title: [Pneumococcal HSP40 induces the immune response in mouse macrophages via p38MAPK and JNK signaling pathways].
abstract: OBJECTIVE
  To investigate the mechanism of immune response in mouse macrophage induced by Pneumococcal heat shock protein 40 (HSP40).
  
  
  METHODS
  After recombinant HSP40 (rHSP40) underwent expression detection and purification, lipopolysaccharide (LPS) was removed from it. Then rHSP40 was used to stimulate bone marrow derived macrophages (BMDMs) derived from C57BL/6 wild-type mice. The mRNA levels of tumor necrosis factor α (TNF-α), interleukin 6 (IL-6), IL-1β, IL-23p19, IL-12p40, IL-12p35 and IL-10 in BMDMs were determined by reverse transcription PCR; the expressions of TNF-α, IL-6 and IL-12p40 were measured by ELISA. After stimulated by rHSP40, the levels of TNF-α and IL-6 expressed by wide-type, TLR2-/- and TLR4-/- BMDMs were detected by ELISA. The effects of the pretreatment of mitogen-activated protein kinases (MAPK) inhibitors on the secretion of TNF-α and IL-6 induced by rHSP40 were also evaluated by ELISA in BMDMs. The phosphorylation levels of p38MAPK and c-Jun N-terminal kinase (JNK) were determined by Western blotting.
  
  
  RESULTS
  The rHSP40 protein reached a purity of more than 90%. It significantly enhanced the phosphorylation levels of p38MAPK and JNK as well as the expressions of TNF-α and IL-6. The p38MAPK and JNK inhibitors significantly suppressed the expressions of TNF-α and IL-6. The expressions of TNF-α and IL-6 in TLR4-/- BMDMs significantly decreased compared with wide-type BMDMs.
  
  
  CONCLUSIONS
  HSP40-induced immune response of mouse macrophages is regulated by p38MAPK and JNK signaling pathways, and this induction process depends on TLR4.
authors.affiliation: Ministry of Education Key Laboratory of Medical Diagnostics, Department of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Yingying
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Ministry of Education Key Laboratory of Medical Diagnostics, Department of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Song
authors.initials: S
authors.lastname: Gao
authors.affiliation: Ministry of Education Key Laboratory of Medical Diagnostics, Department of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Ma
authors.affiliation: Ministry of Education Key Laboratory of Medical Diagnostics, Department of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Jingjing
authors.initials: J
authors.lastname: Cui
authors.affiliation: Ministry of Education Key Laboratory of Medical Diagnostics, Department of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Hua
authors.initials: H
authors.lastname: Yao
authors.affiliation: Ministry of Education Key Laboratory of Medical Diagnostics, Department of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Xiaoyu
authors.initials: X
authors.lastname: Sun
authors.affiliation: Ministry of Education Key Laboratory of Medical Diagnostics, Department of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Jichao
authors.initials: J
authors.lastname: Wang
authors.affiliation: Ministry of Education Key Laboratory of Medical Diagnostics, Department of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Wenchun
authors.initials: W
authors.lastname: Xu
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000276:immunology; D001854:Bone Marrow Cells / Q000166:cytology; D016207:Cytokines / Q000235:genetics; D005786:Gene Expression Regulation / Q000276:immunology; D050956:HSP40 Heat-Shock Proteins / Q000737:chemistry / Q000276:immunology*; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000302:isolation & purification; D020935:MAP Kinase Signaling System / Q000276:immunology*; D008264:Macrophages / Q000166:cytology* / Q000276:immunology* / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010766:Phosphorylation / Q000276:immunology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013296:Streptococcus pneumoniae / Q000276:immunology*; D051195:Toll-Like Receptor 2 / Q000378:metabolism; D051197:Toll-Like Receptor 4 / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D016207:Cytokines; D050956:HSP40 Heat-Shock Proteins; D008070:Lipopolysaccharides; D012333:RNA, Messenger; C493486:Tlr2 protein, mouse; C493487:Tlr4 protein, mouse; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4; D048031:JNK Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases