Human cytokine-induced killer cells have enhanced in vitro cytolytic activity via non-viral interleukin-2 gene transfer.
Human cytokine-induced killer cells have enhanced in vitro cytolytic activity via non-viral interleukin-2 gene transfer.
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DOI：
10.1186/1479-0556-2-12
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发表时间：
2004
期刊：
影响因子：
--
通讯作者：
Schmidt-Wolf IG
中科院分区：
文献类型：
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作者：
Nagaraj S;Ziske C;Schmidt-Wolf IG
通过基因修饰的免疫效应细胞调节免疫系统在恶性肿瘤的治疗中具有潜在的治疗价值。白细胞介素-2 （IL-2）是一种重要的细胞因子，可诱导有效的抗肿瘤反应。细胞因子诱导杀伤细胞（CIK）是一种高效的细胞毒性效应细胞，能够裂解肿瘤细胞靶点，并能够以非mhc限制的方式识别这些细胞。树突状细胞（DC）是主要的抗原呈递细胞。本研究评价了用IL-2非病毒转染CIK细胞，并与脉冲和非脉冲DC共培养CIK细胞的抗肿瘤作用。用编码人IL-2的质粒转染人外周血CIK细胞。转染包括电参数和特定溶液的组合，通过使用nucleoector®电穿孔系统将质粒直接传递到细胞核。与模拟转染CIK细胞（31 pg/106细胞）相比，核转染导致IL-2的产生平均为478.5 pg/106细胞（范围为107.6-1079.3 pg/106细胞/24 h） （P = 0.05）。在与DC共培养后，通过细胞毒性试验评估其体外功能能力。与未转染DC的CIK细胞（36.5±5.3%）相比，转染DC的CIK细胞对人胰腺癌细胞株Dan G细胞的裂解活性显著提高（58.5±3.2%）（P = 0.03）。
Modulation of the immune system by genetically modified immunological effector cells is of potential therapeutic value in the treatment of malignancies. Interleukin-2 (IL-2) is a crucial cytokine which induces potent antitumor response. Cytokine-induced killer cells (CIK) have been described as highly efficient cytotoxic effector cells capable of lysing tumor cell targets and are capable of recognizing these cells in a non-MHC restricted fashion. Dendritic cells (DC) are the major antigen presenting cells. This study evaluated the antitumor effect of CIK cells which were non-virally transfected with IL-2 and co-cultured with pulsed and unpulsed DC. Human CIK cells generated from peripheral blood were transfected in vitro with plasmid encoding for the human IL-2. Transfection involved a combination of electrical parameters and a specific solution to deliver plasmid directly to the cell nucleus by using the Nucleofector® electroporation system. Nucleofection resulted in the production of IL-2 with a mean of 478.5 pg/106 cells (range of 107.6–1079.3 pg /106 cells/24 h) compared to mock transfected CIK cells (31 pg/106 cells) (P = 0.05). After co-culturing with DC their functional ability was assessed in vitro by a cytotoxicity assay. On comparison with non-transfected CIK cells co-cultured with DCs (36.5 ± 5.3 %), transfected CIK cells co-cultured with DC had a significantly higher lytic activity of 58.5 ± 3.2% (P = 0.03) against Dan G cells, a human pancreatic carcinoma cell line.