Journal: Mucosal Immunology

Article Title: PPARγ activation following apoptotic cell instillation promotes resolution of lung inflammation and fibrosis via regulation of efferocytosis and proresolving cytokines

doi: 10.1038/mi.2014.130

Figure Lengend Snippet: Peroxisome proliferator-activated receptor-γ (PPARγ) activation following apoptotic cell instillation mediates changes of transforming growth factor-β (TGF-β) and interleukin-10 (IL-10) expression over the course of bleomycin (BLM)-stimulated lung injury. Where indicated, 1 mg kg −1 GW9662 (GW) was administered intraperitoneally (i.p.) at the same time with apoptotic cell (ApoJ) instillation 2 days after BLM treatment and every day thereafter. ( a–c , g–i ) Mice were killed at 2 h after ApoJ instillation and on ( d – f , j – l ) days 7, 14, and 21 after BLM treatment. Sal, saline alone. TGF-β and IL-10 mRNA levels in ( a , d , g and j ) alveolar macrophages (AM) and ( b , e , h and k ) lung homogenates were analyzed by real-time PCR. ( c , f , i and l ) TGF-β1 and IL-10 protein levels in bronchoalveolar lavage fluid (BALF) were quantified by enzyme-linked immunosorbent assay (ELISA). Values represent the mean±s.e.m. from ( a , b , d , e , g , h , j and k ) 5 or ( c , f , i and l ) 10 mice in each group. + P <0.05 for BLM+ApoJ vs. BLM+Sal, # P <0.05 for BLM+ApoJ vs. BLM+ApoJ+GW. * P <0.05 compared with saline control.

Article Snippet: BAL fluid samples were assayed using tumor necrosis factor-α, macrophage inflammatory protein-2, IL-4, IL-13, IFN-γ, TGF-β1, IL-10, and HGF ELISA kits as per the manufacturer's instructions (R&D Systems, Minneapolis, MN).

Techniques: Activation Assay, Expressing, Saline, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Mucosal Immunology

Article Title: PPARγ activation following apoptotic cell instillation promotes resolution of lung inflammation and fibrosis via regulation of efferocytosis and proresolving cytokines

doi: 10.1038/mi.2014.130

Figure Lengend Snippet: A mechanism diagram summarizing and integrating the effects of enhanced peroxisome proliferator-activated receptor-γ (PPARγ) activity following apoptotic cell instillation in bleomycin-induced lung fibrosis. Arg1, arginase 1; Col1, type 1 collagen α2; ECM, extracellular matrix; Fn, fibronectin; HGF, hepatocyte growth factor; IL-10, interleukin-10; MIP-2, macrophage inflammatory protein-2; MMR, macrophage mannose receptor; α-SMA, α-smooth muscle actin; TGF-β, transforming growth factor-β TNF-α, tumor necrosis factor-α Th1/Th2, helper T cell type 1/2.

Article Snippet: BAL fluid samples were assayed using tumor necrosis factor-α, macrophage inflammatory protein-2, IL-4, IL-13, IFN-γ, TGF-β1, IL-10, and HGF ELISA kits as per the manufacturer's instructions (R&D Systems, Minneapolis, MN).

Techniques: Activity Assay

Journal:

Article Title: Taxane-mediated gene induction is independent of microtubule stabilization: Induction of transcription regulators and enzymes that modulate inflammation and apoptosis

doi:

Figure Lengend Snippet: Quantitative analysis of TNFα formation and caspase-3 activity as a function of time in RAW 264.7 cells incubated with taxol (▪) or taxotere (•). In the media from cells incubated with taxol, TNFα levels increased by 4 hr and remained constant between 4 and 24 hr, consistent with the temporal profile of mRNA expression in Fig. ​Fig.22A. TNFα was undetectable from 0 to 24 hr in media from cells incubated with taxotere, consistent with data in Fig. ​Fig.22A. From 24 to 48 hr, TNFα levels in the media increased, modestly, for cells incubated with taxol and taxotere (A). IL-10 inhibited the formation of TNFα by cells incubated with taxol (B). Caspase-3 activity increased as a function of time in RAW 264.7 cells incubated with taxol (▪) or taxotere (•) compared with vehicle control (▴) (C). Increased caspase-3 activity at 24–36 hr reflects the initiation of apoptosis in cells incubated with either taxol or taxotere. Caspase-3 activity continued to increase between 36 and 48 hr only in cells incubated with taxol, reflecting incrementally increased apoptosis compared with cells incubated with taxotere (C). IL-10, which inhibited TNFα formation, also inhibited casapase-3 activity in cells treated with taxol. Caspase-3 activity in cells treated with taxol plus IL-10 was indistinguishable from the caspase-3 activity in cells treated with taxotere (D).

Article Snippet: In certain experiments we quantified caspase activity in cells incubated for 48 hr with combinations of: ( i ) 10 ng/ml murine recombinant interleukin IL-10 (R & D Systems) plus 10 μM taxol, ( ii ) 2.5 ng/ml murine recombinant TNFα (R & D Systems) plus 10 μM taxotere, or ( iii ) 20 μl TNFα neutralizing polyclonal antibody (Genzyme) plus 10 μM taxol.

Techniques: Activity Assay, Incubation, Expressing

Journal: British Journal of Cancer

Article Title: Impaired access of lymphocytes to neoplastic prostate tissue is associated with neoangiogenesis in the tumour site

doi: 10.1038/sj.bjc.6603650

Figure Lengend Snippet: Expression of IL-10 in adenocarcinoma lesions (PAC), adjacent benign tissue (peri-PAC) and specimens of nodular hyperplasia of the prostate gland (NHPG). ( A ) Typical pattern of IL-10 expression in malignant and benign prostate tissue (magnification × 200). ( B ) Percentages of IL-10-expressing specimens.

Article Snippet: Sections of prostate tissue were reacted to commercial antibodies diluted, as follows: monoclonal anti-CD34 (Dako Cytomation A/S, Glosstrop, Denmark), diluted 1 : 100; monoclonal anti-CD54 (Zymed Laboratories Inc., South San Francisco, CA, USA) ready to use; monoclonal anti-CD106 (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA), diluted 1 : 25 and 1 : 50; monoclonal anti-CD106 (Dako Cytomation), diluted 1 : 25; monoclonal CD62E (Santa Cruz), diluted 1 : 25; monoclonal antiinterleukin (IL)-10 (R&D Systems Europe Ltd, Abington, UK), diluted 1 : 20; monoclonal anti-CD45 (Dako Cytomation), diluted 1 : 100; anti-CD3 (Dako Cytomation), diluted 1 : 100; anti-CD4 (Zymed) ready to use; anti-CD8 (Dako Cytomation), diluted 1 : 25; anti-CD20 (Dako Cytomation), diluted 1:100; anti-CD14 (Zymed), diluted 1 : 50; and anti-CD15 (Dako Cytomation), diluted 1 : 50.

Techniques: Expressing