Journal: Frontiers in Immunology
Article Title: Harnessing the Immunomodulatory Properties of Bacterial Ghosts to Boost the Anti-mycobacterial Protective Immunity
Figure Lengend Snippet: Activation status and cytokine production of WT vs. ΔTLR4 BMMO and BMDC upon BG and LPS treatment. Uninfected and M. bovis BCG-infected BMMO and BMDC were treated with BG at MOI 40 and 5, respectively, or with a normalized amount of LPS (1,500 and 188 EU/mL, respectively) or left untreated (UT) for 24 h before analysis of surface activation markers by flow cytometry. Expression levels of CD80, CD86, CD54 for BMMO (A–C) and CD86, CD40, and CCR7 for BMDC (D–F) are shown for WT (open bar) and ΔTLR4 (black bar) cells. Cell culture supernatants were collected to quantify the levels of TNFα, IL-6, IL-10, IL12p40, and IL-12p70 produced by ELISA. Cytokine production was measured in WT and ΔTLR4 BMMO (G–J) and BMDC (K–O) . Results are expressed as mean ± SD of technical triplicates (A–F) or duplicates (G–O) and are representative of two independent experiments. Significance values were derived using 2-way ANOVA with Holm-Sidak's multiple comparisons test (ns: not significant, * p
Article Snippet: Bacteria, BG, and Mammalian Cell Culture Cultures of M. bovis BCG Pasteur strain (ATCC, 35734) were cultured in standing T25 flasks in Middlebrook 7H9 broth (Becton Dickinson Difco™, NJ, USA) at 37°C until OD600nm 0.5–0.7 and then stored at −80°C in 7H9 and 25% glycerol (1st Base, Singapore).
Techniques: Activation Assay, Infection, Flow Cytometry, Cytometry, Expressing, Cell Culture, Produced, Enzyme-linked Immunosorbent Assay, Derivative Assay