Study on the specific immunity induced by dendritic cell vaccine loading allogenic microvascular endothelial cell bEnd. 3 antigen against U14 cervical cancer cell in mice / 中华妇产科杂志

ABSTRACT

Objective To explore the specific cellular and humoral immunity induced by dendritic cells (DC) vaccine loading allogenic microvascular endothelial cell bEnd. 3 antigen against U14 cervical cancer cell of mice. Methods Mouse brain microvascular endothelial cell bEnd. 3 was cultured and identified for preparation endothelial cell bEnd. 3 antigen. The level of mRNA expression of vascular endothelial growth factor receptor 2 (VEGF-R2) and integrin αV was detected by reverse transcription (RT)-PCR. The BALB/c mice were immuned with DC loading bEnd. 3 antigen 4 times in 4 weeks (bEnd. 3-DC group), while the mice only were immuned with DC or injected with phosphate buffer saline (PBS group) as control group. One week after last vaccination, U14 cervical cancer cells were injected subcutaneously into the mice. The tumor size, cytotoxic T lymphocyte (CTL) response of spleen lymphocytes in vitro, the percentage of CD3+ CD+8 surface markers of spleen lymphocytes, and the titer of serum antibody were detected. The specific immunity was examined by immunocytochemistry and western blot. Results The expression of VEGF-R2 and integrin αV gene in bEnd. 3 cells were expressed highly.After the vaccine was injected, the tumors of mice in PBS group grew faster than those in other groups, while the tumors in bEnd. 3-DC group grew slowly and disappeared after 2 weeks. The volume of tumors in DC group grew slower than those in PBS group [(0.11± 0.13) cm3 versus (3.38 ±0.34) cm3]. The CTL response of spleen iymphocytes in vitro showed that bEnd. 3-DC cells could kill bEnd. 3 cells, the special lysis rate was more than 60% . The percentage of CD+3 CD+8 spleen lymphocytes in bEnd. 3-DC group[(38.6 ± 0.7) %] was higher than those in other groups (P ＜ 0.05). The titer of serum antibody of Immunocytochemistry analysis indicated there were specific antigen-antibody reaction to bEnd. 3 cell in bEnd. 3-DC group. Western blot analysis revealed that there were specific bands at 220 000 (VEGF-R2).Conclusions bEnd. 3-DC vaccine can inhibit the tumor growth of U14 cervical cancer cell of mice, which indicates that the special cellular and humorai immunity are induced by bEnd. 3-DC antigen which maybe have some antigens in bEnd. 3 cells that reacts with endothelial cell proliferation-related antigens.