【摘要】 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。更多还原

【Abstract】 AIM:To explore the specific immunity of dendritic cell(DC)vaccine loading human umbilical vein endothelial cell(HUVEC)antigen.against U14 cervical cancer of mice.METHODS:Primary HUVECs were cultured,identified and made into antigen.The BALB/c mice were immuned with DCs loading HUVEC antigen 4 times a week.One week after last vaccination,U14 cervical cancer cells were injected subcutaneously into the mice.The tumor size,CTL response of spleen lymphocytes in vitro,the percentage of CD3+CD8+ surface markers of spleen lymphocytes,and the titer of serum antibody were detected.The specific immunity was examined by immunocytochemistry and Western blot.RESULTS:HUVECs with high purity were successfully cultured and by identified by immunocytochemistry and RT-PCR.After the vaccine was injected into mice,the tumors of mice in PBS group grew faster than the those in other groups.The tumors of mice in HUVEC-DC groups grew slowly and disappeared after 2 weeks and The tumors of mice in DC group disappeared after 3 weeks.The CTL response of spleen lymphocytes in vitro showed that HUVEC-DC-T cells could kill HUVEC cells.The percentage of CD3+CD8+ surface markers of spleen lymphocytes in HUVEC-DC group was higher than that in other groups.The titer of serum antibody was 1∶800.immunocytochemistry analysis indicated HUVEC-DC group had specific antigen-antibody reaction to HUVECs through and Western blot analysis revealed there were specific bands at 130 KDa and 220 KDa.CONCLUSION:HUVEC-DC vaccine and DC vaccine can inhibit the tumor growth of U14 cervical cancer of mice.The special cellular and humoral immunity are induced by HUVEC-DC vaccine.Furthermore,some antigens in HUVECs maybe have special immune reaction with integrin α ν and VEGF-R2.更多还原