【摘要】 目的评估OK432作为佐剂制备的内皮细胞疫苗体内抗肿瘤转移的作用。方法以OK432为佐剂,体外混合人脐静脉内皮细胞(HUVEC),制备HUVEC-OK432疫苗。雄性健康BALB/c小鼠尾静脉注射肝癌细胞5×105建立肝癌肺转移模型,皮下注射HUVEC-OK432疫苗,分别在预防性和治疗性免疫中评价HUVEC-OK432疫苗抗肿瘤转移活性;皮内注射5×104肝癌细胞建立皮内肿瘤血管模型,皮下注射HUVEC-OK432疫苗,评价HUVEC-OK432疫苗抑制肿瘤新生血管的活性;采用ELISA的方法测定小鼠免疫血清中HUVEC抗体水平;采用细胞毒T淋巴细胞(CTL)实验考察HUVEC-OK432疫苗诱导的细胞免疫应答水平。结果肺转移模型结果显示,在预防性和治疗性免疫中,HUVEC-OK432免疫均能有效降低小鼠肺上的肿瘤转移灶数目(P<0.01);皮内肿瘤血管模型的结果显示,HUVEC-OK432免疫能显著降低肿瘤新生血管的数目(116.5±20.6 vs 45.0±8.9,P<0.01);ELISA测定抗体的结果显示,HUVEC-OK432初次免疫后1周小鼠即产生了高滴度的特异性HUVEC抗体,随免疫次数的增加抗体水平不断升高,且该抗体在体外可以显著抑制HUVEC细胞的增殖;CTL实验的结果表明,HUVEC-OK432免疫组淋巴细胞体外有明显的特异性杀伤HUVEC的作用。结论 HUVEC-OK432疫苗免疫可以有效诱导小鼠产生靶向内皮细胞的免疫应答,从而有效抑制小鼠肝癌细胞转移。更多还原

【Abstract】 OBJECTIVE To assess the anti-metastasis effect of OK432 optimized viable human umbilical vein endothelial cells( HUVECs) vaccine in vivo. METHODS OK432,a penicillin-inactivated streptococcus preparation,was combined with HUVECs to prepare a HUVECs-OK432 vaccine. An experimental metastasis model by tail vein injection of 5 × 105H22 hepatocellular carcinoma cells was employed to evaluate the anti-metastatic efficiency of HUVECs-OK432 in both prophylactic and therapeutic procedures. An intradermal tumor model by intradermal injection of 5 × 104 tumor cells was established to evaluate the anti-angiogenesis efficiency of HUVECs-OK432. Humoral immune responses were investigated using ELISA. Cellular immune responses were detected using a nonradioactive cytotoxicity assay.RESULTS In the in vivo lung metastasis model,HUVECs-OK432 immunization significantly reduced the number of lung nodules compared with PBS in both prophylactic and therapeutic procedures( P < 0. 01).Meanwhile,in the intradermal tumor model,HUVECs-OK432 could also decrease tumor-induced angiogenesis( 116. 5 ± 20. 6 vs 45. 0 ± 8. 9,P < 0. 01). HUVEC-specific antibodies,which further displayed a strong inhibition of HUVEC proliferation in vitro,were elevated in murine sera one week after the first immunization. In vitro analysis of splenocytes separeted from immunized mice revealed an induction of cytotoxic T lymphocytes with a lytic activity against activated endothelium. CONCLUSION HUVECsOK432 immunization can elicit both humoral and cellular immunity against active endothelium rather than tumor cells,and therefore significantly inhibit pulmonary metastasis of H22 hepatocellular carcinoma.更多还原