title: Carboxymethylpachymaran enhances immunologic function of dendritic cells cultured in two kinds of hepatoma carcinoma cell line's supernatant via nuclear factor κ B/Rel pathway.
abstract: OBJECTIVE
  To study the immunologic function of dendritic cells (DCs) cultured in two kinds of hepatoma cell line's supernatant and the enhancing effects of carboxymethylpachymaran (CMP) on DCs.
  
  
  METHODS
  DCs were harvested after stimulation by granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-4 from umbilical cord blood using density-gradient centrifugation method. Cultured supernatant of two hepatoma cell lines (HepG2 and HepG2.2.15) were collected for condition medium (CM) according to a volume ratio of supernatant to incomplete RPMI-1640 medium, which was 3:1. CMP was dissolved in incomplete RPMI-1640 medium. Experimental groups were divided according to the culture medium, either CM or with CMP in it. DCs subsets CD83, CD86, CD1a, and d-related human leukocyte antigens (HLA-DR) were analyzed by flow cytometry. The proliferation ability of allogeneic T cells in mixed lymphocyte reaction (MLR) stimulated by DCs was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) analysis. IL-12p70, interferon-γ (IFN-γ), and nuclear factor κB (NF-κB) were detected by enzyme-linked immunosorbent assay analysis.
  
  
  RESULTS
  The proliferation of lymphocytes and secreting level of IL-12 and expression of phenotype of DCs cultured in two kinds of CM were lower than those of normal group (P <0.01). Compared with the normal group, groups treated with CMP showed a higher expression level of DCs subsets, lymphocyte reproductive activity, as well as IL-12 and IFN-γ secretion levels. Groups treated with CMP also demonstrated higher levels of DCs phenotype expression and IL-12 and IFN-γ secretion in supernatant of MLR and higher lymphocyte reproductive activity compared with CM group (P <0.05). Compared with the normal group, the expression level of NF-κB in DCs nuclear was higher in CMP groups but lower in two CM groups (P <0.05). After CMP was added, the NF-κB expression levels of two CM groups were increased compared with levels before CMP was added (P <0.05). However, there was no significant difference between the two CM groups (P >0.05).
  
  
  CONCLUSIONS
  Two kinds of hepatoma cell line's supernatant can inhibit the immunologic function of DCs. This suppressive effect may be related to the inhibition of NF-κB/Rel pathway. CMP may up-regulate the DCs function by activating the NF-κB/Rel pathway.
authors.affiliation: Medical College, Jinan University, Guangzhou 510632, China.
authors.forename: Zhuo
authors.initials: Z
authors.lastname: Chen
authors.forename: Bin
authors.initials: B
authors.lastname: Yu
authors.forename: Xian-Lin
authors.initials: XL
authors.lastname: Wu
authors.forename: Cong-Qi
authors.initials: CQ
authors.lastname: Dai
authors.forename: Guo-Qiang
authors.initials: GQ
authors.lastname: Qian
authors.forename: Jian-Zhong
authors.initials: JZ
authors.lastname: Yu
authors.forename: Hai-Bin
authors.initials: HB
authors.lastname: He
authors.forename: Zhi-Xin
authors.initials: ZX
authors.lastname: Wang
authors.forename: Jun
authors.initials: J
authors.lastname: Hou
authors.forename: Xiao-Yin
authors.initials: XY
authors.lastname: Chen
mesh_terms: D006528:Carcinoma, Hepatocellular / Q000473:pathology* / Q000648:ultrastructure; D045744:Cell Line, Tumor; D048430:Cell Shape; D003713:Dendritic Cells / Q000187:drug effects* / Q000276:immunology*; D005936:Glucans / Q000494:pharmacology*; D006801:Humans; D016130:Immunophenotyping; D007371:Interferon-gamma / Q000378:metabolism; D018664:Interleukin-12 / Q000378:metabolism; D008113:Liver Neoplasms / Q000473:pathology* / Q000648:ultrastructure; D007959:Lymphocyte Culture Test, Mixed; D015398:Signal Transduction / Q000187:drug effects*; D013347:Subcellular Fractions / Q000187:drug effects; D051996:Transcription Factor RelA / Q000378:metabolism*
chemical_list: D005936:Glucans; D051996:Transcription Factor RelA; C568908:carboxymethylpachymaran; D018664:Interleukin-12; D007371:Interferon-gamma