Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Targeting S1P1 receptor protects against murine immunological hepatic injury through myeloid-derived suppressor cells.

doi: 10.4049/jimmunol.1301193

Figure Lengend Snippet: FIGURE 2. Treatment with FTY720 increases the liver expression of chemokines that mediate CD11b+Gr1+ myeloid cell recruitment. (A–D) C57BL/6 mice were injected with PBS or FTY720 (1 mg/kg) daily starting at 6 h before the injection of Con A (15 mg/kg). (A) At 20 h after Con A injection, the percentage of peripheral blood CD11b+Gr1+ cells was determined by flow cytometry, and the absolute numbers of CD11b+Gr1+ cells were calculated accordingly (n = 5–8). (B) CD11b+Gr1+ cells in liver were determined by flow cytometry and are presented as a representative FACS plot (left panel) and the absolute numbers of CD11b+Gr1+ cells (n = 6–8) (right panel). (C) The concentration of CXCL1 and CXCL2 in serum (left panel) or the mRNA level of CXCL1 and CXCL2 in liver (right panel) CD11b+Gr1+ cells was determined by ELISA (n = 4) or quantitative PCR (n = 4). (D) CXCR2 expression in liver CD11b+Gr1+ cells was determined by intracellular staining, followed by flow cytometry analysis (n = 3). (E and F) CIH mice were administered 50 mg anti-CXCR2 mAb (242216) or IgG isotype control (54447; both from R&D Systems) via i.v. injection. (E) Mice were sacrificed at 20 h after Con A injection, and livers were analyzed using H&E staining. Scale bar, 50 mm. Arrowheads indicate the area of necrosis. (F) The percentage of necrotic area (left panel) was quantified using ImageJ software. For each experiment, three random sections/liver were taken from four livers/group. Serum ALT levels were assessed by ELISA (n = 4) (right panel). Data are representative of four (A, B) or two (C–F) independent experiments (mean 6 SD). *p , 0.05, **p , 0.01, ***p , 0.001.

Article Snippet: The concentrations of serum CXCL1 and CXCL2 and serum or culture supernatant IFN-g were quantified by sandwich ELISA (MKC00B, MM200, and MIF00; R&D Systems).

Techniques: Expressing, Injection, Cytometry, Concentration Assay, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Staining, Control, Software

Journal: International Journal of Cancer. Journal International du Cancer

Article Title: Induction of proinflammatory mediators by CHI3L1 is reduced by chitin treatment: decreased tumor metastasis in a breast cancer model

doi: 10.1002/ijc.26379

Figure Lengend Snippet: CHI3L1 increases the expression of CCL2, CXCL2 and MMP-9. Macrophages from normal mice were cultured overnight with either 1 ng/mL or 5 ng/mL rmCHI3L1 alone (A,C,E) or in combination with LPS (1 μg/mL) (B,D,F) and cell-free supernatants were analyzed by ELISA for CCL2 (A–B) ; CXCL2 (C–D) or MMP-9 (E–F). N= 10 mice/group, (A–B) *p<0.0015, **p<0.0001, (CD)*p<0.0005, ** p<0.0002, and (E–F) *p<0.004, **p<0.0016.

Article Snippet: Cell culture supernatants and sera from control and mammary tumor bearers were analyzed for protein expression by ELISA for CCL2, CXCL2 and IFN-γ (BD Biosciences, San Jose, CA), MMP-9 and CHI3L1 (R&D Systems) according to the manufacturer's instructions.

Techniques: Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Cancer. Journal International du Cancer

Article Title: Induction of proinflammatory mediators by CHI3L1 is reduced by chitin treatment: decreased tumor metastasis in a breast cancer model

doi: 10.1002/ijc.26379

Figure Lengend Snippet: Silencing CHI3L1 decreases secretion of proinflammatory molecules by macrophages. (A) In vitro treatment of LPS (1 μg/mL) stimulated macrophages from 5-week DA-3 tumor bearers with 50 nM CHI3L1 siRNA analyzed by qRT-PCR for CHI3L1 gene expression and protein expression as determined by ELISA (B). (C–E) Purified macrophages from 5-week DA-3 tumor bearers were cultured with 50 nM CHI3L1 siRNA or 50 NM nontarget siRNA in the presence of LPS and cell-free culture supernatants analyzed for CCL2 (C), CXCL2 (D) and MMP-9 (E). N = 5 mice/group, *p < 0.05, Student's t-test

Article Snippet: Cell culture supernatants and sera from control and mammary tumor bearers were analyzed for protein expression by ELISA for CCL2, CXCL2 and IFN-γ (BD Biosciences, San Jose, CA), MMP-9 and CHI3L1 (R&D Systems) according to the manufacturer's instructions.

Techniques: In Vitro, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Purification, Cell Culture

Journal: International Journal of Cancer. Journal International du Cancer

Article Title: Induction of proinflammatory mediators by CHI3L1 is reduced by chitin treatment: decreased tumor metastasis in a breast cancer model

doi: 10.1002/ijc.26379

Figure Lengend Snippet: In vivo treatment with chitin decreases CHI3L1, CCL2, CXCL2 and MMP-9 expression in DA-3 tumor-bearing mice. (A) Serum from 2- and 5-week DA-3 tumor bearers either untreated or chitin treated (1 mg/mouse) was analyzed for CHI3L1 expression by ELISA. (B–C) Splenocytes (B) and splenic macrophages (C) from untreated or chitin treated 5-week DA-3 tumor-bearing mice were cultured overnight in the presence or absence of LPS (1 μg/mL) and cell-free supernatants were analyzed for CHI3L1 by ELISA. (D–F) Purified macrophages and T cells from spleens of 5-week DA-3 tumor-bearing mice, untreated, or chitin treated were cultured overnight with mitogens as described, and cell-free supernatants were analyzed by ELISA for CCL2 (D), CXCL2 (E) and MMP-9 (F). Data shown are the results of three independent experiments with N= 4 mice/group; *p<0.003, **p<0.002, Student's t-test

Article Snippet: Cell culture supernatants and sera from control and mammary tumor bearers were analyzed for protein expression by ELISA for CCL2, CXCL2 and IFN-γ (BD Biosciences, San Jose, CA), MMP-9 and CHI3L1 (R&D Systems) according to the manufacturer's instructions.

Techniques: In Vivo, Expressing, Enzyme-linked Immunosorbent Assay, Cell Culture, Purification