Nagaraj Srinivas, Ziske Carsten, Schmidt-Wolf Ingo Gh
Department of Internal Medicine I, General Internal Medicine Rheinische Friedrich-Wilhelms-Universität, Bonn, Germany.
Genet Vaccines Ther. 2004 Aug 25;2:12. doi: 10.1186/1479-0556-2-12. eCollection 2004.
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 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/10 cells (range of 107.6-1079.3 pg /10 cells/24 h) compared to mock transfected CIK cells (31 pg/10 cells) ( = 0.05). After co-culturing with DC their functional ability was assessed 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% ( = 0.03) against Dan G cells, a human pancreatic carcinoma cell line.
通过基因改造的免疫效应细胞调节免疫系统在恶性肿瘤治疗中具有潜在的治疗价值。白细胞介素-2（IL-2）是一种关键的细胞因子，可诱导有效的抗肿瘤反应。细胞因子诱导的杀伤细胞（CIK）已被描述为高效的细胞毒性效应细胞，能够裂解肿瘤细胞靶标，并且能够以非主要组织相容性复合体（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。本研究评估了用IL-2进行非病毒转染并与脉冲和未脉冲的DC共培养的CIK细胞的抗肿瘤作用。从外周血中产生的人CIK细胞用编码人IL-2的质粒转染。转染涉及电参数和特定溶液的组合，以通过使用Nucleofector电穿孔系统将质粒直接递送至细胞核。与mock转染的CIK细胞（31 pg/10个细胞）相比，核转染导致IL-2的产生，平均为478.5 pg/10个细胞（范围为107.6 - 1079.3 pg /10个细胞/24小时）（P = 0.05）。与DC共培养后，通过细胞毒性试验评估它们的功能能力。与与DC共培养的未转染CIK细胞（36.5±5.3％）相比，与DC共培养的转染CIK细胞对人胰腺癌细胞系Dan G细胞具有明显更高的裂解活性，为58.5±3.2％（P = 0.03）。
