title: [Blocking p38MAPK pathway inhibits the proliferation of OT-II cells mediated by splenic dendritic cells].
abstract: OBJECTIVE
  To investigate the effect of blocking p38 mitogen-activated protein kinase (p38MAPK) pathway on the proliferation of OT-II cells mediated by splenic dendritic cells (DCs).
  
  
  METHODS
  Splenic DCs of C57BL/6 mice were purified with anti-CD11c immunomagnetic beads, and OT-II cells were isolated from the splenic of CD4(+)-ovalbumin transgenic mice (OT-IItransgenic mice) by mouse CD4 T cell isolation kits. After being pretreated with SB203580, an inhibitor of p38MAPK, DCs were stimulated with lipopolysaccharides (LPS). Then the expression levels of co-stimulatory molecules (CD80, CD86) and MHCII in DCs, and antigen-presenting ability of DCs treated with the 52-68 fragment of the E alpha-chain of I-E class II molecules (Eα52-68 peptide) were detected by flow cytometry. The protein levels of tumor necrosis factor α (TNF-α), interleukin 1α (IL-1α), interleukin 6 (IL-6) and transforming growth factor β (TGF-β) in the culture supernatants were measured by ELISA. The proliferation of OT-II cells which were co-cultured with OVA323-339-treated DCs was analyzed by flow cytometry.
  
  
  RESULTS
  The purity of both DCs and OT-II cells reached over 90% after isolation. SB203580 downregulated the expressions of CD80, CD86 and MHC II, and suppressed the antigen-presenting ability of DCs. The expressions of TNF-α, IL-1α and IL-6 were downregulated, while the expression of TGF-β was raised. Finally, SB203580 inhibited DCs-mediated proliferation of OT-II cells.
  
  
  CONCLUSIONS
  Blocking p38MAPK pathway with SB203580 could inhibit DCs-mediated proliferation of OT-II cells, which might be involved in modulating the expressions of CD80, CD86 and MHC II, the antigen-presenting ability, as well as the expressions of pro-inflammatory and anti-inflammatory cytokines.
authors.affiliation: Department of Biotechnology, Jinan University, Guangzhou 510632, China.
authors.forename: Ling
authors.initials: L
authors.lastname: Han
authors.affiliation: State Key Laboratory of Hematology, Hospital of Blood Disease, Chinese Academy of Medical Sciences, Tianjin 300020, China.
authors.forename: Yuechen
authors.initials: Y
authors.lastname: Luo
authors.affiliation: State Key Laboratory of Hematology, Hospital of Blood Disease, Chinese Academy of Medical Sciences, Tianjin 300020, China.
authors.forename: Wantong
authors.initials: W
authors.lastname: Wu
authors.affiliation: Department of Biotechnology, Jinan University, Guangzhou 510632, China. *Corresponding authors, E-mail: wyfmsm@163.com.
authors.forename: Yongfei
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Biotechnology, Jinan University, Guangzhou 510632, China. *Corresponding authors, E-mail:msmwdw@163.com.
authors.forename: Sanmei
authors.initials: S
authors.lastname: Ma
mesh_terms: D000818:Animals; D017951:Antigen Presentation / Q000187:drug effects / Q000276:immunology; D018122:B7-1 Antigen / Q000378:metabolism; D051940:B7-2 Antigen / Q000378:metabolism; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D049109:Cell Proliferation / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D018920:Coculture Techniques; D016207:Cytokines / Q000378:metabolism; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005434:Flow Cytometry; D000949:Histocompatibility Antigens Class II / Q000378:metabolism; D007093:Imidazoles / Q000494:pharmacology; D018836:Inflammation Mediators / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D020935:MAP Kinase Signaling System / Q000187:drug effects / Q000502:physiology*; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D010446:Peptide Fragments / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D011948:Receptors, Antigen, T-Cell; D013154:Spleen / Q000166:cytology / Q000276:immunology; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: D018122:B7-1 Antigen; D051940:B7-2 Antigen; D016207:Cytokines; D004791:Enzyme Inhibitors; D000949:Histocompatibility Antigens Class II; D007093:Imidazoles; D018836:Inflammation Mediators; D008070:Lipopolysaccharides; D010446:Peptide Fragments; D011725:Pyridines; D011948:Receptors, Antigen, T-Cell; C117647:antigenic peptide Ealpha52-68; D048051:p38 Mitogen-Activated Protein Kinases; C093642:SB 203580