title: [Changes in immune function of dendritic cells infected by recombinant adenovirus containing Her2/neu gene of extracellular and transmembrane domain proteins].
abstract: OBJECTIVE
  To observe the functional changes of dendritic cells (DCs) infected in vitro by 3 recombinant adenoviruses encoding Her2/neu extracellular first-receptor domain (Her2-ECDs), full-length extracellular domain (Her2-ECD), and extracellular and transmembrane domain (Her2-TM) proteins (rAdHer2-ECDs, rAdHer2-ECD and rAdHer2-TM, respectively).
  
  
  METHODS
  The expressions of the target proteins were detected with Western blotting. The level of both interleukin (IL)-12 in the supernatant of in vitro cultured DCs infected with recombined adenoviruses and interferon gamma (IFN-gamma) in the supernatant of the lymphocyte populations co-cultured with DCs were determined by enzyme-linked immunosorbent assay (ELISA). The capacity of the DCs to stimulate allogeneic T lymphocyte proliferation was assessed by mixed lymphocyte reaction, and the activity of cellular toxic T lymphocytes (CTL) were investigated by MTT assay.
  
  
  RESULTS
  Her2-ECDs, ECD and TM proteins were detected in the transfected DCs. Compared with the untransfected DCs, more abundant IL-12 production was detected in the supernatant of the DCs 5 days after transfection, but the IL-12 level showed no significant difference between the DCs infected with the 3 recombinant adenoviruses. IFN-gamma production increased gradually with passage of the time following DC-stimulated lymphocyte proliferation irrespective of infection of the DCs, and only the DCs infected with rAdHer2-TM seemed to result in significant difference in DC-mediated allogeneic T lymphocyte proliferation. The killing of breast cancer cell line with Her2 overexpression was more efficient with infected DCs priming autologous T lymphocyte to generate CTL than with uninfected DCs and those modified by SK-OV-3 cell fragment. CTL activity induced by rAdHer2-TM-infected DCs was the strongest, and breast cancer cell-killing activity was more efficient against cell line with Her2/neu-overexpression.
  
  
  CONCLUSIONS
  The DCs infected with the recombinant adenovirus encoding Her2/neu extracellular and transmembrane domains show enhanced anti-tumor effect and induce Her2/neu-specific CTL activity.
authors.affiliation: Department of Oncology, Institute of Tropical Medicine, Southern Medical University, Guangzhou 501515, China. mashudong@yahoo.com.cn
authors.forename: Shu-dong
authors.initials: SD
authors.lastname: Ma
authors.forename: Rong-cheng
authors.initials: RC
authors.lastname: Luo
authors.forename: Zhen-hua
authors.initials: ZH
authors.lastname: Ding
authors.forename: Feng
authors.initials: F
authors.lastname: Lu
authors.forename: Chang-qing
authors.initials: CQ
authors.lastname: Yuan
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000235:genetics / Q000276:immunology / Q000473:pathology; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002470:Cell Survival / Q000276:immunology; D018920:Coculture Techniques; D003602:Cytotoxicity, Immunologic / Q000276:immunology; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D016326:Extracellular Matrix Proteins / Q000235:genetics* / Q000378:metabolism; D005260:Female; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D018664:Interleukin-12 / Q000378:metabolism; D007959:Lymphocyte Culture Test, Mixed; D008214:Lymphocytes / Q000166:cytology / Q000276:immunology; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D010051:Ovarian Neoplasms / Q000235:genetics / Q000276:immunology / Q000473:pathology; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000166:cytology / Q000276:immunology; D014162:Transfection
chemical_list: D016326:Extracellular Matrix Proteins; D007371:Interferon-gamma; D018664:Interleukin-12; D008565:Membrane Proteins; D018719:Erb-b2 Receptor Tyrosine Kinases; D011993:Recombinant Fusion Proteins