title: [Bioimmunological characteristics of mature or immature murine dendritic cells].
abstract: OBJECTIVE
  To explore the biological characteristics of immature and mature murine bone marrow-derived dendritic cells (DCs).
  
  
  METHODS
  The murine bone marrow cells were cultured and induced in vitro into immature DCs (imDCs). Then ImDCs were incubated with TNF (tumor necrosis factor)-α to obtain mature DCs (mDCs). The DCs were purified by murine CD11c microbeads. The morphologies of DCs were observed by electron microscopy (EM). Such surface markers as MHC-II, CD80 and CD86 were tested by flow cytometry (FCM). The proliferation capacity of allogeneic T cells stimulated by DCs was examined by Cell Counting Kit-8 (CCK-8) in mixed lymphocyte reaction (MLR). The expressions of cell factors in mRNA of DCs were tested by real-time polymerase chain reaction (QPCR). The cytokines in supernatant were measured by ELISA (enzyme-linked immunosorbent assay).
  
  
  RESULTS
  As compared with mDCs, fewer, shorter spines and more phagocytic vesicles and lysosome were observed in imDCs under EM. The expressions of cell surface molecules in imDCs were significantly lower than those of mDC by FCM, [MHC-II (27.2%) vs (97.7%); CD80 (27.6%) vs (97.2%); CD86 (29.5%) vs (96.4%)]. In MLR, the capacity of same-reaction ratio imDCs group for stimulating the proliferation of T-cells was remarkably lower than that of mDCs group,[1:5 (1.63 ± 0.04) vs (2.21 ± 0.09); 1:10 (1.50 ± 0.08) vs (1.92 ± 0.02); 1:20 (1.28 ± 0.07) vs (1.64 ± 0.01); 1:40 (1.19 ± 0.04) vs (1.45 ± 0.06), P < 0.01]. In imDCs, the relative mRNA expressions of IL12p35 (0.66 ± 0.13), IL12p40 (0.57 ± 0.10) and IFN-γ (0.74 ± 0.08) were lower than those of mDCs (1.00 ± 0.00), (P < 0.05), but the expression of TGF-β (1.35 ± 0.09) was higher than that of mDCs by QPCR (1.00 ± 0.00), (P < 0.05). The expressions of IL12p70 and IFN-γ in supernatants from imDCs were lower than those of mDCs [IL12p70: (6 ± 4) vs (120 ± 22); IFN-γ: (56 ± 15) vs (90 ± 15), P < 0.05] while the expression of TGF (transforming growth factor)-β was higher than that of mDCs by ELISA [TGF-β: (176 ± 23) vs (55 ± 18), P < 0.05].
  
  
  CONCLUSIONS
  ImDCs can induce anergic T cells and immune tolerance through a down-regulation of co-stimulatory molecules, MHC-II and Th1 cytokines and an up-regulation of Th2 cytokine (TGF-β). And MDCs may activate naive T cells and stimulate immune responses by an up-regulation of co-stimulatory molecules, MHC-II and Th1 cytokines.
authors.affiliation: Department of Neurology, Second Hospital of Hebei Medical University, Shijiazhuang, China.
authors.forename: Ke-ling
authors.initials: KL
authors.lastname: Wang
authors.forename: Li
authors.initials: L
authors.lastname: Guo
authors.forename: Rong-fu
authors.initials: RF
authors.lastname: Shi
authors.forename: Jin-li
authors.initials: JL
authors.lastname: Wang
authors.forename: Shi-hao
authors.initials: SH
authors.lastname: Dong
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology / Q000378:metabolism*; D007371:Interferon-gamma / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D016212:Transforming Growth Factor beta / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D016212:Transforming Growth Factor beta; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma