title: [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].
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+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
  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 lymphocytes in vitro showed that bEnd.3-DC cells could kill bEnd.3 cells, the special lysis rate was more than 60%. The percentage of CD₃+CD₈+ 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 bEnd.3-DC group was 1:3200, while it was 1:800 in DC group and there were not any in PBS group. 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 humoral 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.
authors.affiliation: Department of Pathophysiology, College of Basic Medical Sciences, Zhengzhou University, Zhengzhou 450001, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Zhao
authors.forename: Jing
authors.initials: J
authors.lastname: Lu
authors.forename: Ya-qin
authors.initials: YQ
authors.lastname: Liu
authors.forename: Hong-yan
authors.initials: HY
authors.lastname: Yang
authors.forename: You-tian
authors.initials: YT
authors.lastname: Huang
authors.forename: Ji-min
authors.initials: JM
authors.lastname: Zhao
authors.forename: Shan
authors.initials: S
authors.lastname: Li
authors.forename: Jing-ming
authors.initials: JM
authors.lastname: Zhai
authors.forename: Ming-yao
authors.initials: MY
authors.lastname: Zhao
authors.forename: Xi
authors.initials: X
authors.lastname: Zhang
authors.forename: Zi-ming
authors.initials: ZM
authors.lastname: Dong
mesh_terms: D000818:Animals; D000941:Antigens / Q000276:immunology*; D015703:Antigens, CD / Q000276:immunology; D019496:Cancer Vaccines / Q000276:immunology*; D045744:Cell Line, Tumor; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology* / Q000637:transplantation; D042783:Endothelial Cells / Q000276:immunology*; D005260:Female; D016219:Immunotherapy, Adoptive; D039564:Integrin alphaV / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013154:Spleen / Q000166:cytology / Q000276:immunology; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D002583:Uterine Cervical Neoplasms / Q000276:immunology* / Q000473:pathology / Q000628:therapy; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000276:immunology / Q000378:metabolism
chemical_list: D000941:Antigens; D015703:Antigens, CD; D019496:Cancer Vaccines; D039564:Integrin alphaV; D040301:Vascular Endothelial Growth Factor Receptor-2