Journal: The Journal of Experimental Medicine

Article Title: Small intestine lamina propria dendritic cells promote de novo generation of Foxp3 T reg cells via retinoic acid

doi: 10.1084/jem.20070602

Figure Lengend Snippet: Intestinal DCs induce Foxp3 expression of effector T cells with higher efficiency than splenic DC. (A) 10 5 Foxp3 − cells were cultured with 2 × 10 4 splenic or LpDCs for 5 d with α-CD3 mAb alone or combined with TGF-β. Plots are gated on CD4 + cells, and the percentages of Foxp3 + cells are shown. Phenotypic analysis of converted T reg cells is shown on the right. 5-d cultured cells from A were stained for CD4 and CD103. Histogram shows the CD103 expression of CD4 + Foxp3 + cells cultured with SpDCs (thick line) or LpDCs (thin line). (B) 10 5 Foxp3 − CD4 + T cells cultured with DCs and α-CD3 mAb and TGF-β as in (A) for 5 d, with the ratio of T/DC as 5/1, 10/1, 20/1 or 40/1. Data are the mean ± SD of triplicate wells. (C) Foxp3 − cells were cultured as in A with CD103 + LpDC or CD103 − LpDC. Plots are gated on CD4 + cells and Foxp3 versus α 4 β 7 stainings are shown. The numbers indicate percentage of events in each quadrant. Data are from one of at least three independent experiments.

Article Snippet: In some experiments, the following were included in the coculture conditions: (a) all-trans RA (Sigma-Aldrich) at varying concentrations; (b) RA receptor β inhibitors LE540 (Wako Chemicals USA) and LE135 (Tocris Bioscience), each at 1 μM; and (c) α-TGF-β mAb (1D11.16.8) or isotype control mAb MOPC (31C; both from American Type Culture Collection) at 10 μg/ml.

Techniques: Expressing, Cell Culture, Staining

Journal: The Journal of Experimental Medicine

Article Title: Small intestine lamina propria dendritic cells promote de novo generation of Foxp3 T reg cells via retinoic acid

doi: 10.1084/jem.20070602

Figure Lengend Snippet: Maintenance of Foxp3 contributes to higher frequency of Foxp3 + CD4 + T cells in the presence of LpDC. (A) eGFP − CD4 + T cells were cocultured with purified DC at a 5:1 ratio with α-CD3 and TGF-β. At indicated time points, cells were harvested and stained for CD4, Class II, and 7-AAD. Foxp3 expression was determined based on eGFP fluorescence. The percentage of viable cells is depicted by open bars for LpDC and filled bars for SpDC. Data are from one of two independent experiments. (B) CFSE-labeled naive CD4 + T cells were cocultured with purified DCs as described in A. On day 5, cells were harvested and stained for CD4 and intracellular Foxp3. Illustrated are dot plots of Foxp3 versus CFSE. The percentage of Foxp3 + cells and the MFI was defined as the bordered population. (C) In cultures containing LpDC (○) or SpDC (•) the proportion of Foxp3 + cells among CD4 T cells is plotted as a function of the number of cell divisions. Data are representative of three independent experiments.

Article Snippet: In some experiments, the following were included in the coculture conditions: (a) all-trans RA (Sigma-Aldrich) at varying concentrations; (b) RA receptor β inhibitors LE540 (Wako Chemicals USA) and LE135 (Tocris Bioscience), each at 1 μM; and (c) α-TGF-β mAb (1D11.16.8) or isotype control mAb MOPC (31C; both from American Type Culture Collection) at 10 μg/ml.

Techniques: Purification, Staining, Expressing, Fluorescence, Labeling

Journal: The Journal of Experimental Medicine

Article Title: Small intestine lamina propria dendritic cells promote de novo generation of Foxp3 T reg cells via retinoic acid

doi: 10.1084/jem.20070602

Figure Lengend Snippet: RA production by LpDC is requisite for optimal Foxp3 + T reg cell conversion. Foxp3 − CD4 T cells were cocultured with DC at a 10:1 ratio as described in Cells were stained for α 4 β 7 and assessed for eGFP fluorescence (Foxp3) by flow cytometry. Results are representative of three independent experiments. (B) The dose responsiveness of all-trans RA on eGFP (Foxp3) expression by CD4 T cells cocultured with SpDC was determined by flow cytometry. Error bars represent the SDs of the means of three individual samples from one experiment. Statistical significance was determined using the Student's t test. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Results are representative of three independent experiments. (C) Foxp3 − CD4 + T cells were cocultured with CD103 + or CD103 − DC at a 10:1 ratio as described in . Cells were stained for α 4 β 7 and the effect of 100 nM RA, or the RA receptor inhibitors LE540 and LE135 on (eGFP) Foxp3 expression was determined by flow cytometry. Results are representative of three independent experiments. (D) Foxp3 − CD4 T cells were cocultured with CD103 + DC in the presence of α-CD3 or α-CD3 in combination with TGF-β neutralizing antibody (isotype IgG1) or LE540 and LE135. eGFP (Foxp3) expression was determined by flow cytometry. NS. Error bars represent the SD from the means of three independent experiments. ***, P < 0.001.

Article Snippet: In some experiments, the following were included in the coculture conditions: (a) all-trans RA (Sigma-Aldrich) at varying concentrations; (b) RA receptor β inhibitors LE540 (Wako Chemicals USA) and LE135 (Tocris Bioscience), each at 1 μM; and (c) α-TGF-β mAb (1D11.16.8) or isotype control mAb MOPC (31C; both from American Type Culture Collection) at 10 μg/ml.

Techniques: Staining, Fluorescence, Flow Cytometry, Expressing

Journal: Journal of Veterinary Science

Article Title: In vitro effects of meloxicam on the number, Foxp3 expression, production of selected cytokines, and apoptosis of bovine CD25+CD4+ and CD25-CD4+ cells

doi: 10.4142/jvs.2013.14.2.125

Figure Lengend Snippet: The effect of MEL on IL-10 (A), TGF-β (B), and IFN-γ (C) production. The results are expressed as a percentage of CD25 high CD4+, CD25 low CD4+, and CD25-CD4+ cells expressing IL-10, TGF-β, or IFN-γ. Typical cytograms (D) illustrating IFN-γ expression in particular lymphocyte subpopulations are shown. Results are presented as the mean ± SE (n = 18) of three independent experiments (each one using six animals). * p < 0.01, MEL-treated cells versus the control cells.

Article Snippet: After extracellular staining (as described above) and fixation (200 μL 2% paraformaldehyde in Dulbecco's PBS per sample for 15 min on ice; both reagents were purchased from Sigma-Aldrich, USA), the cells were permeabilized with 2 mL SAP buffer [0.1% (w/v) saponin and 0.05% (w/v) NaN 3 in Hanks' balanced salt solution (HBSS; all from Sigma-Aldrich, USA)] and stained with APC (allophycocyanin) -conjugated mouse anti-TGF-β mAb (1 : 20, 1D11, IgG1, this antibody reacts with human, mouse, and bovine TGF-β1 and TGF-β2; R&D Systems, USA) for 45 min at RT in the dark.

Techniques: Expressing, Control

Journal:

Article Title: Advanced glycation end products cause epithelial-myofibroblast transdifferentiation via the receptor for advanced glycation end products (RAGE)

doi:

Figure Lengend Snippet: Exposure to AGEs causes a dose-dependent change in phenotype and antigen expression. After 6 days under experimental conditions, cultured cells were stained for α-SMA. (a) NRK 52E cells in media alone. (b) AGE-BSA (20 μM). Insert shows positive α-SMA staining in longitudinal microfilaments. (c) AGE-BSA (40 μm). (d) Unglycated BSA. (e) AGE-RNAse (100 μM). (f) AGE-BSA (40 μM) and Ab’s to the RAGE receptor, (g) TGF-β, and (h) control Ab’s. Original magnification, ×200; except insert in b, ×400.

Article Snippet: Ab’s to TGF-β (MAB 1835; R&D Systems Inc.) and E-cadherin ( {"type":"entrez-nucleotide","attrs":{"text":"C37020","term_id":"2373161","term_text":"C37020"}} C37020 ; Transduction Laboratories, San Diego, California, USA) were used at 5 μg/ml.

Techniques: Expressing, Cell Culture, Staining

Journal:

Article Title: Advanced glycation end products cause epithelial-myofibroblast transdifferentiation via the receptor for advanced glycation end products (RAGE)

doi:

Figure Lengend Snippet: Time- and dose-dependent α-SMA expression by NRK 52E cells. (a) Cells cultured in the presence of increasing concentrations of AGE-BSA or unglycated BSA, in the presence or absence of Ab’s to RAGE, TGF-β, or control Ab’s, were counted in three or more separate experiments. Represented are α-SMA +ve cells counted in ten or more high-power fields (×200) expressed as a percentage of total cell number (mean ± SEM). **P < 0.01, ***P < 0.001 compared with untreated cells. The observed increase in α-SMA staining with AGE-BSA (40 μM) was prevented by Ab’s to either RAGE (R) or to TGF-β (T), but not by control Ab’s (C). ##P < 0.01, ###P < 0.001 compared with AGE-BSA (40 μM). (b) Effects of exposure of NRK cells to AGE-BSA (40 μM) over time. Cells were examined at 2, 4, and 6 days in three or more separate experiments; **P < 0.01, ***P < 0.001 compared with 0 days. (c) Effects of exposure of NRK cells to CML-modified BSA. Variously, CML-modified BSA was prepared by incubating BSA with sodium cyanoborohydride, in the presence or absence of glyoxylic acid. Incubation of NRK cells in the presence of CML-BSA (200 μM) caused a modification-dependent increase in α-SMA staining. Shown are results of three or more independent experiments; degree of lysine modification versus SMA positive cells as percentage of total cell number (mean ± SEM). *P < 0.05, **P < 0.01.

Article Snippet: Ab’s to TGF-β (MAB 1835; R&D Systems Inc.) and E-cadherin ( {"type":"entrez-nucleotide","attrs":{"text":"C37020","term_id":"2373161","term_text":"C37020"}} C37020 ; Transduction Laboratories, San Diego, California, USA) were used at 5 μg/ml.

Techniques: Expressing, Cell Culture, Staining, Modification, Incubation

Journal:

Article Title: Advanced glycation end products cause epithelial-myofibroblast transdifferentiation via the receptor for advanced glycation end products (RAGE)

doi:

Figure Lengend Snippet: E-cadherin expression is lost in cells cultured with AGE-BSA. After 6 days under experimental conditions, cells were stained for E-cadherin. (a) NRK 52E cells grown in media alone. (b) Culture with AGE-BSA (20 μM), (c) AGE-BSA (40 μM). (d) Cells cultured with AGE-BSA (40 μM) and neutralizing Ab to RAGE. Original magnification, ×200. (e) E-cadherin quantitation. Cells expressing E-cadherin were counted in six or more high-power fields (×400) and expressed as a percentage of total cell number (mean ± SEM). Coincubation in the presence of Ab’s to RAGE (R) or TGF-β (T) abrogated the expected E-cadherin loss, whereas control Ab’s (C) did not. **P < 0.01 compared with untreated cells; ##P < 0.01 compared with AGE-BSA (40 μM) by ANOVA.

Article Snippet: Ab’s to TGF-β (MAB 1835; R&D Systems Inc.) and E-cadherin ( {"type":"entrez-nucleotide","attrs":{"text":"C37020","term_id":"2373161","term_text":"C37020"}} C37020 ; Transduction Laboratories, San Diego, California, USA) were used at 5 μg/ml.

Techniques: Expressing, Cell Culture, Staining, Quantitation Assay

Journal:

Article Title: Advanced glycation end products cause epithelial-myofibroblast transdifferentiation via the receptor for advanced glycation end products (RAGE)

doi:

Figure Lengend Snippet: Effect of AGE-BSA exposure on total TGF-β production. Total TGF-β levels were measured by ELISA (mean ± SEM). AGE-BSA, but not BSA, exposure caused a dose-dependent increase in TGF-β protein levels at 3 days that was abrogated by the presence of either Ab’s to RAGE (R) or TGF-β (T), but not by control Ab’s (C). *P < 0.05, **P < 0.01 vs. AGE-BSA (0 μM); #P < 0.05, ##P < 0.01 vs. AGE-BSA (40 μM) by ANOVA.

Article Snippet: Ab’s to TGF-β (MAB 1835; R&D Systems Inc.) and E-cadherin ( {"type":"entrez-nucleotide","attrs":{"text":"C37020","term_id":"2373161","term_text":"C37020"}} C37020 ; Transduction Laboratories, San Diego, California, USA) were used at 5 μg/ml.

Techniques: Enzyme-linked Immunosorbent Assay

Journal:

Article Title: Advanced glycation end products cause epithelial-myofibroblast transdifferentiation via the receptor for advanced glycation end products (RAGE)

doi:

Figure Lengend Snippet: Quantitation of in vivo α-SMA and TGF-β staining. Kidney sections from SD rats, control, 32-week diabetic, and 32-week diabetic + ALT 711 (n = 5 each) were examined. (a) The number of α-SMA +ve tubules counted in 20 or more high-power fields (×200; ≥ 600 tubules) is expressed as a percentage of total tubule number. ***P < 0.001 compared with control, #P < 0.05 compared with diabetic, using unpaired t test. (b) In addition, the number of individual tubular cells that stained for α-SMA in affected tubules were counted and are expressed as a percentage of total tubular cell number, ##P < 0.01 compared with diabetic. (c) TGF-β immunostaining. Diabetes was associated with a significant increase in TGF-β immunostaining that was ameliorated, but not normalized by ALT 711 treatment. The tubular TGF-β staining was quantitated by a computer-based imaging system. Results are expressed as a proportion of total area; n = 5 animals per group.

Article Snippet: Ab’s to TGF-β (MAB 1835; R&D Systems Inc.) and E-cadherin ( {"type":"entrez-nucleotide","attrs":{"text":"C37020","term_id":"2373161","term_text":"C37020"}} C37020 ; Transduction Laboratories, San Diego, California, USA) were used at 5 μg/ml.

Techniques: Quantitation Assay, In Vivo, Staining, Immunostaining, Imaging

Journal:

Article Title: Advanced glycation end products cause epithelial-myofibroblast transdifferentiation via the receptor for advanced glycation end products (RAGE)

doi:

Figure Lengend Snippet: TGF-β immunostaining in the SD rats. Diabetes was associated with a marked increase in TGF-β immunostaining (b) when compared with controls (a). This increase was ameliorated by treatment with ALT 711 (c). Representative pictures are shown at magnification ×400.

Article Snippet: Ab’s to TGF-β (MAB 1835; R&D Systems Inc.) and E-cadherin ( {"type":"entrez-nucleotide","attrs":{"text":"C37020","term_id":"2373161","term_text":"C37020"}} C37020 ; Transduction Laboratories, San Diego, California, USA) were used at 5 μg/ml.

Techniques: Immunostaining

Journal: Cell discovery

Article Title: Single-cell and spatial dissection of precancerous lesions underlying the initiation process of oral squamous cell carcinoma.

doi: 10.1038/s41421-023-00532-4

Figure Lengend Snippet: Fig. 7 mIHC staining showing clinical relevance and cellular crosstalk landscape for OSCC initiation. a, b mIHC staining of two groups of selected markers (Group 1: PD-L1, TGFβ1, VEGFA, CD68, and PanCK; Group 2: APOE, CD68, FOXP3, OX40, and PanCK) in the samples of de novo OLK (a) and recurrent OSCC with OLK (b). Those recurrent OSCC samples with recurrent OLK were taken from patients who had OLK-derived OSCC before. The patients were under a clinical trial focusing on anti-PD-1 antibody treatment. Scale bars: 200 μm. Statistical quantification is shown in each image. c Flowchart showing the induction of OSCC by 4NQO in C57BL/6 mice and intraperitoneal injection of anti-PD-1/anti-TGFβ/anti-PD-1+ anti-TGFβ antibody in each treatment group. d Representative intraoral lesions on the tongues of mice in each group. e Macroscopic lesions on the tongues of each group. The dotted circle indicates cauliflower-like lesions. f Statistical results for quantification of the macroscopic cauliflower-like lesions in each treated and untreated group. A two-tailed Student’s t-test for the P values. *P < 0.05. g Potential maps on malignant transformation of epithelial cells and dynamic crosstalk between epithelial cells and the TMEs during OSCC initiation.

Article Snippet: Mice received 200 mg anti-PD-1 antibody (InVivoMAb, BioXCell, anti-mouse-PD-1 antibody, BE0146, clone RMP1-14) and 300mg anti-TGFβ antibody (InVivoMAb, Selleck, anti-human/mouse-TGFβ antibody, A2113, clone 1D11.16.8) per injection intraperitoneally.

Techniques: Staining, Derivative Assay, Injection, Two Tailed Test, Transformation Assay

Journal: Infection and Immunity

Article Title: Expression and Function of Transforming Growth Factor ? in Melioidosis

doi: 10.1128/IAI.05534-11

Figure Lengend Snippet: TGF-β levels are elevated in melioidosis patients. (A) On admission, TGF-β levels in plasma of patients with culture-proven melioidosis (n = 33) were elevated compared to healthy controls (n = 30). (B) Plasma levels of TGF-β decreased in melioidosis patients 2 weeks after successful antibiotic treatment (n = 6). *, P < 0.05.

Article Snippet: In depletion experiments, mice were injected intraperitoneally with 200 μg of a monoclonal anti-TGF-β (1D11, anti-TGF-β1, -β2, and -β3; Bioceros) or an isotype-matched control IgG antibody (anti-β-galactosidase, G113; Bioceros) 2 h before and 48 h after bacterial inoculation ( 16 , 19 ).

Techniques:

Journal: Infection and Immunity

Article Title: Expression and Function of Transforming Growth Factor ? in Melioidosis

doi: 10.1128/IAI.05534-11

Figure Lengend Snippet: Increased expression of TGF-β in lung tissue of wild-type mice infected with B. pseudomallei. Elevated pulmonary TGF-β levels were observed in mice (n = 8) 72 h after intranasal infection with 7.5 × 102 CFU of B. pseudomallei compared to 24 h after infection. ***, P < 0.001.

Article Snippet: In depletion experiments, mice were injected intraperitoneally with 200 μg of a monoclonal anti-TGF-β (1D11, anti-TGF-β1, -β2, and -β3; Bioceros) or an isotype-matched control IgG antibody (anti-β-galactosidase, G113; Bioceros) 2 h before and 48 h after bacterial inoculation ( 16 , 19 ).

Techniques: Expressing, Infection

Journal: Infection and Immunity

Article Title: Expression and Function of Transforming Growth Factor ? in Melioidosis

doi: 10.1128/IAI.05534-11

Figure Lengend Snippet: Decreased expression of phosphorylated Smad2 in mice receiving anti-TGF-β treatment. TGF-β signaling was evaluated by quantifying the total levels of Smad2 and Smad3 and their phosphorylation in lung tissue by Western blot 48 h after intranasal infection with B. pseudomallei. The left 4 lanes show the expression of Smad2 (A) and Smad3 (B) in lung tissue of control mice and the right 4 lanes of mice that received anti-TGF-β treatment (2 × 200 μg TGFβ antibody). The bar graph shows the ratio of expressed phosphorylated Smad/total Smad corrected for actin loading levels. (C) A significant decrease in p-Smad2 is demonstrated in mice that were treated with anti-TGF-β antibody. *, P < 0.05.

Article Snippet: In depletion experiments, mice were injected intraperitoneally with 200 μg of a monoclonal anti-TGF-β (1D11, anti-TGF-β1, -β2, and -β3; Bioceros) or an isotype-matched control IgG antibody (anti-β-galactosidase, G113; Bioceros) 2 h before and 48 h after bacterial inoculation ( 16 , 19 ).

Techniques: Expressing, Western Blot, Infection

Journal: Infection and Immunity

Article Title: Expression and Function of Transforming Growth Factor ? in Melioidosis

doi: 10.1128/IAI.05534-11

Figure Lengend Snippet: Cytokine profile during experimental melioidosis in mice treated with control antibody or anti-TGF-β a

Article Snippet: In depletion experiments, mice were injected intraperitoneally with 200 μg of a monoclonal anti-TGF-β (1D11, anti-TGF-β1, -β2, and -β3; Bioceros) or an isotype-matched control IgG antibody (anti-β-galactosidase, G113; Bioceros) 2 h before and 48 h after bacterial inoculation ( 16 , 19 ).

Techniques:

Journal: Infection and Immunity

Article Title: Expression and Function of Transforming Growth Factor ? in Melioidosis

doi: 10.1128/IAI.05534-11

Figure Lengend Snippet: Less organ damage in mice receiving anti-TGF-β treatment. Levels of (A) BUN, (B) creatinine, (C) AST, and (D) ALT were measured in B. pseudomallei-infected mice (n = 8/group) that were treated with anti-TGF-β antibody and their controls at 72 h as parameters for organ damage. Expression of AST and BUN were decreased, demonstrating less organ damage. *, P < 0.05. Data shown from single independent experiments.

Article Snippet: In depletion experiments, mice were injected intraperitoneally with 200 μg of a monoclonal anti-TGF-β (1D11, anti-TGF-β1, -β2, and -β3; Bioceros) or an isotype-matched control IgG antibody (anti-β-galactosidase, G113; Bioceros) 2 h before and 48 h after bacterial inoculation ( 16 , 19 ).

Techniques: Infection, Expressing

Journal: Infection and Immunity

Article Title: Expression and Function of Transforming Growth Factor ? in Melioidosis

doi: 10.1128/IAI.05534-11

Figure Lengend Snippet: Anti-TGF-β treatment results in diminished bacterial growth during experimental melioidosis. Mice (n = 8/group) were treated with anti-TGF-β antibody or control IgG antibody and inoculated with 7.5 × 102 CFU of B. pseudomallei intranasally. Bacterial numbers were quantified 72 h postinfection. Less growth was seen in the lungs (A) and spleens (B) of mice treated with anti-TGF-β antibody. *, P < 0.05. Data shown from single independent experiments.

Article Snippet: In depletion experiments, mice were injected intraperitoneally with 200 μg of a monoclonal anti-TGF-β (1D11, anti-TGF-β1, -β2, and -β3; Bioceros) or an isotype-matched control IgG antibody (anti-β-galactosidase, G113; Bioceros) 2 h before and 48 h after bacterial inoculation ( 16 , 19 ).

Techniques:

Journal: Infection and Immunity

Article Title: Expression and Function of Transforming Growth Factor ? in Melioidosis

doi: 10.1128/IAI.05534-11

Figure Lengend Snippet: No influence on survival of anti-TGF-β treatment during murine melioidosis. Mice (n = 12/group) received either 2× 200 μg control antibody (black squares) or 2× 200 μg anti-TGF-β antibody (white squares) and were infected with 7.5 × 102 CFU, a lethal dose, of B. pseudomallei intranasally, after which they were followed until death. Mortality was assessed every 4 h. No difference in survival was seen.

Article Snippet: In depletion experiments, mice were injected intraperitoneally with 200 μg of a monoclonal anti-TGF-β (1D11, anti-TGF-β1, -β2, and -β3; Bioceros) or an isotype-matched control IgG antibody (anti-β-galactosidase, G113; Bioceros) 2 h before and 48 h after bacterial inoculation ( 16 , 19 ).

Techniques: Infection