anti tgf β1 neutralizing antibody  (Proteintech)

Journal: JCI insight

Article Title: TGF-β1 induces PD-1 expression in macrophages through SMAD3/STAT3 cooperative signaling in chronic inflammation.

doi: 10.1172/jci.insight.165544

Figure Lengend Snippet: Figure 2. TGF-β1 is involved in PD-1 expression on macrophages in chronic inflammatory tissues. (A) Correlations between PDCD1 and TGFB1 gene expression in various human tumors by TIMER web server (TIMER2.0; https://cistrome.shinyapps.io/timer/). Abbreviations for various human tumors are given according to the database. COAD, colon adenocarcinoma. (B) Correlation plot between PDCD1 and TGFB1 gene levels in COAD (n = 458; Spearman correlation coefficient = 0.593, P = 9.38 × 10–45). (C and D) Immunoblot analysis of TGF-β1 expression levels in the MC38 tumors and normal colon tissues of tumor-bearing mice (C) or in the livers of normal uninfected and S. japonicum–infected mice (8 weeks postinfection; D). TGF-β1 expression levels were normalized to GAPDH. (E and F) Recombinant TGF-β1, PBS, anti–TGF-β1 neutralizing antibody, or isotype control antibody was administrated intraperito- neally into MC38 tumor–bearing (E) or S. japonicum–infected mice (F). PD-1+ macrophages in the tumor tissue or liver were analyzed using flow cytometry. Representative histograms and quantification of PD-1+ macrophages are shown. Spearman’s rank correlation coefficient (A and B) or an unpaired 2-tailed t test (C–F) was used for statistical analysis. The data are expressed as the mean ± SD of 3–5 mice per group and are representative of 2 independent experiments. **P < 0.01, ***P < 0.001. TPM, transcripts per million.

Article Snippet: Anti–TGF-β1 neutralizing antibody (BE0057; BioXCell; 250 μg per mouse) was administrated intraperitoneally into mice daily every 3 days, starting at 6 weeks postinfection until 8 weeks postinfection or starting at 7 days after tumor implantation until 21 days postimplantation.

Techniques: Expressing, Gene Expression, Western Blot, Infection, Recombinant, Control, Flow Cytometry

Journal: Cells

Article Title: Tumor-Induced T Cell Polarization by Schwann Cells

doi: 10.3390/cells11223541

Figure Lengend Snippet: Tumor-derived TGF-β induced PGE2 production by Schwann cells. Adult SC were isolated from the sciatic nerve of C57BL/6 mice, cultured and purified as described in M&M. TGF-β production by melanoma B16, ret and BPWT cells and SC were assessed by ELISA in cell-free supernatant obtained from tumor cells and SC after they were washed and re-cultured for 24 h ( A ). SC were also co-cultured with medium (SC), B16-conditioned medium (10% v / v ) (SC/B16) and 10 ng/mL TGF-β1 (SC/TGF-b) for 24 h, washed and cultured for additional 24 h. PGE2 levels were then determined in cell-free supernatants by ELISA ( B ). In addition, SC were cultured with medium (SC), B16-conditioned medium (10% v / v ) (SC/B16) and conditioned medium from B16 cells that were cultured in the presence of anti-TGF-β neutralizing antibody (SC/B1b+aTFGb). Control and tumor-treated SC were then washed and cultured for another 24 h to assess PGE2 production by ELISA ( C ). Results are the mean ± SEM. *, p < 0.001 versus control SC group (n = 3, One-way ANOVA).

Article Snippet: Medium was removed and SC were treated with tumor-conditioned medium (10% v / v ), control medium or r TGF-β1 (100 pg/mL, Cell Signaling Technology, Danvers, MA, USA) for 24 h. To block the TGF-β1 activity, 10 μg/mL anti-TGF-β1-neutralizing antibody (R&D System, Minneapolis, MN, USA) was added at the beginning of stimulation.

Techniques: Derivative Assay, Isolation, Cell Culture, Purification, Enzyme-linked Immunosorbent Assay

Journal: Cells

Article Title: Tumor-Induced T Cell Polarization by Schwann Cells

doi: 10.3390/cells11223541

Figure Lengend Snippet: Tumor activated SC for PGE2 production via SMAD2/3 and ERK1/2 pathways. Adult SC were isolated from the sciatic nerve of C57BL/6 mice, cultured and purified as described in M&M. SC were then cultured with B16-conditioned medium (10% v / v ) for 0–180 min, harvested, washed in cold medium and used for cell lysate preparation for protein expression analyses. Expression of phosphorylated (p) and total (t) SMAD2/3 and ERK1/2 proteins in control and tumor-treated SC was determined by Western blot analysis after proteins were separated by SDS-PAGE electrophoresis and immunoblotted as described in M&M. Representative Western blot results are shown ( A ). SC were also co-cultured with medium (SC), B16-conditioned medium (10% v / v ) (SC/B16) and 10 ng/mL TGF-β1 (SC/TGF) for 24 h after pre-treatment with either 10 µM SB431542, a selective inhibitor of the TGF-βRI/ALK4/5/7 (SB) or 10 µM UO126, a selective inhibitor of MAPK/ERK kinase (UO). Pre-treatment with medium served as a control. Cells were then washed and cultured for additional 24 h. PGE2 levels were then determined in cell-free supernatants by ELISA ( B ). Results are the mean ± SEM. *, p < 0.001 versus control SC group; #, p < 0.001 versus B16- and TGF-β-treated SC groups, respectively (n = 3, One-way ANOVA).

Article Snippet: Medium was removed and SC were treated with tumor-conditioned medium (10% v / v ), control medium or r TGF-β1 (100 pg/mL, Cell Signaling Technology, Danvers, MA, USA) for 24 h. To block the TGF-β1 activity, 10 μg/mL anti-TGF-β1-neutralizing antibody (R&D System, Minneapolis, MN, USA) was added at the beginning of stimulation.

Techniques: Isolation, Cell Culture, Purification, Expressing, Western Blot, SDS Page, Electrophoresis, Enzyme-linked Immunosorbent Assay