Journal: Microorganisms

Article Title: GroEL Secreted from Bacillus subtilis Natto Exerted a Crucial Role for Anti-Inflammatory IL-10 Induction in THP-1 Cells

doi: 10.3390/microorganisms11051281

Figure Lengend Snippet: SDS-PAGE of the affinity-purified GroEL ( A ). Reduction of IL-10-inducing activity of GroEL by the treatments with anti-TLR2 and -TLR4 antibodies ( B ). Significant difference from the control (-), * p < 0.05 (n = 3).

Article Snippet: For receptor analysis, an anti-TLR2 antibody (Sino Biological Inc., Peking, China) or anti-TLR4 antibody (Sino Biological Inc., Beijing, China) was pre-incubated with THP-1 DC prior to the addition of purified GroEL.

Techniques: SDS Page, Affinity Purification, Activity Assay, Control

Journal: Molecular medicine reports

Article Title: Expression of Toll‑like receptor 4 on mast cells in gingival tissues of human chronic periodontitis.

doi: 10.3892/mmr.2018.8648

Figure Lengend Snippet: Figure 6. TLR4‑positive mast cells. Total numbers of TLR4‑positive mast cells in the healthy control (n=20), mild chronic periodontitis group (n=23) and advanced chronic periodontitis group (n=25). Data are presented as the mean ± standard deviation. *P<0.05, vs. other groups. TLR4, Toll‑like receptor 4.

Article Snippet: The dilution of the I anti-TLR4 antibody (cat. no. TA309486, OriGene Technologies) was 1:100.

Techniques: Control, Standard Deviation

Journal: Molecular medicine reports

Article Title: Expression of Toll‑like receptor 4 on mast cells in gingival tissues of human chronic periodontitis.

doi: 10.3892/mmr.2018.8648

Figure Lengend Snippet: Figure 3. Expression of TLR4 in human gingival tissues. Immunohistochemical staining was performed to determine expression of TLR4 (magnification, x400). (A) Healthy control; (B) mild chronic periodontitis group; (C) advanced chronic periodontitis group. TLR4, Toll‑like receptor 4.

Article Snippet: The dilution of the I anti-TLR4 antibody (cat. no. TA309486, OriGene Technologies) was 1:100.

Techniques: Expressing, Immunohistochemical staining, Staining, Control

Journal: Molecular medicine reports

Article Title: Expression of Toll‑like receptor 4 on mast cells in gingival tissues of human chronic periodontitis.

doi: 10.3892/mmr.2018.8648

Figure Lengend Snippet: Figure 5. Expression of TLR4 on mast cells in human gingival tissues. Immunofluorescent staining was performed (magnification, x400). (A) Healthy control; (B) mild chronic periodontitis group; (C) advanced chronic periodontitis group. TLR4, Toll‑like receptor 4.

Article Snippet: The dilution of the I anti-TLR4 antibody (cat. no. TA309486, OriGene Technologies) was 1:100.

Techniques: Expressing, Staining, Control

Journal: Molecular medicine reports

Article Title: Expression of Toll‑like receptor 4 on mast cells in gingival tissues of human chronic periodontitis.

doi: 10.3892/mmr.2018.8648

Figure Lengend Snippet: Figure 4. Expression of TLR. Positive cell rates of TLR4 in the (A) healthy control (n=20), (B) mild chronic periodontitis group (n=23) and (C) advanced chronic periodontitis group (n=25). Data are presented as the mean ± standard deviation. *P<0.05, vs. other groups. TLR4, Toll‑like receptor 4.

Article Snippet: The dilution of the I anti-TLR4 antibody (cat. no. TA309486, OriGene Technologies) was 1:100.

Techniques: Expressing, Control, Standard Deviation

Journal: Experimental Animals

Article Title: Transient receptor potential vanilloid 1 interacts with Toll-like receptor 4 (TLR4)/cluster of differentiation 14 (CD14) signaling pathway in lipopolysaccharide-mediated inflammation in macrophages

doi: 10.1538/expanim.23-0148

Figure Lengend Snippet: Upon lipopolysaccharide (LPS) challenge in macrophages, the interaction of transient receptor potential vanilloid 1 (TRPV1) with Toll-like receptor 4 (TLR4) complex was increased with the formation of TLR4–myeloid differentiation protein 88 (MyD88)–IL-1 receptor-associated kinase 1 (IRAK1). (A) Whole-cell lysates were obtained from bone marrow-derived wild-type (WT) macrophages in the presence or absence of LPS (1 µ g/ml) for 5 min. Total cell lysates were immunoprecipitated with an anti-TRPV1 antibody. The blot was then immunoblotted with an anti-TLR4 antibody and an anti-CD14 antibody. (B) Quantitative results of panel A. (C) Whole-cell lysates prepared from bone marrow-derived WT and Trpv1 −/− macrophages with or without LPS treatment (1 µ g /ml) for 5 min were immunoprecipitated with anti-MyD88 antibody. The blot was then immunoblotted with an anti-TLR4 antibody and an anti-IRAK1 antibody. (D) Quantitative results of panel A. Data are mean ± SEM from four independent experiments. * P <0.05 versus WT control group, # P <0.05 versus WT LPS-treated group.

Article Snippet: Rabbit anti-TLR4 antibody was from Assay Designs (Ann Arbor, MI, USA).

Techniques: Derivative Assay, Immunoprecipitation, Control

Journal: Experimental Animals

Article Title: Transient receptor potential vanilloid 1 interacts with Toll-like receptor 4 (TLR4)/cluster of differentiation 14 (CD14) signaling pathway in lipopolysaccharide-mediated inflammation in macrophages

doi: 10.1538/expanim.23-0148

Figure Lengend Snippet: The proposed mechanism for the involvement of transient receptor potential vanilloid 1 (TRPV1) receptor in lipopolysaccharide (LPS)-induced induction of pro-inflammatory mediators in macrophages. As shown, genetic ablation of the TRPV1 receptor causes a reduction in the formation of Toll-like receptor 4 (TLR4)/cluster of differentiation 14 (CD14)/myeloid differentiation protein 88 (MyD88)/IL-1 receptor-associated kinase 1 (IRAK1) complex and the phosphorylation of inhibitor of nuclear factor kappa-Bα (IκBα) and mitogen-activated protein kinases (MAPKs) (p38, ERK1/2, and JNK1/2), and thus decreases activation of nuclear factor kappa-B (NF-κB) and activator protein 1 (AP-1), leading to downregulation of pro-inflammatory mediators including tumor necrosis factor-α (TNF-α), IL-1β, and inducible nitric oxide synthase (iNOS).

Article Snippet: Rabbit anti-TLR4 antibody was from Assay Designs (Ann Arbor, MI, USA).

Techniques: Activation Assay

Journal: Arthritis Research & Therapy

Article Title: Between adaptive and innate immunity: TLR4-mediated perforin production by CD28 null T-helper cells in ankylosing spondylitis

doi: 10.1186/ar1840

Figure Lengend Snippet: Messenger RNA expression of TLR2 and TLR4 in CD3 + CD4 + CD28 null and CD3 + CD4 + CD28 + T cells. (a) Fluorescence-activated cell sorting analysis shows the purity of CD3 + CD4 + CD28 null and CD3 + CD4 + CD28 + T cells. (b) mRNA expression of TLR2, TLR4 and β2-microglobulin (β2m, housekeeping gene) in CD3 + CD4 + CD28 null T cells (CD28 - ) and in CD3 + CD4 + CD28 + T cells (CD28 + ). Peripheral blood mononuclear cells were used as positive control (pos co), and a negative control (neg co) was performed in the absence of cDNA. A representative example out of three independent experiments is given.

Article Snippet: To test inhibitory effects of blocking anti-CD14 or anti-TLR4 antibodies, cells were resuspended in RPMI 1640 with 5% autologous serum and were incubated with 10 μg/ml blocking anti-CD14 antibody (R&D Systems), 10 μg/ml anti-TLR4 antibody (Torrey Pines Biolabs, TX, USA) and 10 μg/ml isotype control antibody (Becton Dickinson) for 1 hour before adding LPS at a concentration of 10 μg/ml.

Techniques: RNA Expression, Fluorescence, FACS, Expressing, Positive Control, Negative Control

Journal: Arthritis Research & Therapy

Article Title: Between adaptive and innate immunity: TLR4-mediated perforin production by CD28 null T-helper cells in ankylosing spondylitis

doi: 10.1186/ar1840

Figure Lengend Snippet: Surface expression of CD14, TLR4 and TLR2 on CD4 + CD28 + and CD28 null cells in ankylosing spondylitis, psoriatic arthritis and rheumatoid arthritis. (a) Representative dot plots and histograms show TLR4 expression (filled red curve, black line represents isotype control) on CD4 + CD28 + and CD4 + CD28 null T cells. Gates were set on lymphocytes (forward scatter and sideward scatter) as well as on CD28 + and CD28 null cells expressing high levels of CD4. (b) Box plots summarize the expression of CD14, TLR4 and TLR2 on CD4 + CD28 + and CD28 null T cells in patients as indicated. The Wilcoxon test was used to determine the statistical differences between the groups. *** P < 0.001. SSC, side scatter; FSC, forward scatter; AS, ankylosing spondylitis; PsA, psoriatic arthritis; RA, rheumatoid arthritis.

Article Snippet: To test inhibitory effects of blocking anti-CD14 or anti-TLR4 antibodies, cells were resuspended in RPMI 1640 with 5% autologous serum and were incubated with 10 μg/ml blocking anti-CD14 antibody (R&D Systems), 10 μg/ml anti-TLR4 antibody (Torrey Pines Biolabs, TX, USA) and 10 μg/ml isotype control antibody (Becton Dickinson) for 1 hour before adding LPS at a concentration of 10 μg/ml.

Techniques: Expressing, Control

Journal: Arthritis Research & Therapy

Article Title: Between adaptive and innate immunity: TLR4-mediated perforin production by CD28 null T-helper cells in ankylosing spondylitis

doi: 10.1186/ar1840

Figure Lengend Snippet: CD14 and TLR4-mediated effects. (a) ELISA assays were performed to analyse levels of soluble CD14 (sCD14) in sera from patients with ankylosing spondylitis (AS) ( n = 50) and healthy controls (CO) ( n = 23). The Mann-Whitney test was used to determine the statistical differences between the group of patients and the control group. ** P < 0.01. A blocking antibody (Ab) directed against (b) CD14 and (c) TLR4 or an isotype control were added to peripheral blood mononuclear cells from patients with AS maintained in 5% autologous serum. After 1 hour, lipopolysaccharide (LPS) stimulation at a concentration of 10 μg/ml for 16 hours was started. Box blots show percentages of perforin-producing CD4 + CD28 null T cells from seven independent experiments. Differences were tested for significance using the Wilcoxon test. ** P < 0.01.

Article Snippet: To test inhibitory effects of blocking anti-CD14 or anti-TLR4 antibodies, cells were resuspended in RPMI 1640 with 5% autologous serum and were incubated with 10 μg/ml blocking anti-CD14 antibody (R&D Systems), 10 μg/ml anti-TLR4 antibody (Torrey Pines Biolabs, TX, USA) and 10 μg/ml isotype control antibody (Becton Dickinson) for 1 hour before adding LPS at a concentration of 10 μg/ml.

Techniques: Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Control, Blocking Assay, Concentration Assay

Journal: Arthritis Research & Therapy

Article Title: Between adaptive and innate immunity: TLR4-mediated perforin production by CD28 null T-helper cells in ankylosing spondylitis

doi: 10.1186/ar1840

Figure Lengend Snippet: Effects of TNF-α on expression of pattern recognition receptors in vitro and in vivo . (a) Peripheral blood mononuclear cells were stimulated with 20 ng/ml tumour necrosis factor-α (TNF-α) for 24 hours, and CD4 + CD28 null T cells were analysed for expression of CD14, TLR4 and TLR2. Box plots summarize data from seven independent experiments. Medians were compared using the Wilcoxon test. *** P < 0.001, ** P < 0.01. (b) CD4 + CD28 null T cells in patients with active ankylosing spondylitis treated with infliximab at a dosage of 3 mg/kg body weight were tested for the expression of pattern recognition receptors (PRRs) before and 3 weeks after injection ( n = 8). The expression of CD14, TLR4 and TLR2 was detected by flow cytometry. The Wilcoxon test was used to determine differences in expression of PRRs before (pre) and under successful TNF-α blocking treatment (post). ** P < 0.01, * P < 0.05.

Article Snippet: To test inhibitory effects of blocking anti-CD14 or anti-TLR4 antibodies, cells were resuspended in RPMI 1640 with 5% autologous serum and were incubated with 10 μg/ml blocking anti-CD14 antibody (R&D Systems), 10 μg/ml anti-TLR4 antibody (Torrey Pines Biolabs, TX, USA) and 10 μg/ml isotype control antibody (Becton Dickinson) for 1 hour before adding LPS at a concentration of 10 μg/ml.

Techniques: Expressing, In Vitro, In Vivo, Injection, Flow Cytometry, Blocking Assay