Journal:

Article Title: Adenosine mediates IL-13-induced inflammation and remodeling in the lung and interacts in an IL-13-adenosine amplification pathway

doi: 10.1172/JCI200316815

Figure Lengend Snippet: ADA enzyme therapy regulates chemokine expression in the lungs of IL-13 Tg mice. (a) MCP-3 transcript levels were quantified using real-time RT-PCR in RNA isolated from whole lungs from wild-type and Tg (IL-13 Tg) lungs at 1, 2, and 3 months of age. Values were normalized to β-actin transcript levels and are presented as the mean percentage of β-actin transcripts ± SE. Statistical significance was determined using Student t test analysis comparing wild-type to control values at each time point. n = 4 for each condition. *P = 0.005. (b) RT-PCR analysis of the levels of mRNA encoding IL-13–relevant chemokines in whole-lung RNA from 3-month-old wild-type and IL-13 Tg mice treated with PEG-ADA (+ ADA) or vehicle for 1 month. (c) Quantitative real-time RT-PCR analysis of MCP-3 transcript levels in whole-lung RNA from 3-month-old wild-type and IL-13 Tg mice treated with PEG-ADA (+ ADA) or vehicle for 1 month. Values are presented as the mean percentage of β-actin transcripts ± SE. Statistical significance was determined using Student t test analysis. *P < 0.005. n = 4 for each condition. (d) Immunohistochemistry was used to localize MCP-3 in lung sections from 3-month-old wild-type, IL-13 Tg, or IL-13 Tg mice treated with PEG-ADA for 1 month. Scale bar = 100 μm and applies to all three photographs in d.

Article Snippet: Localization was undertaken by incubating tissue sections for 30 minutes at room temperature with a 1:4 dilution of goat anti-mouse MARC (MCP-3) Ab (R&D Systems Inc.) ( 40 ) followed by development with a goat IgG Vectastain Elite ABC Kit (Vector Laboratories, Burlingame, California, USA) as described by the manufacturer.

Techniques: Expressing, Quantitative RT-PCR, Isolation, Reverse Transcription Polymerase Chain Reaction, Immunohistochemistry

Journal: Journal of Immunology Research

Article Title: Role of Lysosomes in Silica-Induced Inflammasome Activation and Inflammation in Absence of MARCO

doi: 10.1155/2014/304180

Figure Lengend Snippet: Evaluation of the effects of doses of silica and IL-1 β production in AM from WT and MARCO −/− mice. (a) Dose response to silica-stimulated IL-1 β release from AM cocultured with endotoxin (LPS at 20 ng/mL) 24 hrs was measured by ELISA. Mean ± SEM IL-1 β from cultured macrophages where filled circle ⬤ indicates MARCO −/− AM and open circle ⚪ indicates C57BL/6 WT AM. Double asterisk ** indicates P < 0.01 compared to WT control at the corresponding concentration, n = 4 per experimental group. (b) IL-1 β release from WT AM with MARCO Ab. AM were pretreated with or without MARCO Ab (5 μ g/mL) then exposed to silica (100 μ g/mL) in the presence or absence of LPS (20 ng/mL) 24 hrs. Mean ± SEM IL-1 β release from cultured macrophages was analyzed by ELISA. Triple asterisk *** indicates P < 0.001 compared to control at the corresponding treatment, n = 4 per experimental group. Unstimulated cells were used as control.

Article Snippet: For determining specificity of MARCO in IL-1 β release experiment, AM from WT mice were treated with MARCO Ab (polyclonal goat IgG, rmMARCO; aa 70–518 assession number: Q60754, Catalog number: AF2956 obtained from R&D Systems, Minneapolis, MN) 30 min prior to exposure to silica or endotoxin for 24 hr.

Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture, Control, Concentration Assay

Journal: Journal of Immunology Research

Article Title: Role of Lysosomes in Silica-Induced Inflammasome Activation and Inflammation in Absence of MARCO

doi: 10.1155/2014/304180

Figure Lengend Snippet: Effect of MARCO on NLRP3 activation using IL-1 β as an indicator and OVA as signal 1. IL-1 β release from WT and MARCO −/− AM in response to silica (100 μ g/mL) cocultured with OVA (10 mg/mL) and with or without cathepsin B inhibitor (10 μ M CA-074-Me). Mean ± SEM IL-1 β from cultured macrophages where white bars indicate WT AM and grey bars indicate MARCO −/− AM. Asterisk * indicates P < 0.05 compared to WT, n = 3 per experimental group.

Article Snippet: For determining specificity of MARCO in IL-1 β release experiment, AM from WT mice were treated with MARCO Ab (polyclonal goat IgG, rmMARCO; aa 70–518 assession number: Q60754, Catalog number: AF2956 obtained from R&D Systems, Minneapolis, MN) 30 min prior to exposure to silica or endotoxin for 24 hr.

Techniques: Activation Assay, Cell Culture

Journal: Journal of Immunology Research

Article Title: Role of Lysosomes in Silica-Induced Inflammasome Activation and Inflammation in Absence of MARCO

doi: 10.1155/2014/304180

Figure Lengend Snippet: Pro-IL-1 β mRNA expression in WT and MARCO −/− AM analyzed using RT-PCR. White bars indicate WT AM and grey bars indicate MARCO −/− AM. Fold change in pro-IL-1 β expression was compared to unstimulated control. The data represent one experiment with 2 replications.

Article Snippet: For determining specificity of MARCO in IL-1 β release experiment, AM from WT mice were treated with MARCO Ab (polyclonal goat IgG, rmMARCO; aa 70–518 assession number: Q60754, Catalog number: AF2956 obtained from R&D Systems, Minneapolis, MN) 30 min prior to exposure to silica or endotoxin for 24 hr.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Control