Cancer gene therapy using in vivo electroporation of Flt3-ligand.
Cancer gene therapy using in vivo electroporation of Flt3-ligand.
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DOI：
10.3892/ijo.27.2.457
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发表时间：
2005-08
影响因子：
5.2
通讯作者：
Kazuya Shimao;T. Takayama;K. Enomoto;Tetsuya Saito;S. Nagai;J. Miyazaki;K. Ogawa;H. Tahara
中科院分区：
文献类型：
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作者：
Kazuya Shimao;T. Takayama;K. Enomoto;Tetsuya Saito;S. Nagai;J. Miyazaki;K. Ogawa;H. Tahara
树突状细胞（Dendritic cells，DC）是一种强有力的抗原提呈细胞，在免疫应答中起着关键作用。外周血单核细胞体外诱导的树突状细胞作为一种有前途的肿瘤免疫治疗手段已开始临床应用。如果不需要体外操作就可以获得相同类型的免疫刺激剂，那么在临床环境中可能会非常方便。在这项研究中，我们进行了系统的Flt 3L基因转移，在体内电穿孔Flt 3L质粒DNA（Flt 3L-IVE）在胫前肌，以确定对原位DC的影响。Flt 3L-IVE后，在IVE后10天的血清中检测到显著水平的Flt 3L。在Flt 3L-IVE后4天观察到血清中5326+/-920 pg/ml的峰浓度。与对照组相比，Flt 3L-IVE组脾脏和骨髓中DC数量显著增加，并显示出高水平的共刺激分子表达。免疫组化结果显示，与对照组相比，Flt 3L-IVE诱导后21 d，肿瘤组织中不仅有DC，而且有CD 8和CD 4阳性细胞浸润，并在肿瘤组织中存在。然而，Flt 3L-IVE的抗肿瘤作用在MCA 205建立的肿瘤中不显著。肿瘤浸润DC多为未成熟表型。外周区仅有少量DCs具有成熟表型。这些结果表明，使用体内电穿孔的Flt 3L基因转移可以动员DC进入肿瘤部位。诱导这些DC成熟的其他手段可能对该策略的抗肿瘤作用产生积极影响。
Dendritic cells (DCs) are potent antigen-presenting cells which play pivotal roles in immunological response. The clinical application of DCs induced from peripheral monocytes in vitro has been initiated as a promising immuno-logical therapy against cancer. If the same type of immuno-stimulator could be achieved without in vitro manipulation, it might be very convenient in clinical settings. In this study, we performed systemic gene transfer of Flt3L using in vivo electroporation of Flt3L plasmid DNA (Flt3L-IVE) in pretibial muscles in order to determine the effects on DCs in situ. After Flt3L-IVE, Flt3L was detected in the serum for 10 days after IVE at significant levels. The peak concentration of 5326+/-920 pg/ml in the serum was observed 4 days after Flt3L-IVE. The number of DCs was significantly increased and showed highly co-stimulatory molecule expressions both in spleen and bone marrow after Flt3L-IVE compared to those of control groups. Immunohistochemical evaluation revealed that not only DCs but also CD8 and CD4 positive cells were significantly infiltrated into the local tumor site compared with those of control and remained in the tumor 21 days after a single Flt3L-IVE. However, anti-tumor effects of Flt3L-IVE were not significant in the MCA205 established tumor. Most of the tumor infiltrating DCs had immature phenotype. Only a small number of DCs in the peripheral areas had the mature phenotype. These results suggest that Flt3L gene transfer using in vivo electroporation could mobilize DCs into tumor site. Additional means to induce maturation of these DCs could have a positive impact on anti-tumor effects of this strategy.