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目的构建重组EGFR噬菌体疫苗并且观察其抗肿瘤效果。方法将鸡源EGFR膜外部分的5个基因片段插入到T7噬菌体展示系统中,EGFR蛋白以融合蛋白的形式展示在噬菌体的外壳蛋白10B上,从而构建成噬菌体疫苗。Western blot方法检测噬菌体壳蛋白上是否有EGFR表达。利用ELISA方法检测免疫小鼠血清中抗EGFR抗体。分离免疫小鼠脾细胞,检测淋巴细胞对靶细胞A431的细胞毒作用。C57小鼠经过4次免疫,接种Lewis肺癌,记录肿瘤的生长情况,评价疫苗的抗肿瘤效果。结果本实验成功构建5种鸡源EGFR噬菌体疫苗。Western blot。显示噬菌体壳蛋白上有EGFR表达。免疫后的小鼠血清中检测到抗EGFR抗体。体外分离脾细胞证明免疫后的小鼠脾细胞对EGFR受体高表达的A431细胞具有杀伤作用。靶细胞和效应细胞的比例在1:10时,最大杀伤率可达(45．74±7．21)％(P<0．001)。肿瘤生长曲线显示免疫后的C57小鼠Lewis肺癌生长受到明显的抑制。结论利用噬菌体展示技术构建的噬菌体疫苗是一种有效的抗肿瘤方法。
Objective To construct recombinant EGFR phage vaccine and observe its anti-tumor effect. Methods Five gene fragments of the EGFR outer membrane were inserted into the T7 phage display system. The EGFR protein was expressed on the coat protein 10B of the phage as a fusion protein to construct a bacteriophage vaccine. Western blot method was used to detect the expression of phage coat protein EGFR. Anti-EGFR antibodies in serum of immunized mice were detected by ELISA. The splenocytes of immunized mice were isolated and the cytotoxic effect of lymphocytes on target cells A431 was tested. After four immunizations, C57 mice were inoculated with Lewis lung cancer, the growth of the tumor was recorded, and the anti-tumor effect of the vaccine was evaluated. Results Five chicken-derived EGFR phage vaccines were successfully constructed in this experiment. Western blot. EGFR expression on phage coat protein. Anti-EGFR antibodies were detected in the sera of immunized mice. Isolation of spleen cells in vitro demonstrated that the immunized mouse splenocytes have a killing effect on A431 cells with high EGFR receptor expression. The ratio of target cells to effector cells was 1:10, the maximum killing rate was (45.74 ± 7.21)% (P <0.001). Tumor growth curve showed that the growth of Lewis lung carcinoma in C57 mice was significantly inhibited after immunization. Conclusion The phage vaccine constructed by phage display technique is an effective anti-tumor method.