刺激细胞一起培养。当细胞密度大于2×10⁶个细胞/孔时进行传代培养，每3天换液一次，每周进行一次刺激。
淋巴细胞在8周内扩增超过130000倍。通过⁵¹Cr释放试验显示出特异性细胞毒性。停止用自体肿瘤反复刺激导致细胞在培养中无法扩增并丧失细胞毒性。体内给药显示10天肝转移明显减少，表明具有治疗效果。
这些数据证明了一种成功的过继性免疫治疗动物模型，该模型使用从外周血淋巴细胞产生的CTL。
