Zhao Jun, Zhai Jing-ming, Lu Jing, Yang Hong-yan, Huang You-tian, Li Shan, Dong Zi-ming, Zhang Xi
Department of Pathophysiology, College of Basic Medical Sciences, Zhengzhou University, Zhengzhou, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2009 Mar;25(3):196-200.
To explore the specific immunity of dendritic cell (DC) vaccine loading human umbilical vein endothelial cell (HUVEC) antigen.against U14 cervical cancer of mice.
Primary HUVECs were cultured, identified and made into antigen. The BALB/c mice were immunized 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.
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.
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 alphav and VEGF-R2.
探讨负载人脐静脉内皮细胞（HUVEC）抗原的树突状细胞（DC）疫苗对小鼠U14宫颈癌的特异性免疫作用。
培养、鉴定原代HUVECs并制成抗原。将负载HUVEC抗原的DCs每周免疫BALB/c小鼠4次。末次免疫1周后，将U14宫颈癌细胞皮下注射到小鼠体内。检测肿瘤大小、脾淋巴细胞体外CTL反应、脾淋巴细胞CD3(+)CD8(+)表面标志物百分比及血清抗体效价。通过免疫细胞化学和蛋白质印迹法检测特异性免疫。
成功培养出高纯度的HUVECs，并通过免疫细胞化学和RT-PCR进行鉴定。疫苗注射到小鼠体内后，PBS组小鼠肿瘤生长比其他组快。HUVEC-DC组小鼠肿瘤生长缓慢，2周后消失，DC组小鼠肿瘤3周后消失。脾淋巴细胞体外CTL反应显示HUVEC-DC-T细胞可杀伤HUVEC细胞。HUVEC-DC组脾淋巴细胞CD3(+)CD8(+)表面标志物百分比高于其他组。血清抗体效价为1:800。免疫细胞化学分析表明HUVEC-DC组对HUVECs有特异性抗原抗体反应，蛋白质印迹分析显示在130 KDa和220 KDa处有特异性条带。
HUVEC-DC疫苗和DC疫苗可抑制小鼠U14宫颈癌的肿瘤生长。HUVEC-DC疫苗可诱导特殊的细胞免疫和体液免疫。此外，HUVECs中的一些抗原可能与整合素αv和VEGF-R2有特殊免疫反应。
