Induction of IFN-regulated factors and antiturnoral surveillance by transfected placebo plasmid DNA
Induction of IFN-regulated factors and antiturnoral surveillance by transfected placebo plasmid DNA
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DOI：
10.1016/j.ymthe.2004.09.008
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发表时间：
2005-01-01
影响因子：
12.4
通讯作者：
Bhardwaj, A
中科院分区：
文献类型：
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作者：
Li, SL;Wilkinson, M;Bhardwaj, A
体内递送编码治疗基因的DNA对于治疗恶性肿瘤以及遗传疾病具有巨大的潜力。在基因治疗研究中，使用不含转基因的安慰剂DNA的递送作为对照。大多数研究者默认，从转染的DNA表达的蛋白质具有表型后果，但这些后果不是来自DNA本身。在这里，我们证明了将对照质粒DNA（不表达基因产物）转染到肿瘤细胞系中诱导肿瘤细胞系中干扰素（IFN）调节基因IRF 7、STAT 1、CD 11（批准的基因符号CXCL 9）、MHCI（云母）和CD 11 a（ITGAL）的表达显著增加（> 10倍）。这些基因的诱导抑制用凋亡肿瘤细胞免疫的免疫活性小鼠中的肿瘤发展和肿瘤生长。抗体耗尽研究表明，对照DNA的转染诱导IFN调节基因的潜在机制是诱导分泌因子，例如IFN-β。三条证据表明，DNA转染介导的IFN-调节基因的诱导是独立的TLR 9。这三条证据是：（1）TLR 9在SCCVII或4 T1细胞系中均不表达，（2）TLR 9下游信号分子的活化与基因表达的诱导无关，和（3）从DNA转染的SCCVII肿瘤细胞的条件培养基中获得的分泌因子在4 T1肿瘤细胞系中诱导相同类型的基因表达，其对DNA转染的基因诱导是抗性的。我们的发现表明，4 T1肿瘤细胞系，这是抵抗DNA转染介导的诱导IFN-调节基因，可以用来确定真实的治疗基因的功能。
Delivery of DNA encoding therapeutic genes in vivo has great potential for treating malignancy as well as genetic diseases. Delivery of placebo DNA without a transgene is used as a control in gene therapy studies. It is tacitly assumed by most investigators that the protein expressed from the transfected DNA has phenotypic consequences, but that the consequences are not from the DNA itself. Here, we demonstrate that transfection of control plasmid DNA (that does not express a gene product) into tumor cell lines induces a dramatic (> 10-fold) increase in the expression of the interferon (IFN)-regulated genes IRF7, STAT1, MIG (approved gene symbol CXCL9), MHCI (MICA), and CD11a (ITGAL) in tumor cell lines. Induction of these genes inhibits tumor development and tumor growth in immunocompetent mice that are immunized with apoptotic tumor cells. The antibody depletion study indicates that the underlying mechanism by which transfection of control DNA induces IFN-regulated genes is the induction of a secreting factor(s) such as IFN-beta. Three lines of evidence indicate that DNA transfection-mediated induction of IFN-regulatory genes is independent of TLR9. The three lines of evidence are: (1) TLR9 is not expressed in either SCCVII or 4T1 cell line, (2) activation of TLR9 downstream signaling molecules is not associated with the induction of gene expression, and (3) the secretion factor(s) obtained from the conditioned medium of DNA-transfected SCCVII tumor cells induces the same type of gene expression in the 4T1 tumor cell line, which is refractory to the gene induction by DNA transfection. Our finding indicates that the 4T1 tumor cell line, which is resistant to the DNA transfection-mediated induction of IFN-regulated genes, can be used to determine the real therapeutic gene function.