title: [Use of THP-1 for allergens identification method validation].
abstract: OBJECTIVE
  Look for an in vitro test method to evaluate sensitization using THP-1 cells by the changes of the expression of cytokines to provide more reliable markers of the identification of sensitization.
  
  
  METHODS
  The monocyte-like THP-1 cells were induced and differentiated into THP-1-macrophages with PMA (0.1 microg/ml). The changes of expression of cytokines at different time points after the cells being treated with five known allergens, 2,4-dinitrochlorobenzene (DNCB), nickel sulfate (NiSO4), phenylene diamine (PPDA) potassium dichromate (K2Cr2O7) and toluene diisocyanate (TDI) and two non-allergens sodium dodecyl sulfate (SDS) and isopropanol (IPA) at various concentrations were evaluated. The IL-6 and TNF-alpha production was measured by ELISA. The secretion of IL-1beta and IL-8 was analyzed by Cytometric Bead Array (CBA).
  
  
  RESULTS
  The section of the IL-6, TNF-alpha, IL-1beta and IL-8 were the highest when THP-1 cells were exposed to NiSO4, DNCB and K2Cr2O7 for 6h, PPDA and TDI for 12h. The production of IL-6 were approximately 40, 25, 20, 50 and 50 times for five kinds chemical allergens NiSO4, DNCB, K2Cr2O7, PPDA and TDI respectively at the optimum time points and the optimal concentration compared to the control group. The expression of TNF-alpha were 20, 12, 20, 8 and 5 times more than the control group respectively. IL-1beta secretion were 30, 60, 25, 30 and 45 times respectively compared to the control group. The production of IL-8 were approximately 15, 12, 15, 12 and 7 times respectively compared to the control group. Both non-allergens SDS and IPA significantly induced IL-6 secretion in a dose-dependent manner however SDS cause a higher production levels, approximately 20 times of the control. Therefore IL-6 may not be a reliable marker for identification of allergens. TNF-alpha, IL-1beta and IL-8 expressions did not change significantly after exposed to the two non-allergens.
  
  
  CONCLUSIONS
  The test method using THP-1 cells by detecting the productions of cytokines (TNF-alpha, IL-1beta and IL-8) can effectively distinguish chemical allergens and non-allergens. The three cytokines may be reliable markers for the identification of potential sensitizing chemicals.
authors: Xuezheng
authors: X
authors: Zhao
authors: Qiang
authors: Q
authors: Jia
authors: Jun
authors: J
authors: Zhang
authors: Xue
authors: X
authors: Li
authors: Yanshu
authors: Y
authors: Zhang
authors: Yufei
authors: Y
authors: Dai
mesh_terms: D000485:Allergens; D016207:Cytokines; D004137:Dinitrochlorobenzene; D006801:Humans; D053583:Interleukin-1beta; D015850:Interleukin-6; D016209:Interleukin-8; D009000:Monocytes; D009532:Nickel; D012967:Sodium Dodecyl Sulfate; D014409:Tumor Necrosis Factor-alpha
chemical_list: D000485:Allergens; D016207:Cytokines; D004137:Dinitrochlorobenzene; D053583:Interleukin-1beta; D015850:Interleukin-6; D016209:Interleukin-8; D014409:Tumor Necrosis Factor-alpha; D012967:Sodium Dodecyl Sulfate; C029938:nickel sulfate; D009532:Nickel