T4 噬菌体表面展示 mVEGFR2 疫苗抑制肺癌血管生成。

Inhibition of tumor angiogenesis in lung cancer by T4 phage surface displaying mVEGFR2 vaccine.

机构信息

Department of Clinical Pharmacy, China Pharmaceutical University, Nanjing 210009, China.

出版信息

Vaccine. 2011 Aug 5;29(34):5802-11. doi: 10.1016/j.vaccine.2011.03.051. Epub 2011 Apr 9.

DOI:10.1016/j.vaccine.2011.03.051

Abstract

Vascular endothelial growth factor (VEGF) has been known as a potential vasculogenic and angiogenic factor and its receptor (VEGFR2) is a major receptor to response to the angiogenic activity of VEGF. The technique that to break the immune tolerance of "self-antigens" associated with angiogenesis is an attractive approach for cancer therapy with T4 phage display system. In this experiment, mouse VEGFR2 was constructed on T4 phage nanometer-particle surface as a recombinant vaccine. T4-mVEGFR2 recombinant vaccine was identified by PCR and western blot assay. Immunotherapy with T4-mVEGFR2 was confirmed by protective immunity against Lewis lung carcinoma (LLC) in mice. The antibody against mVEGFR2 was detected by ELISPOT, ELISA and Dot ELISA. The inhibitive effects against angiogenesis were studied using CD31 and CD105 via histological analysis. VEGF-mediated endothelial cells proliferation and tube formation were inhibited in vitro by immunoglobulin induced by T4-mVEGFR2. The antitumor activity was substantiated from the adoptive transfer of the purified immunoglobulin. Antitumor activity and autoantibody production of mVEGFR2 could be neutralized by the depletion of CD4+T lymphocytes. These studies strongly suggest that T4-mVEGFR2 recombinant vaccine might be a promising antitumor approach.

摘要

血管内皮生长因子（VEGF）已被认为是一种潜在的血管生成和血管生成因子，其受体（VEGFR2）是对 VEGF 的血管生成活性产生反应的主要受体。打破与血管生成相关的“自身抗原”免疫耐受的技术是癌症治疗的一种有吸引力的方法，采用 T4 噬菌体展示系统。在该实验中，构建了小鼠 VEGFR2 作为重组疫苗在 T4 噬菌体纳米颗粒表面。通过 PCR 和 Western blot 鉴定 T4-mVEGFR2 重组疫苗。通过 T4-mVEGFR2 免疫疗法在小鼠中对 Lewis 肺癌（LLC）的保护性免疫进行了确认。通过 ELISPOT、ELISA 和 Dot ELISA 检测针对 mVEGFR2 的抗体。通过组织学分析，通过 CD31 和 CD105 研究了对血管生成的抑制作用。体外实验中，T4-mVEGFR2 诱导的免疫球蛋白抑制了 VEGF 介导的内皮细胞增殖和管腔形成。通过纯化免疫球蛋白的过继转移证实了抗肿瘤活性。mVEGFR2 的抗肿瘤活性和自身抗体产生可通过耗尽 CD4+T 淋巴细胞来中和。这些研究强烈表明 T4-mVEGFR2 重组疫苗可能是一种很有前途的抗肿瘤方法。