mouse anti tgf β1 ab  (R&D Systems)

Journal: Food & function

Article Title: Lactiplantibacillus plantarum 22A-3-induced TGF-β1 secretion from intestinal epithelial cells stimulated CD103 + DC and Foxp3 + Treg differentiation and amelioration of colitis in mice.

doi: 10.1039/d1fo00990g

Figure Lengend Snippet: Fig. 1 LP22A3 attenuated TNF-α and TGF-β production, and IL-8 and TGF-β mRNA expression in the Caco-2/RAW264.7 co-culture model. LP22A3 (1 × 108 cfu mL−1) was added into the apical compartment in a Caco-2/RAW264.7 co-culture model for 3 h. Subsequently, LPS was added to the basolateral compartment at a final concentration of 7 ng mL−1 and incubated for an additional 3 h. (A) TNF-α production in the basolateral side was determined by a L929 cytotoxicity assay. (B) IL-8 mRNA expression in Caco-2 cells was detected by RT-PCR. (C) TGF-β production from Caco-2 cells was measured by ELISA. (D) TGF-β mRNA expression in Caco-2 cells was detected by Realtime-PCR, where the values represent the means ± SE (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001 (vs. LPS).

Article Snippet: To assess the effect of TLR2 and TGF-β1 neutralization on LP22A3 activity, mouse anti-TGF-β1 Ab (R&D system, Minneapolis, MN) and mouse anti-TLR2 Ab (Hycult Biotech, Plymouth Meeting, PA) were used.

Techniques: Expressing, Co-Culture Assay, Concentration Assay, Incubation, Cytotoxicity Assay, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Food & function

Article Title: Lactiplantibacillus plantarum 22A-3-induced TGF-β1 secretion from intestinal epithelial cells stimulated CD103 + DC and Foxp3 + Treg differentiation and amelioration of colitis in mice.

doi: 10.1039/d1fo00990g

Figure Lengend Snippet: Fig. 2 Effect of TGF-β1 and TLR2 on LP22A3 inhibitory properties. Anti-TGF-β1 or anti-TLR2 neutralizing antibodies were applied to the basolateral compartment of the co-culture model prior to LP22A3 treatment. (A and C) TNF-α production on the basolateral side was determined by a L929 cytotoxicity assay. (B and D) IL-8 mRNA expression in Caco-2 cells was detected by Realtime-PCR. (E) TGF-β1 mRNA expression in Caco-2 cells was detected by Realtime-PCR. Values represent the means ± SE (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001 (vs. LPS).

Article Snippet: To assess the effect of TLR2 and TGF-β1 neutralization on LP22A3 activity, mouse anti-TGF-β1 Ab (R&D system, Minneapolis, MN) and mouse anti-TLR2 Ab (Hycult Biotech, Plymouth Meeting, PA) were used.

Techniques: Co-Culture Assay, Cytotoxicity Assay, Expressing

Journal: Food & function

Article Title: Lactiplantibacillus plantarum 22A-3-induced TGF-β1 secretion from intestinal epithelial cells stimulated CD103 + DC and Foxp3 + Treg differentiation and amelioration of colitis in mice.

doi: 10.1039/d1fo00990g

Figure Lengend Snippet: Fig. 3 LP22A3 induced TGF-β1 expression in mouse ileum epithelial cells through TLR2. Three independent ligated intestinal loops were pre- treated with an isotype control or anti-TGF-β1 Ab for 30 min. A vehicle or LP22A3 sample was then injected into each loop and incubated for 2 hours. Epithelial cells were isolated, extracted for RNA, and TGF- β1 mRNA expression was detected by Realtime-PCR. Values represent the means ± SE (n = 3).

Article Snippet: To assess the effect of TLR2 and TGF-β1 neutralization on LP22A3 activity, mouse anti-TGF-β1 Ab (R&D system, Minneapolis, MN) and mouse anti-TLR2 Ab (Hycult Biotech, Plymouth Meeting, PA) were used.

Techniques: Expressing, Control, Injection, Incubation, Isolation

Journal: Food & function

Article Title: Lactiplantibacillus plantarum 22A-3-induced TGF-β1 secretion from intestinal epithelial cells stimulated CD103 + DC and Foxp3 + Treg differentiation and amelioration of colitis in mice.

doi: 10.1039/d1fo00990g

Figure Lengend Snippet: Fig. 6 Administration of LP22A3 induced TGF-β1 secretion from the IECs of the small intestine. LP22A3 was orally administrated for one week and IECs of the ileum (A) and colon (B) were cultured and measured for TGF-β1 production. Serum from untreated and LP22A3-treated mice was measured for (C) TGF-β1 and (D) IL-10 level. Values represent the means ± SE (n = 3). *P < 0.05.

Article Snippet: To assess the effect of TLR2 and TGF-β1 neutralization on LP22A3 activity, mouse anti-TGF-β1 Ab (R&D system, Minneapolis, MN) and mouse anti-TLR2 Ab (Hycult Biotech, Plymouth Meeting, PA) were used.

Techniques: Cell Culture

Journal: Food & function

Article Title: Lactiplantibacillus plantarum 22A-3-induced TGF-β1 secretion from intestinal epithelial cells stimulated CD103 + DC and Foxp3 + Treg differentiation and amelioration of colitis in mice.

doi: 10.1039/d1fo00990g

Figure Lengend Snippet: Fig. 7 Administration of LP22A3 induced anti-inflammatory cytokines expression in lymphocytes of lamina propria. LP22A3 was orally admini- strated for one week. Isolated lamina propria lymphocytes from (A) ileum and (B) colon were measured for TGF-β1, IL-10 and Foxp3 mRNA expression. Values represent the means ± SE (n = 3). *P < 0.05, **P < 0.01.

Article Snippet: To assess the effect of TLR2 and TGF-β1 neutralization on LP22A3 activity, mouse anti-TGF-β1 Ab (R&D system, Minneapolis, MN) and mouse anti-TLR2 Ab (Hycult Biotech, Plymouth Meeting, PA) were used.

Techniques: Expressing, Isolation

Journal: The Journal of Immunology Author Choice

Article Title: IDO1 and TGF-β Mediate Protective Effects of IFN-α in Antigen-Induced Arthritis

doi: 10.4049/jimmunol.1502125

Figure Lengend Snippet: The protective effect of IFN-α against AIA is mediated by TGF-β signaling in the sensitization phase. (A) Schematic representation of the AIA model, as described in Materials and Methods. Briefly, AIA was induced in WT Sv129 mice and LysM Cre+/−Tgfbr2fl/fl with or without IFN-α treatment. TGF-β signaling was blocked in WT mice by daily i.p. injection of 150 μg anti–TGF-β Ab for the first 0–8 or 20–28 d of AIA. (B) The level of arthritis is expressed as severity score (mean ± SEM, n ≥ 4) from WT mice with or without IFN-α and anti–TGF-β treatment. (C) Representative histochemical slides showing inflammation of the knee joint from each group. (D) Level of arthritis expressed as severity score (mean ± SEM, n ≥ 6) from LysM Cre+/−Tgfbr2fl/fl mice with or without IFN-α treatment. Comparison of arthritis severity score between different treatment groups was done by Mann–Whitney U test (*p < 0.05).

Article Snippet: TGF-β was neutralized by daily i.p. injection for the indicated time of 150 μg anti–TGF-β Ab (clone 1D11, reactive to mouse TGF-β1/β3; EPIRUS Biopharmaceuticals Netherlands BV).

Techniques: Injection, MANN-WHITNEY

Journal: The Journal of Immunology Author Choice

Article Title: IDO1 and TGF-β Mediate Protective Effects of IFN-α in Antigen-Induced Arthritis

doi: 10.4049/jimmunol.1502125

Figure Lengend Snippet: IFN-α–induced activation of IDO1 in the sensitization phase of AIA and in vitro occurs independently of TGF-β. Female mice were immunized days 1 and 7 with mBSA with or without IFN-α treatment, with or without anti–TGF-β Abs administered i.p. days 1–8 as in Fig. 1 and as described for AIA in Materials and Methods. (A) Sera were collected at days 9 and 28 and analyzed for Trp and Kyn concentration by HPLC. Data are expressed as the ratio of serum levels of Kyn to Trp, n ≥ 7. Comparisons between groups were made by Mann–Whitney U test. (B) Splenocytes isolated day 10 from mBSA-sensitized mice were restimulated ex vivo with 50 μg/ml mBSA with or without 100 U/ml IFN-α, with or without 40 μg/ml anti–TGF-β. Data are expressed as fold change normalized to the reference gene Actb and Ido1 expression in mBSA-stimulated cultured cells from the same mice, n = 5. Paired Student t test was used to evaluate differences between treatments. *p < 0.05.

Article Snippet: TGF-β was neutralized by daily i.p. injection for the indicated time of 150 μg anti–TGF-β Ab (clone 1D11, reactive to mouse TGF-β1/β3; EPIRUS Biopharmaceuticals Netherlands BV).

Techniques: Activation Assay, In Vitro, Concentration Assay, MANN-WHITNEY, Isolation, Ex Vivo, Expressing, Cell Culture

Journal: The Journal of Immunology Author Choice

Article Title: IDO1 and TGF-β Mediate Protective Effects of IFN-α in Antigen-Induced Arthritis

doi: 10.4049/jimmunol.1502125

Figure Lengend Snippet: IFN-α induces IDO1 expression in splenic pDC in vitro and in vivo independently of TGF-β. AIA was induced in female mice with IFN-α treatment, as described in Materials and Methods. At days 4 and 28, mice were sacrificed and pDC were purified from their spleens. (A) Western blot analysis of IDO1 protein expression in pDC. (B) Real-time PCR analysis of Ido1, Tgfb1, Ifna11, and Ifnb1 transcripts in pDC isolated from spleen cells at days 4 and 28 of AIA from mice treated with IFN-α, normalized to the expression of Actb, and presented relative to results of pDC isolated from control mice not treated with IFN-α (dotted line, 1-fold). (C) Splenocytes were isolated at day 10 of AIA, stimulated with 50 μ/ml mBSA plus 250 U/ml IFN-α, with or without anti–TGF-β (40 μg/ml) for 72 h, and analyzed for IDO1-positive pDC by FACS (see Materials and Methods). The bar diagram (left) depicts the percentage of mPDCA+IDO1+ cells among the live lymphocyte population, and the FACS plots (right) show representative dot plots from one of five mice. Data in (A) are the representative of four experiments, and (B) and (C) (mean ± SD, n = 6) were analyzed by Student t test. *p < 0.05, ***p < 0.001.

Article Snippet: TGF-β was neutralized by daily i.p. injection for the indicated time of 150 μg anti–TGF-β Ab (clone 1D11, reactive to mouse TGF-β1/β3; EPIRUS Biopharmaceuticals Netherlands BV).

Techniques: Expressing, In Vitro, In Vivo, Purification, Western Blot, Real-time Polymerase Chain Reaction, Isolation