title: An autologous therapeutic dendritic cell vaccine transfected with total lung carcinoma RNA stimulates cytotoxic T lymphocyte responses against non-small cell lung cancer.
abstract: OBJECTIVE
  The development of immunotherapy for malignancy is greatly limited by the characteristic weak antigenicity of tumors. The primary goal of this study was to circumvent the isolation and purification of tumor-specific antigen determinants by producing a vaccine using lung tumor RNA-loaded dendritic cells (DCs), and to test the response against lung cancer.
  
  
  METHODS
  Total RNA was isolated from 18 lung carcinomas with positive carcinoembryonic antigen (CEA) and mucin-1 (MUC1) staining, as identified by immunohistochemistry. DCs and T-cells from peripheral blood mononuclear cells were generated in vitro, and then DCs in different stages were transfected with RNA using several different methods. The expression of CEA and MUC1 in RNA-transfected DCs was measured using flow cytometry. T-cells stimulated by DCs were harvested as effectors, and primary tumor cells cultured in vitro were used as targets. Cytotoxicity was determined by lactic dehydrogenase detection assay.
  
  
  RESULTS
  Immature RNA-transfected DCs significantly increased the expression of CEA and MUC1, compared to mature transfected DCs. RNA transfection via electroporation resulted in significantly greater CEA and MUC1 expression than did transfection via lipofection or passive pulsing. Lymphocytes stimulated by DCs transfected with lung tumor RNA initiated a cytotoxic T lymphocyte (CTL) tumor-specific response.
  
  
  CONCLUSIONS
  Immature DCs transfected with total lung carcinoma RNA by electroporation in vitro effectively stimulate antigen-specific CTL responses against tumor cells.
authors.affiliation: Guangdong Lung Cancer Institute, Guangdong General Hospital, Guangdong Academic of Medical Sciences, Guangzhou, 510080, PR China. gzwangkun@126.com
authors.forename: Kun
authors.initials: K
authors.lastname: Wang
authors.forename: Qing
authors.initials: Q
authors.lastname: Zhou
authors.forename: Ai-Lin
authors.initials: AL
authors.lastname: Guo
authors.forename: Chong-Rui
authors.initials: CR
authors.lastname: Xu
authors.forename: She-Juan
authors.initials: SJ
authors.lastname: An
authors.forename: Yi-Long
authors.initials: YL
authors.lastname: Wu
mesh_terms: D000328:Adult; D000368:Aged; D019496:Cancer Vaccines / Q000276:immunology*; D002272:Carcinoembryonic Antigen / Q000032:analysis / Q000235:genetics; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D002470:Cell Survival; D003602:Cytotoxicity, Immunologic / Q000276:immunology; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D018274:Electroporation; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D016130:Immunophenotyping; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D008175:Lung Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D008297:Male; D008875:Middle Aged; D018396:Mucin-1 / Q000032:analysis / Q000235:genetics; D012334:RNA, Neoplasm / Q000235:genetics*; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014162:Transfection / Q000379:methods; D014407:Tumor Cells, Cultured
chemical_list: D019496:Cancer Vaccines; D002272:Carcinoembryonic Antigen; C060192:MUC1 protein, human; D018396:Mucin-1; D012334:RNA, Neoplasm