title: [Effect of monocytes activated with lipopolysaccharide on human Th17 cell differentiation].
abstract: OBJECTIVE
  To investigate whether monocytes activated with lipopolysaccharide(LPS) have an effect on Th17 cell differentiation in humans, CD4(+) T cell and CD14(+) monocytes activated with LPS were treated in the absence or presence of anti-CD3 mAb with various concentrations at different time points.
  
  
  METHODS
  Purification of CD4(+) T cell and CD14(+) monocytes were performed by magnetic cell sorting and cultured together. Cultures were stimulated with LPS alone or anti-CD3 mAb alone or LPS plus anti-CD3 mAb for 3 days. In the anti-CD3 mAb stimulation cells were added different concentrations of LPS. Cells were activated under LPS/anti-CD3 costimulation for 3, 6, or 10 days. The percentage of IL-17(+) T cells and INF-γ(+) T was determined by flow cytometry.
  
  
  RESULTS
  LPS or anti-CD3 mAb alone induced only very low levels of IL-17(+) T cells, (1.30 ± 0.19)%, (1.10 ± 0.21)%, respectively. The percentage was substantially higher in the LPS and anti-CD3 mAb costimulationa as much as(2.01 ± 0.46)%. In the presence of 0.1 μg/mL, 1 μg/mL, 10 μg/mL LPS, the proportion of Th17 reached to (1.92 ± 0.21)%, (1.30 ± 0.37)%, (1.01 ± 0.25)%. Low-concentration LPS (0.1 μg/mL) stimulation favored Th17 differentiation. The highest proportion of IL-17(+) T cells was found at day 3(2.13 ± 0.32)%, with levels declining at day 6 and day 10, while, Th1 at day 6(17.45 ± 3.04)%, declining at day 10.
  
  
  CONCLUSIONS
  Low-concentration LPS stimulation plus anti-CD3 mAb in short term support optimal Th17 generation. Nevertheless, this model closely mimics the environment of rheumatoid arthritis in vivo and proposes an effective model for the generation of human Th17 cells.
authors.affiliation: Department of Clinical Immunology,  Xijing Hospital,  Fourth Military Medical  University,  Xi'an 710032,  China.
authors.forename: Hui
authors.initials: H
authors.lastname: Yang
authors.forename: Ying
authors.initials: Y
authors.lastname: Ren
authors.forename: Ping
authors.initials: P
authors.lastname: Zhu
authors.forename: Li-Na
authors.initials: LN
authors.lastname: Chen
authors.forename: Ming-Hua
authors.initials: MH
authors.lastname: Lv
authors.forename: Chun-Mei
authors.initials: CM
authors.lastname: Fan
authors.forename: Yan-Hong
authors.initials: YH
authors.lastname: Wang
mesh_terms: D000911:Antibodies, Monoclonal / Q000494:pharmacology; D017252:CD3 Complex / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects / Q000276:immunology*; D002478:Cells, Cultured; D018920:Coculture Techniques; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry; D006801:Humans; D007371:Interferon-gamma / Q000276:immunology / Q000378:metabolism; D020381:Interleukin-17 / Q000276:immunology / Q000378:metabolism; D018950:Lipopolysaccharide Receptors / Q000276:immunology / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D008213:Lymphocyte Activation / Q000187:drug effects / Q000276:immunology; D009000:Monocytes / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D058504:Th17 Cells / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D013997:Time Factors
chemical_list: D000911:Antibodies, Monoclonal; D017252:CD3 Complex; D020381:Interleukin-17; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D007371:Interferon-gamma