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目的:研究猪endoglin胞外段重组蛋白(pEDG)作为异种蛋白疫苗是否能够诱导小鼠产生抗自身endoglin的免疫反应,从而抑制endoglin相关的肿瘤血管生成,进而抑制肿瘤的生长。方法:观察免疫治疗的荷瘤小鼠肿瘤体积和生存情况,用免疫组化方法(抗CD31)观察肿瘤组织血管生长并计算肿瘤微血管密度,用免疫荧光方法了解肿瘤血管自身抗体沉积,用Westernblot和ELISA方法检测荷瘤小鼠免疫治疗后的血清是否含有抗自身endoglin的抗体和抗体亚型,用ELISPOT方法检测荷瘤小鼠脾脏中分泌抗endoglin抗体的B淋巴细胞。结果:与同种重组endoglin蛋白和非疫苗对照组二个对照组相比,重组pEDG蛋白疫苗主动免疫治疗可以明显抑制小鼠肿瘤的生长,小鼠生存时间明显延长,肿瘤组织中血管生成明显减少,肿瘤组织血管表面有自身抗体沉积,血清中含有抗自身endoglin的抗体,抗体亚型主要为IgG1和IgG2b。此外,免疫治疗组小鼠脾脏中还发现明显增多的分泌抗自身endoglin抗体的B淋巴细胞。结论:pEDG疫苗诱导抗自身endoglin抗体的产生,从而抑制肿瘤血管生成和肿瘤生长。这种主动免疫治疗的方法有望成为治疗肿瘤的新策略。
OBJECTIVE: To investigate whether the recombinant protein (pEDG) of pig endoglin as a heterologous protein vaccine can induce immune responses against endogenous endoglin in mice, thereby inhibiting endoglin-associated tumor angiogenesis and inhibiting tumor growth. METHODS: The tumor volume and survival of tumor-bearing mice treated with immunotherapy were observed. Tumor vascular growth was observed by immunohistochemistry (anti-CD31) and the tumor microvessel density was calculated. Tumor vascular autoantibody deposition was assessed by immunofluorescence using Western blot and ELISA method was used to test whether the serum of the tumor-bearing mice after the immunotherapy contained anti-self doglin antibody and antibody subtypes. The ELISPOT method was used to detect the B-lymphocytes secreting anti-endoglin antibody in the spleen of tumor-bearing mice. RESULTS: Compared with the same reconstructed endoglin protein and non-vaccine control groups, the active immunotherapy with the recombinant pEDG protein vaccine significantly inhibited the growth of mice. The survival time of mice was significantly prolonged, and the angiogenesis in the tumor tissue was significantly reduced. There is autoantibody deposition on the vascular surface of the tumor tissue. The serum contains antibodies against its own endoglin. The antibody subtypes are mainly IgG1 and IgG2b. In addition, significantly increased B lymphocytes secreting anti-self-endoglin antibodies were also found in the spleen of immunotherapy mice. CONCLUSION: The pEDG vaccine induces the production of antibodies against its own endoglin, thereby inhibiting tumor angiogenesis and tumor growth. This method of active immunotherapy is expected to become a new strategy for the treatment of tumors.