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目的:探讨负载人脐静脉内皮细胞(HUVEC)抗原的DC疫苗诱导小鼠对宫颈癌U14的特异性免疫反应。方法:原代培养和鉴定人HUVECs,制备抗原,负载树突状细胞(DC),免疫小鼠,免疫结束后1周小鼠皮下注射小鼠宫颈癌U14细胞,观察肿瘤生长大小、检测免疫后小鼠脾细胞体外CTL效应、表面CD3+CD8+百分率和血清抗体滴度,通过细胞免疫组化和Western blot血清特异性反应分析。结果:经免疫组化鉴定成功培养出纯度高的HUVECs,抗原负载DC并注射BALB/c小鼠后,PBS组肿瘤生长最快,DC组和HUVEC-DC组肿瘤也有生长,但生长较慢,在2周后HUVEC-DC组肿瘤消失,3周后DC组肿瘤消失。小鼠脾细胞体外CTL结果显示HUVEC-DC组有明显的特异性杀伤HUVEC的作用。CD3+CD8+百分比提示HUVEC-DC组细胞毒性T细胞比例较其他两组高。小鼠血清抗体滴度示有抗体产生,免疫组化检测示组与HUVEC有特异性抗原抗体反应,Western blot显示在相对分子质量(Mr)130000和220000处有特异性条带。结论:负载人HUVECs抗原的DC疫苗和DC疫苗对小鼠宫颈癌U14有抑制作用,HUVEC-DC疫苗诱导了小鼠的细胞和体液免疫,并诱导了针对HUVECs细胞中的某些抗原的应答,这些抗原可能是整合素αν和VEGF-R2。
Objective: To investigate the specific immune response of cervical cancer U14 induced by DC vaccine loaded with human umbilical vein endothelial cells (HUVEC) antigen. Methods: Primary cultured and identified human HUVECs were used to prepare antigens and dendritic cells (DCs). Immunized mice were injected subcutaneously with mouse U14 cells one week after the immunization to observe the growth of tumor cells. In vitro CTL effect, percentage of surface CD3 + CD8 + and antibody titer of murine splenocytes were analyzed by immunohistochemistry and Western blot. Results: HUVECs with high purity were successfully cultured and identified by immunohistochemistry. After DCs were injected into BALB / c mice and injected with BALB / c mice, the tumor grew fastest in PBS group and the tumors in DC group and HUVEC-DC group grew slowly but grew slowly. After 2 weeks, the tumor of HUVEC-DC group disappeared, and the tumor of DC group disappeared after 3 weeks. In vitro CTL results of mouse spleen cells showed that the HUVEC-DC group had obvious specific anti-HUVEC effect. CD3 + CD8 + percentage prompted HUVEC-DC group of cytotoxic T cells than the other two groups. Antibody production was observed in mouse serum antibody titers. Immunohistochemistry showed that the group had specific antigen-antibody reaction with HUVEC. Western blot showed specific bands at molecular weight (Mr) of 130,000 and 220,000. CONCLUSION: DC vaccines and DC vaccines that are loaded with human HUVECs antigens have inhibitory effects on U14 in mice. HUVEC-DC vaccine induces cellular and humoral immunity in mice and induces responses against certain antigens in HUVECs. These antigens may be integrins αν and VEGF-R2.