Journal: Gut

Article Title: SR-PSOX/CXCL16 plays a critical role in the progression of colonic inflammation.

doi: 10.1136/gut.2010.221879

Figure Lengend Snippet: Figure 2 SR-PSOX/CXCL16 plays a role in phagocytosis of bacterial components and the production of interleukin 12 (IL-12) by macrophages. Thioglycollate-elicited peritoneal macrophages from SR-PSOX/CXCL16 knockout (KO) and wild-type (WT) mice were subjected to an ex vivo phagocytosis assay against bacteria. (A) Microscopic observation of phagocytosis was performed in peritoneal macrophages from WT mice (upper column) and SR-PSOX/CXCL16 KO mice (lower column). The upper and lower panels of each column show light microscopic findings and fluorescent images, respectively. Scale bars, 25 mm. (B) Fluorescence intensity, from which the fluorescence values of the no-cell background control wells were subtracted, was measured using a microplate reader at the indicated times. The statistical comparison of fluorescence intensity between WT (open circles) and SR-PSOX/CXCL16 KO macrophages (filled circles) was assessed by repeated measure analysis of variance followed by unpaired Student t test. (C) Peritoneal macrophages from WT (open bars) and SR-PSOX/CXCL16 KO mice (filled bars) were incubated with 500 U/ml interferon g (IFNg) for 16 h, followed by stimulation with 100 ng/ml lipopolysaccharide (LPS) or 30 mg caecal bacterial lysate (CBL) for 24 h, and the culture supernatants were analysed by ELISA to measure the concentrations of IL-6 and IL-12/23 p40. The results are expressed as means6SEM of the data from three independent experiments. The statistical difference was determined by unpaired Student t test. *p<0.05 and **p<0.01 between SR-PSOX/ CXCL16 KO and WT macrophages.

Article Snippet: For SR-PSOX/CXCL16 immunostaining, colonic tissues of WT mice with or without DSS-induced colitis, and Peyer ’s patches as the positive control, were prepared as described previously.24 The sectionswere incubatedwith biotinylated anti-CXCL16 antibody (1:250; R&D Systems) or goat IgG isotype control overnight at 48C.

Techniques: Knock-Out, Ex Vivo, Phagocytosis Assay, Bacteria, Fluorescence, Control, Comparison, Incubation, Enzyme-linked Immunosorbent Assay

Journal: Gut

Article Title: SR-PSOX/CXCL16 plays a critical role in the progression of colonic inflammation.

doi: 10.1136/gut.2010.221879

Figure Lengend Snippet: Figure 3 SR-PSOX/CXCL16 levels are higher in mice with dextran sulfate sodium (DSS)-induced colitis. (A) The serum levels of SR-PSOX/ CXCL16 in mice with DSS-induced colitis on day 8 and control mice were measured by ELISA. (B). The gene expression of SR-PSOX/CXCL16 in colonic tissues with or without DSS-induced colitis was determined by quantitative real-time reverse transcriptionePCR (RTePCR) and was normalised to glyceraldehyde phosphate dehydrogenase (GAPDH). (C). The production of SR-PSOX/CXCL16 in colonic tissues with or without DSS-induced colitis was investigated by western blot analysis. (D) SR- PSOX/CXCL16 concentrations in supernatants of colon fragment cultures were measured by ELISA. The results are expressed as means6SEM (n¼10 in each group). (A), (B) and (D) The statistical difference was determined by unpaired Student t test. *p<0.05 and **p<0.01 between mice with DSS-induced colitis and normal controls.

Article Snippet: For SR-PSOX/CXCL16 immunostaining, colonic tissues of WT mice with or without DSS-induced colitis, and Peyer ’s patches as the positive control, were prepared as described previously.24 The sectionswere incubatedwith biotinylated anti-CXCL16 antibody (1:250; R&D Systems) or goat IgG isotype control overnight at 48C.

Techniques: Control, Enzyme-linked Immunosorbent Assay, Gene Expression, Western Blot

Journal: Gut

Article Title: SR-PSOX/CXCL16 plays a critical role in the progression of colonic inflammation.

doi: 10.1136/gut.2010.221879

Figure Lengend Snippet: Figure 4 SR-PSOX/CXCL16 is expressed predominantly on macrophages in colonic tissues of mice with dextran sulfate sodium (DSS)-induced colitis. (A) Immunostaining was performed in Peyer’s patches as a positive control, normal colons and colons with 3% DSS-induced colitis. Serial sections of each tissue were stained with H&E, control goat immunoglobulin G (IgG) and anti-mouse SR-PSOX/CXCL16 monoclonal antibody. (B). Immunofluorescent staining was performed in normal colons and colons with 3% DSS-induced colitis using antibodies against CD11b (green), SR-PSOX/CXCL16 (red) and control goat IgG. The merged images and their magnified images are shown. Scale bars, 50 mm (A), 20 mm (B, left 3 lanes) and 10 mm (B, right lane).

Article Snippet: For SR-PSOX/CXCL16 immunostaining, colonic tissues of WT mice with or without DSS-induced colitis, and Peyer ’s patches as the positive control, were prepared as described previously.24 The sectionswere incubatedwith biotinylated anti-CXCL16 antibody (1:250; R&D Systems) or goat IgG isotype control overnight at 48C.

Techniques: Immunostaining, Positive Control, Staining, Control

Journal: Gut

Article Title: SR-PSOX/CXCL16 plays a critical role in the progression of colonic inflammation.

doi: 10.1136/gut.2010.221879

Figure Lengend Snippet: Figure 5 Activity of dextran sulfate sodium (DSS)-induced colitis is lower in SR-PSOX/CXCL16 knockout (KO) mice. (A) Serial change in body weight in SR-PSOX/CXCL16 KO and wild-type (WT) mice with or without 3% DSS-induced colitis. Data are expressed as the percentage change from the starting body weight. (B) Representative image and colonic length in SR-PSOX/CXCL16 KO and WT mice with or without 3% DSS-induced colitis on day 8. (C) Representative histological findings and the scores of colonic inflammation of SR-PSOX/CXCL16 KO and WT mice withor without 3% DSS-induced colitis on day 8. Scale bars, 100 mm. (D) Colonic tissues of SR-PSOX/CXCL16 KO and WT mice with or without 3% DSS-induced colitis were incubated with fluorescein isothiocyanate (FITC)-conjugated CD3, CD11b and CD11c, followed by nuclear counterstaining with 4‘,6-diamidino-2-phenylindole (DAPI). Scale bars, 100 mm. (E) Fluorescent in situ hybridization analysis was performed with colonic tissues of SR-PSOX/CXCL16 KO and WT mice with or without 3% DSS-induced colitis using eubacterial oligonucleotide probe EUB-338 (red), followed by DAPI (blue). Scale bars, 50 mm (left two lanes) and 20 mm (right lane). (F) Colonic macrophages from SR-PSOX/CXCL16 KO and WT mice were subjected to an ex vivo phagocytosis assay against bacteria. Scale bars, 25 mm. (G) MLN cells from SR-PSOX/CXCL16 KO and WT mice on days 0, 5 and 8 after administration of 3% DSS were cultured with immobilised anti-CD3 plus CD28. Supernatants were collected after 72 h and subjected to ELISA to measure the concentration of interferon g (IFNg) and interleukin 17 (IL-17). (A)e(D), (F) and (G) The results are expressed as means6SEM (n¼10e12 in each group). The statistical comparison was assessed by repeated measure analysis of variance followed by unpaired Student t test (A) and (F). The statistical difference was determined by unpaired Student t test (B), (D) and (G) or ManneWhitney U test (C). *p<0.05 and **p<0.01 between SR-PSOX/CXCL16 KO mice and WT mice with DSS-induced colitis.

Article Snippet: For SR-PSOX/CXCL16 immunostaining, colonic tissues of WT mice with or without DSS-induced colitis, and Peyer ’s patches as the positive control, were prepared as described previously.24 The sectionswere incubatedwith biotinylated anti-CXCL16 antibody (1:250; R&D Systems) or goat IgG isotype control overnight at 48C.

Techniques: Activity Assay, Knock-Out, Incubation, In Situ Hybridization, Ex Vivo, Phagocytosis Assay, Bacteria, Cell Culture, Enzyme-linked Immunosorbent Assay, Concentration Assay, Comparison

Journal: Parasitology international

Article Title: Short-term protection conferred by Leishvacin® against experimental Leishmania amazonensis infection in C57BL/6 mice.

doi: 10.1016/j.parint.2014.07.010

Figure Lengend Snippet: Fig. 2. IgG1 (A) and IgG2a (B) binding to L. amazonensis were measured by ELISA, as de- scribed in Materials and methods. Serum from control and vaccinated C57BL/6 mice were collected at 4, 8 and 16 weeks post-infection. Data are representative of three exper- iments. Results for week 16 are from a different experiment than the ones for weeks 4 and 8. *P b 0.05.

Article Snippet: Mouse sera were diluted 1:10 with PBS-5% FBS and incubatedwith goat anti-mouse IgG1 or IgG2a (Southern Biotechnology, Birmingham, AL, USA) at a 1:5000 and 1:10000 dilution.

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Control, Infection

Journal: Parasitology international

Article Title: Short-term protection conferred by Leishvacin® against experimental Leishmania amazonensis infection in C57BL/6 mice.

doi: 10.1016/j.parint.2014.07.010

Figure Lengend Snippet: Fig. 5. Course of infection in vaccinated and control IFN-γ−/−(A) and iNOS−/−(C) mice infected with 105 L. amazonensis in the right hind footpad. Each point represents the difference in size ± standard deviation of the mean between infected and uninfected footpads for five mice per group. Parasite burdens were determined at 16 weeks post-infection at the site of infection in vaccinated and non-vaccinated IFN-γ−/−(B) and iNOS−/−(D) mice (5 mice per group). Levels of anti-L. amazonensis IgG1 (E) and IgG2a (F) were measured by ELISA, as described in Materials and Methods. Data are representative of three or more experiments. *P b 0.05 indicates a difference between control and vaccinated IFN-γ−/−mice and #P b 0.05 indicates difference between control and vaccinated iNOS−/−mice.

Article Snippet: Mouse sera were diluted 1:10 with PBS-5% FBS and incubatedwith goat anti-mouse IgG1 or IgG2a (Southern Biotechnology, Birmingham, AL, USA) at a 1:5000 and 1:10000 dilution.

Techniques: Infection, Control, Standard Deviation, Enzyme-linked Immunosorbent Assay