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Clinical Application of Tumor-specific Anti-tumor Immunity Induced by Tumor Cells Expressing Fas (CD95) Ligand

表达Fas（CD95）配体的肿瘤细胞诱导肿瘤特异性抗肿瘤免疫的临床应用

基本信息

批准号：
13557097
负责人：
TAKEDA Yasutaka
金额：
$ 8.83万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

项目摘要

CD95 ligand (FasL)-expressing tumors cause immunopotentiation following vigorous neutrophil infiltration. Thus, the induction of neutrophil Infiltration by FasL appears to play an important role in tumor rejection. 1) The mechanism by which FasL-expressing tumors cause neutrophil infiltration has not been well understood. Here, we Investigated the role of chemokines in FasL-induced antitumor activity. CXC chemokine receptor 2 (CXCR2) knockout (KO) mice are a powerful tool for studying CXC chemokine-mediate neutrophil infiltration. Thus, we examined the mechanism for the neutrophil recruitment Induced by FasLcDNA-transfected MethA (MethA+FasL) fibrosarcoma using CXCR2 KO mice. MethA+FasL cells were completely rejected in wild-type (WT) and KO mice. MethA+FasL cells Injected i.p. induced the recruitment of both neutrophils and macrophages in peritoneal cavity (PC) of WT but only macrophages in PC of KO mice, although CXC and CC chemokines were released In PC in both mice. Macrophages Inc … More ubated with MethA+FasL cells released CXC and CC chemokines. F4/80-positive macrophages decreased after injection of MethA+FasL cells. Macrophages derived from WI and KO but not neutrophils from WT mice induced the recruitment of neutrophils when adoptively i.p. transferred with MethA+FasL cells into FasL/Fas-deficient mice. The different recruitment of inflammatory cells between WT and KO mice was attributed to bone marrow (BM) cells by BM transfer experiment. These results demonstrated that CXC chemokines are essential for neutrophil recruitment and that macrophages but not neutrophils play a critical role in FasL-induced infiltration of Inflammatory cells and that chemokines played important roles In eradication of FasL-expressing tumor cells and induction of antitumor immunity. 2) Mouse mammary tumor, BJMC3879, has highly metastatic potential. FasLcDNA-transfected BJMC3879 (BJMC+FasL) was established for clinical application of breast cancer. Although It induced neutrophil recruitment, It was not rejected In WT mice. The reason might be that BJMC+FasL was little expressed FasL-protein on cell surface. It could be indicated that the capability of FasL-protein expression depends on a kind of tumor cell. 3) On the other hand, we were developing non-viral vectors for clinical application. We are investigating the effects of Quarternary complex (Qplex) (cDNA+cationlc liposome+protamine+transferin) or polyethilenimine on transfecting rate of a few cell lines. Several problems for clinical application become clear. Less

表达CD 95配体（FasL）的肿瘤在剧烈的中性粒细胞浸润后引起免疫增强。因此，FasL诱导中性粒细胞浸润似乎在肿瘤排斥中起重要作用。1)表达FasL的肿瘤引起中性粒细胞浸润的机制尚未得到很好的理解。在这里，我们研究了趋化因子在FasL诱导的抗肿瘤活性中的作用。CXC趋化因子受体2（CXCR 2）敲除（KO）小鼠是研究CXC趋化因子介导的中性粒细胞浸润的有力工具。因此，我们使用CXCR 2 KO小鼠研究了由FasL cDNA转染的MethA（MethA+FasL）纤维肉瘤诱导的中性粒细胞募集的机制。MethA+FasL细胞在野生型（WT）和KO小鼠中被完全排斥。腹膜内注射MethA+FasL细胞诱导WT小鼠腹腔（PC）中中性粒细胞和巨噬细胞的募集，但KO小鼠PC中仅巨噬细胞的募集，尽管在两种小鼠的PC中均释放CXC和CC趋化因子。Macrophage Inc ...更多信息用MethA+FasL处理的细胞释放CXC和CC趋化因子。注射MethA+FasL细胞后，F4/80阳性巨噬细胞减少。当用MethA+FasL细胞过继腹膜内转移到FasL/Fas缺陷小鼠中时，来自WI和KO的巨噬细胞而不是来自WT小鼠的中性粒细胞诱导中性粒细胞的募集。通过骨髓转移实验，WT和KO小鼠之间炎性细胞的不同募集归因于骨髓（BM）细胞。这些结果表明，CXC趋化因子是必需的中性粒细胞募集和巨噬细胞，而不是中性粒细胞在FasL诱导的炎性细胞的浸润中发挥关键作用，趋化因子在根除表达FasL的肿瘤细胞和诱导抗肿瘤免疫中发挥重要作用。2)小鼠乳腺肿瘤BJMC 3879具有高度转移潜能。建立了转染FasL cDNA的BJMC 3879（BJMC+FasL）细胞系，为乳腺癌的临床应用奠定了基础。虽然它诱导中性粒细胞募集，但在WT小鼠中没有排斥。其原因可能是BJMC+FasL细胞表面很少表达FasL蛋白。说明FasL蛋白的表达能力取决于肿瘤细胞的种类。3)另一方面，我们正在开发用于临床应用的非病毒载体。我们研究了四元复合物（cDNA+阳离子脂质体+鱼精蛋白+转铁蛋白）或聚乙烯亚胺对几种细胞系转染率的影响。临床应用的几个问题变得清晰。少