Journal: Infection and Immunity
Article Title: Induction of Interleukin 10 by Borrelia burgdorferi Is Regulated by the Action of CD14-Dependent p38 Mitogen-Activated Protein Kinase and cAMP-Mediated Chromatin Remodeling
Figure Lengend Snippet: CD14-dependent activation of p38 is required for IL-10 production. (A) Equal proteins from lysates of CD14 +/+ and CD14 −/− B6 BMDMs incubated with B. burgdorferi for 30 min were separated by 10% SDS-PAGE, electrotransferred to a nitrocellulose membrane, and probed with phospho-p38 and β-actin antibodies. CD14 +/+ BMDMs (B) or PMA-treated THP-1 cells (C) were treated with DMSO or increasing concentrations of SB202190 for 30 min prior to incubation with B. burgdorferi . Cell culture supernatants were collected 24 h postinfection, and TNF and IL-10 levels were measured by ELISA. (D) CD14 +/+ BMDMs were pretreated with combinations of DMSO, 5 μM SB202190, and 10, 50, or 100 ng/ml of recombinant IL-10 and then coincubated with B. burgdorferi ( B.b .) at an MOI of 10. TNF was measured in the culture supernatants collected after 24 h. (E) B. burgdorferi organisms were coincubated with PBMCs from 10 patients with Lyme arthritis, 5 with the TLR1-1805GG SNP and 5 with the TLR1-1805TG/TT SNP, for 30 min. Cell lysates were analyzed for phospho-p38 by the bead-based Luminex assay (Millipore). For panel A, data are representative of results from two independent experiments. For panels B and C, data are representative of results from five independent experiments. For panels D and E, results represent means ± SEMs from two independent experiments. ** , P
Article Snippet: BMDMs were treated with the p38 inhibitor SB202190 (each at 5 μM) (Sigma-Aldrich, St. Louis, MO) for 30 min prior to addition of B. burgdorferi at an MOI of 10 as reported earlier ( ).
Techniques: Activation Assay, Incubation, SDS Page, Cell Culture, Enzyme-linked Immunosorbent Assay, Recombinant, Luminex