Suppression of choroidal neovascularization by dendritic cell vaccination targeting VEGFR2.
Suppression of choroidal neovascularization by dendritic cell vaccination targeting VEGFR2.
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DOI：
10.1167/iovs.07-0425
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发表时间：
2007-10
影响因子：
4.4
通讯作者：
H. Mochimaru;N. Nagai;Go Hasegawa;C. Kudo-Saito;T. Yaguchi;Y. Usui;T. Kurihara;Takashi Koto;S. Satofuka;H. Shinoda;Y. Ozawa;K. Tsubota;Y. Kawakami;S. Ishida
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文献类型：
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作者：
H. Mochimaru;N. Nagai;Go Hasegawa;C. Kudo-Saito;T. Yaguchi;Y. Usui;T. Kurihara;Takashi Koto;S. Satofuka;H. Shinoda;Y. Ozawa;K. Tsubota;Y. Kawakami;S. Ishida
目的探讨诱导血管内皮生长因子受体(VEGFR)2细胞免疫是否抑制脉络膜新生血管(CNV)的形成。方法以VEGFR2的400~408位氨基酸对应的H-2DB限制性多肽作为表位多肽。从C57BL/6小鼠骨髓祖细胞中分离出树突状细胞(DC)。6周龄C57BL/6小鼠每隔6天皮下注射表位多肽致敏的成熟树突状细胞三次。第三次免疫后，行激光光凝诱导新生血管形成。光凝后1周处死小鼠，采集脉络膜和脾细胞。通过体积测量评估CNV体积。为确定表位多肽在C57BL/6小鼠体内的特异性免疫原性，从采集的脾细胞中分离CD8T细胞，用酶联免疫斑点法和酶联免疫吸附试验检测干扰素-γ和肿瘤坏死因子-α的产生。为了确定负责免疫治疗的T细胞亚群，小鼠被腹腔注射抗CD4或抗CD8耗竭抗体。结果经VEGFR2表位多肽免疫的小鼠新生血管体积明显小于未免疫或对照多肽gp70免疫的小鼠。细胞因子分析显示CD8T细胞以剂量依赖方式特异性产生干扰素-γ和肿瘤坏死因子-α。在体内，CD8而不是CD4，T细胞的耗尽显著逆转了VEGFR2多肽冲击的DC疫苗对CNV的抑制作用，达到了在未免疫或gp70免疫的动物中观察到的水平。结论VEGFR2多肽特异性诱导的细胞免疫通过CD8 T细胞的细胞毒作用抑制CNV。本研究结果提示以VEGFR2为靶点的DC疫苗作为治疗CNV的一种新策略的可能性。
PURPOSE To investigate whether the induction of cellular immunity against vascular endothelial growth factor receptor (VEGFR) 2 inhibits the development of choroidal neovascularization (CNV). METHODS H-2Db-restricted peptide corresponding to amino acids 400 to 408 of VEGFR2 was used as an epitope peptide. Dendritic cells (DCs) were harvested from bone marrow progenitors of C57BL/6 mice. Six-week-old C57BL/6 mice received subcutaneous injections of the epitope peptide-pulsed mature DCs three times at 6-day intervals. After the third immunization, laser photocoagulation was performed to induce CNV. One week after photocoagulation, mice were killed to harvest the choroid and splenocytes. CNV volume was evaluated by volumetric measurements. To confirm the specific immunogenicity of the epitope peptides in C57BL/6 mice, CD8 T cells isolated from harvested splenocytes were restimulated to measure interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha production through enzyme-linked immunospot assay and ELISA. To determine the T-cell subset responsible for the immunotherapy, mice were intraperitoneally injected with an anti-CD4 or anti-CD8 depletion antibody. RESULTS CNV volume was significantly lower in mice immunized with the VEGFR2 epitope peptide than in those not immunized or immunized with a control peptide gp70. Cytokine assays showed the peptide-specific production of IFN-gamma and TNF-alpha from the CD8 T cells in a dose-dependent manner. In vivo depletion of CD8, but not CD4, T cells significantly reversed the suppressive effect of the VEGFR2 peptide-pulsed DC vaccination on CNV to the level observed in nonimmunized or gp70-immunized animals. CONCLUSIONS These results indicate that the VEGFR2 peptide-specific induction of cellular immunity inhibits CNV through the cytotoxicity of CD8 T cells. Results of the present study suggested the possibility of DC vaccination targeting VEGFR2 as a novel therapeutic strategy for CNV.