Journal: Inflammatory bowel diseases

Article Title: Cross-talk between RORγt+ innate lymphoid cells and intestinal macrophages induces mucosal IL-22 production in Crohn's disease.

doi: 10.1097/MIB.0000000000000105

Figure Lengend Snippet: FIGURE 3. A, IL-23 secretion of sorted ILCs populations cocultured with or without macrophages, as determined by enzyme-linked immunosorbent assay (ELISA), n ¼ 3. B, The open bars show IL-22 secretion of sorted cells cocultured with macrophages under LPS stimulation measured by ELISA. The filled bars show sorted cells co- cultured with macrophages and stimulated with LPS and anti-IL-23 antibody (10 mg/mL), n ¼ 3. C, IL-23 receptor messenger RNA was quantified and normalized to 18S RNA expression, n ¼ 12. *P , 0.05.

Article Snippet: The concentration of IL-22 in cell culture supernatants of patients was assayed using a human IL-22 Quantikine enzymelinked immunosorbent assay kit (R&D Systems) with a detection limit of 15.6 pg/mL.

Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture, RNA Expression

Journal: Inflammatory bowel diseases

Article Title: Cross-talk between RORγt+ innate lymphoid cells and intestinal macrophages induces mucosal IL-22 production in Crohn's disease.

doi: 10.1097/MIB.0000000000000105

Figure Lengend Snippet: FIGURE 5. A, The percentage of each population from inflamed lesions (n ¼ 23) and noninflamed lesions (n ¼ 20). B, The open bars show IL-22 secretion levels of ILCs cocultured with macrophages from inflamed lesions. The filled bars show ILCs cocultured from noninflamed lesions in the same individuals. The secretion levels were measured by enzyme-linked immunosorbent assay, n ¼ 4. C, The open bars show IL-23 receptor expression of ILCs from inflamed lesions. The filled bars show IL-23 receptor expression of ILCs from noninflamed lesions. IL-23 receptor mes- senger RNA was quantified and normalized to 18S RNA expression, n ¼ 5. *P , 0.05. NS, not significant.

Article Snippet: The concentration of IL-22 in cell culture supernatants of patients was assayed using a human IL-22 Quantikine enzymelinked immunosorbent assay kit (R&D Systems) with a detection limit of 15.6 pg/mL.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, RNA Expression

Journal: Scientific Reports

Article Title: The protective effect of the anti-Toll-like receptor 9 antibody against acute cytokine storm caused by immunostimulatory DNA

doi: 10.1038/srep44042

Figure Lengend Snippet: ( a , c , e ) RAW264.7 cells ( a ), BM-Macs( c ) and BM-cDCs ( e ) were treated with medium, lipid A (100 ng/ml), CpGB (500 nM), IFN-β (1000 U/ml) or TNF-α (25 ng/ml). Twenty-four hours later, cells were subjected to cell surface (upper panel) or membrane-permeabilized staining (lower panel). Grey histograms show staining with isotype control. Black and red open histograms show staining with NaR9 before and after treatment. ( b , d , f ) RAW264.7 cells ( b ), BM-Macs ( d ) and BM-cDCs ( f ) were pre-treated with medium, IFN-β (1000 U/ml) or TNF-α (25 ng/ml). Twenty-four hours later, the cells were stimulated with the indicated concentration of CpGB or CpGA. Twenty-four hours after stimulation with CpGB or CpGA, culture supernatants were collected, and the concentration of RANTES ( b ) or IL-6 ( d , f ) was determined by ELISA. The results are represented by the mean value and the s.d. from triplicate wells. These experiments were repeated at least twice, and representative data are shown.

Article Snippet: ELISA Sets (eBiosciences), and IFN-α was measured using a VeriKine ELISA Kit (PBL Assay Science, Piscataway, NJ, USA).

Techniques: Membrane, Staining, Control, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: The protective effect of the anti-Toll-like receptor 9 antibody against acute cytokine storm caused by immunostimulatory DNA

doi: 10.1038/srep44042

Figure Lengend Snippet: ( a–c ) BM-Macs ( a ), BM-cDCs ( b ) and BM-pDCs ( c ) were stimulated with CpGB or CpGA. As a control, cells were stimulated by loxoribine or poly U with DOTAP. Four hours before stimulation, the cells were treated with NaR9 or isotype-matched control mAb (IgG2a) at the indicated concentrations. Supernatant was collected after 24 h of stimulation, and cytokine production was evaluated by ELISA. ( d , e ) BM-Macs ( d ) and BM-cDCs ( e ) were pre-treated with medium or IFN-β (1000 U/ml) for 24 h. The cells were treated with NaR9 or isotype-matched control mAb (IgG2a) at the indicated concentrations for 4 h and stimulated by 50 nM of CpGB. Twenty-four hours after stimulation with CpGB, culture supernatants were collected, and the concentration of IL-6 was determined by ELISA. The results are represented by the mean value and the s.d. from triplicated wells. These experiments were repeated at least 3 times ( a – c ) or twice ( d , e ), and representative data are shown.

Article Snippet: ELISA Sets (eBiosciences), and IFN-α was measured using a VeriKine ELISA Kit (PBL Assay Science, Piscataway, NJ, USA).

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

Journal: Scientific Reports

Article Title: The protective effect of the anti-Toll-like receptor 9 antibody against acute cytokine storm caused by immunostimulatory DNA

doi: 10.1038/srep44042

Figure Lengend Snippet: ( a ) C57BL/6 mice were intraperitoneally administered with anti-TLR9 or isotype-matched IgG2a mAb (25 mg/kg) 15 h before injection of CpGB (500 nmol/kg) and D-(+)-galactosamine (1 g/kg). The percentage of mice that survived the treatments is shown. Each group has 15 mice. ( b ) Blood was collected from mice at indicated time points after injection of CpGB and D-(+)-galactosamine. Serum TNF-α and IL-12p40 levels were measured by ELISA. Bars in graphs indicate the mean values. Each group has 6 mice. Statistical analysis was performed using a Student’s t-test: * p < 0.05.

Article Snippet: ELISA Sets (eBiosciences), and IFN-α was measured using a VeriKine ELISA Kit (PBL Assay Science, Piscataway, NJ, USA).

Techniques: Injection, Enzyme-linked Immunosorbent Assay