title: Enhanced induction of anti-tumor CTLs in vitro by a lentivirus-transduced dendritic cell vaccine expressing secondary lymphoid tissue chemokine and mucin 1.
abstract: OBJECTIVE
  Dendritic cell (DC)-based cancer immunotherapy requires an immunogenic tumor associated antigen (TAA) and an effective strategy for its presentation to lymphocytes. Here, we explored whether transduction of DCs with lentiviruses (LVs) expressing a fusion protein of secondary lymphoid tissue chemokine (SLC) and mucin 1 (MUC1) could stimulate antigen-specific cytotoxic T cells (CTLs) to human cancer cells in vitro.
  
  
  METHODS
  HLA-A2+ peripheral blood monocyte-derived DCs were transduced with recombinant lentiviruses LV at different multiplicities of infection (MOI), and MUC1, SLC or SLC-MUC1 mRNA and protein were detected by RT-PCR and Western blotting, respectively. Transduction efficiencies and phenotypes of DCs were evaluated by flow cytometry. Induction of T lymphocyte proliferation by DCs was examined with a Cell Count Kit-8 (CCK-8). CTL activities against tumor cells were analyzed by lactate dehydrogenase (LDH) cytotoxicity and enzyme-linked immunospot (ELISPOT) assays.
  
  
  RESULTS
  Stable expression of MUC1, SLC and SLC-MUC1 was obtained in DCs transduced with recombinant LVs, and the transduction efficiencies were dose-dependent. Transduction with LVs did not appreciably change the DC phenotype. CTL induced by LV MUC1 DCs potently and specifically lysed the HLA-A2+, MUC1+colon cancer cell line HCT-116. Moreover, this cytolytic activity against HCT-116 was enhanced with CTL stimulated by LV SLC-MUC1 DCs.
  
  
  CONCLUSIONS
  DCs transduced with MUC1 could induce effective cytolytic activity against tumor cells in an antigen-specific and HLA-restricted fashion in vitro, and SLC promoted MUC1-specific anti-tumor activity. The transduction of DCs with LV SLC- MUC1 may be a promising strategy in DC-based cancer immunotherapy.
authors.affiliation: Department of General Surgery, First Affiliated Hospital, Nanjing Medical University, Nanjing, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Wang
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhu
authors.forename: Jing-jing
authors.initials: JJ
authors.lastname: Zhang
authors.forename: Zheng
authors.initials: Z
authors.lastname: Chen
authors.forename: Wen-biao
authors.initials: WB
authors.lastname: Liang
authors.forename: Ze-kuan
authors.initials: ZK
authors.lastname: Xu
mesh_terms: D019496:Cancer Vaccines / Q000235:genetics / Q000276:immunology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D054421:Chemokine CCL21 / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D003110:Colonic Neoplasms / Q000276:immunology / Q000628:therapy*; D003713:Dendritic Cells / Q000276:immunology* / Q000378:metabolism; D018014:Gene Transfer Techniques; D006801:Humans; D007167:Immunotherapy; D016086:Lentivirus / Q000235:genetics; D008213:Lymphocyte Activation; D007959:Lymphocyte Culture Test, Mixed / Q000379:methods; D018396:Mucin-1 / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014162:Transfection
chemical_list: C515478:CCL21 protein, human; D019496:Cancer Vaccines; D054421:Chemokine CCL21; C060192:MUC1 protein, human; D018396:Mucin-1