title: OM85-BV induced the productions of IL-1β, IL-6, and TNF-α via TLR4- and TLR2-mediated ERK1/2/NF-κB pathway in RAW264.7 cells.
abstract: Broncho-Vaxom (OM85-BV) is an extract mixture from 8 strains of Gram(+) and Gram(-) bacteria and plays an important role in anti-infection immune response by regulating macrophage activity and cytokine productions. However, the mechanism by which OM85-BV enhances the cytokine expression is still obscure. In this study, we evaluated the effects of OM85-BV on the productions of interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α) in RAW264.7 murine macrophages. Exposure of RAW264.7 cells to 100 μg/mL OM85-BV upregulated the expression of IL-1β, IL-6, and TNF-α at the mRNA and protein levels in a time- and dose-dependent manner. In addition, OM85-BV induced extracellular signal-regulated kinase (ERK) 1/2 and nuclear factor-kappa B (NF-κB) phosphorylation. Pretreatment with U0126 or Bay11-7082, respectively, could decrease IL-1β, IL-6, and TNF-α productions induced by OM85-BV. Application of Toll-like receptor (TLR) 4 or TLR2 small-interfering RNA (siRNA) into RAW264.7 cells could inhibit the productions of cytokines and ERK1/2 and NF-κB phosphorylation induced by OM85-BV. Consistent with this, downregulating either myeloid differentiation factor 88 (MyD88) or TRIF-related adaptor molecule (TRAM) gene with MyD88-siRNA or TRAM-siRNA separately could reduce the productions of cytokines and ERK1/2 and NF-κB phosphorylation induced by OM85-BV. Our study demonstrated that the productions of IL-1β, IL-6, and TNF-α induced by OM85-BV in RAW264.7 cells were through TLR4 and TLR2 signaling pathway-mediated activation of ERK1/2 and NF-κB.
authors.affiliation: 1 Division of Nephrology, Department of Internal Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology , Wuhan, People's Republic of China .
authors.forename: Hong
authors.initials: H
authors.lastname: Luan
authors.forename: Qian
authors.initials: Q
authors.lastname: Zhang
authors.forename: Le
authors.initials: L
authors.lastname: Wang
authors.forename: Chuanxiao
authors.initials: C
authors.lastname: Wang
authors.forename: Miao
authors.initials: M
authors.lastname: Zhang
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Xu
authors.forename: Huan
authors.initials: H
authors.lastname: Zhou
authors.forename: Xing'ai
authors.initials: X
authors.lastname: Li
authors.forename: Qing
authors.initials: Q
authors.lastname: Xu
authors.forename: Fan
authors.initials: F
authors.lastname: He
authors.forename: Jin
authors.initials: J
authors.lastname: Yuan
authors.forename: Yongman
authors.initials: Y
authors.lastname: Lv
mesh_terms: D000276:Adjuvants, Immunologic / Q000494:pharmacology; D000818:Animals; D015153:Blotting, Western; D002457:Cell Extracts / Q000494:pharmacology*; D002460:Cell Line; D016207:Cytokines / Q000096:biosynthesis*; D004789:Enzyme Activation / Q000187:drug effects; D004797:Enzyme-Linked Immunosorbent Assay; D005786:Gene Expression Regulation / Q000187:drug effects*; D020935:MAP Kinase Signaling System / Q000502:physiology*; D008264:Macrophages / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D051379:Mice; D016133:Polymerase Chain Reaction; D051195:Toll-Like Receptor 2 / Q000378:metabolism*; D051197:Toll-Like Receptor 4 / Q000378:metabolism*; D000095983:Bacterial Lysates
chemical_list: D000276:Adjuvants, Immunologic; D002457:Cell Extracts; D016207:Cytokines; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4; C030259:Broncho-Vaxom; D000095983:Bacterial Lysates; C493486:Tlr2 protein, mouse; C493487:Tlr4 protein, mouse