Journal: Mediators of Inflammation

Article Title: SP600125 Attenuates Nicotine-Related Aortic Aneurysm Formation by Inhibiting Matrix Metalloproteinase Production and CC Chemokine-Mediated Macrophage Migration

doi: 10.1155/2016/9142425

Figure Lengend Snippet: Proposed mechanism of SP600125-mediated suppression of nicotine-related AAA in the C57BL/6J mouse model. AngII, angiotensin II; JNK, c-Jun N-terminal kinase; MCP-1, monocyte chemoattractant protein-1; RANTES, regulated-on-activation, normal T-cell expressed and secreted; MMP, matrix metalloproteinase; ECM, extracellular matrix.

Article Snippet: Slides were incubated with rabbit anti-mouse RANTES immunoglobulin G (IgG), rabbit anti-mouse MCP-1 IgG, rabbit anti-mouse MMP-2, rabbit anti-mouse MMP-9, or reagent-grade nonimmune rat IgG (R&D Systems, Minneapolis, MN, USA) overnight at 4°C in a humidified chamber.

Techniques: Activation Assay

Journal: Mediators of Inflammation

Article Title: SP600125 Attenuates Nicotine-Related Aortic Aneurysm Formation by Inhibiting Matrix Metalloproteinase Production and CC Chemokine-Mediated Macrophage Migration

doi: 10.1155/2016/9142425

Figure Lengend Snippet: SP600125 suppresses the protein expression of chemokines in aortic tissue. (a) Representative transverse sections of mouse aortic tissue obtained after transient perfusion with nicotine plus AngII, with or without SP600125. MCP-1 and RANTES proteins were undetectable in normal aorta but were abundant in nicotine plus AngII-induced AAA tissues (a1 and a2), where they appear to be expressed in the medial and outer smooth muscle cells. SP600125 inhibited the protein expression of MCP-1 and RANTES in AAA lesions (a3). (c, d) Representative western blots for MCP-1 and RANTES. The band optical density (OD) values (mean ± SD) of MCP-1 and RANTES were evaluated using ImageJ. GAPDH was used as an internal control and results are from independent triplicate experiments. Chemokine expression in the serum as detected by ELISA (e). ∗∗ P < 0.01 versus coadministration.

Article Snippet: Slides were incubated with rabbit anti-mouse RANTES immunoglobulin G (IgG), rabbit anti-mouse MCP-1 IgG, rabbit anti-mouse MMP-2, rabbit anti-mouse MMP-9, or reagent-grade nonimmune rat IgG (R&D Systems, Minneapolis, MN, USA) overnight at 4°C in a humidified chamber.

Techniques: Expressing, Western Blot, Control, Enzyme-linked Immunosorbent Assay

Journal: Mediators of Inflammation

Article Title: SP600125 Attenuates Nicotine-Related Aortic Aneurysm Formation by Inhibiting Matrix Metalloproteinase Production and CC Chemokine-Mediated Macrophage Migration

doi: 10.1155/2016/9142425

Figure Lengend Snippet: Expression of MCP-1 and RANTES under a concentration gradient of nicotine in MOVAS cells. Cellular mRNA expression levels of MCP-1 and RANTES (a, b) and the levels of secreted MCP-1 and RANTES in the supernatant (c, d) are shown. MCP-1 and RANTES expression as well as secretion was induced by nicotine in a dose-dependent fashion. The strongest expression was observed for 0.5 ng/mL and 5 ng/mL nicotine. Data are from independent triplicate experiments. ∗ P < 0.05 and ∗∗ P < 0.01 versus control; # P < 0.05 and ## P < 0.01 versus the group treated with 5 ng/mL nicotine.

Article Snippet: Slides were incubated with rabbit anti-mouse RANTES immunoglobulin G (IgG), rabbit anti-mouse MCP-1 IgG, rabbit anti-mouse MMP-2, rabbit anti-mouse MMP-9, or reagent-grade nonimmune rat IgG (R&D Systems, Minneapolis, MN, USA) overnight at 4°C in a humidified chamber.

Techniques: Expressing, Concentration Assay, Control

Journal: Mediators of Inflammation

Article Title: SP600125 Attenuates Nicotine-Related Aortic Aneurysm Formation by Inhibiting Matrix Metalloproteinase Production and CC Chemokine-Mediated Macrophage Migration

doi: 10.1155/2016/9142425

Figure Lengend Snippet: Influence of SP600125 on nicotine-induced expression of MCP-1 and RANTES in MOVAS cells. Cellular mRNA expression levels of MCP-1 and RANTES (a, b) and the levels of secreted MCP-1 and RANTES in the supernatant (c, d) are shown. Nicotine at 5 ng/mL could significantly upregulate the cellular mRNA expression as well as secretion of MCP-1 and RANTES, while SP600125 eliminated this effect. Data are from independent triplicate experiments. ∗ P < 0.05 and ∗∗ P < 0.01 versus control; # P < 0.05 and ## P < 0.01 versus the group treated with 5 ng/mL nicotine.

Article Snippet: Slides were incubated with rabbit anti-mouse RANTES immunoglobulin G (IgG), rabbit anti-mouse MCP-1 IgG, rabbit anti-mouse MMP-2, rabbit anti-mouse MMP-9, or reagent-grade nonimmune rat IgG (R&D Systems, Minneapolis, MN, USA) overnight at 4°C in a humidified chamber.

Techniques: Expressing, Control

Journal: Scientific Reports

Article Title: HIV induces airway basal progenitor cells to adopt an inflammatory phenotype

doi: 10.1038/s41598-021-82143-1

Figure Lengend Snippet: HIV-induced BC expression of IL-8 mediates AM and neutrophil migration. BC were exposed to HIV or heat-inactivated HIV for 2 days. ( A ) Quantification of IL-8 in culture supernatants of untreated, heat-inactivated HIV and HIV- treated BC cultures by ELISA. ( B ) HIV-induced BC release of these mediators induce migration of human alveolar macrophages and neutrophils. Alveolar macrophages obtained from an HIV negative normal subject were plated in serum-free medium in the upper chamber and incubated with BC-conditioned media from untreated, NL4-3 treated and heat-inactivated NL4-3-treated BC culture in the lower chamber of transwell system. The number of migratory cells were quantified after 3 h. ( C ) Migration of human alveolar macrophages mediated by BC-release of IL-8. BC-conditioned media were pre-treated with mouse IgG1 control or anti-IL-8 neutralizing antibody (both at 2 µg/ml) for 30 min prior to addition to the lower chamber. Human alveolar macrophages were plated on the upper chamber and incubated with conditioned media from untreated BC, HIV-BC pretreated with mouse IgG 1 -treated or HIV-BC pretreated with anti-IL-8 neutralizing antibody. The number of migratory alveolar macrophages were counted after 3 h. Data shown are the mean ± SD of one representative of three independent experiments performed in triplicate. ( D ) Stimulation of migration of neutrophils. Human neutrophils in serum-free RPMI were plated in the upper chamber and incubated with BC-conditioned media from untreated, HIV-treated and heat-inactivated HIV-treated BC culture in the lower chamber of transwell system. The number of migratory cells were quantified after 1 h of incubation. ( E ) Migration of human neutrophils mediated by BC-released IL-8 in conditioned media. BC-conditioned media were pre-treated with mouse IgG 1 control and anti-IL-8 neutralizing antibody (both at 2 µg/ml) for 30 min prior to addition to the lower chamber. Human neutrophils were plated on the upper chamber and incubated with conditioned media from untreated BC, HIV-BC pretreated with mouse IgG 1 -treated, HIV-BC pretreated with anti-IL-8 neutralizing antibody or HIV-treated BC. The number of migratory alveolar macrophages were counted after 1 h of incubation. Data are presented as mean ± SD of one representative of three independent experiments performed in triplicate.

Article Snippet: For blocking experiments, BC-conditioned media was pre-incubated with anti-IL-8 neutralizing antibody (clone # 6217) and corresponding mouse IgG1 control (clone # 11,711; both at 2 μg/ml, endotoxin level < 0.10 EU per 1 μg of the antibody by the LAL method; R & D Systems) for 30 min at 37 oC prior to addition to the lower chamber.

Techniques: Expressing, Migration, Enzyme-linked Immunosorbent Assay, Incubation, Control

Journal: Scientific Reports

Article Title: HIV induces airway basal progenitor cells to adopt an inflammatory phenotype

doi: 10.1038/s41598-021-82143-1

Figure Lengend Snippet: HIV-treated BC release GM-CSF that stimulates AM proliferation. ( A) Quantification of GM-CSF in BC-conditioned medium by ELISA. ( B) Proliferation of human AM is partially mediated by BC-released GM-CSF in conditioned media. Alveolar macrophages in serum-free RPMI were plated in 96-well and incubated with BC-conditioned media from untreated, HIV-treated and heat-inactivated HIV-treated BC culture. RPMI and recombinant GM-CSF (20 ng/ml) serve as background and positive controls, respectively. BC-conditioned media were pre-treated with mouse IgG 1 control and anti-GM-CSF neutralizing antibody (both at 10 µg/ml) for 30 min prior to addition to the cells. Cell proliferation was measured by BrdU incorporation assay after 3 days of culture. Data represent the mean ± SD of one representative experiment (n = 3) performed in triplicate.

Article Snippet: For blocking experiments, BC-conditioned media was pre-incubated with anti-IL-8 neutralizing antibody (clone # 6217) and corresponding mouse IgG1 control (clone # 11,711; both at 2 μg/ml, endotoxin level < 0.10 EU per 1 μg of the antibody by the LAL method; R & D Systems) for 30 min at 37 oC prior to addition to the lower chamber.

Techniques: Enzyme-linked Immunosorbent Assay, Incubation, Recombinant, Control, BrdU Incorporation Assay