Journal: Bioactive Materials

Article Title: A tantalum-containing zirconium-based metallic glass with superior endosseous implant relevant properties

doi: 10.1016/j.bioactmat.2024.04.014

Figure Lengend Snippet: Macrophage responses to different surface chemistry. A) ELISA results of IL-1β, TNF-α and IL-10. Ta0 and Ta3-A significantly suppressed the secretion of pro-inflammatory cytokines. B) qRT-PCR results of M1 and M2 activity related markers. Ta0 and Ta3-A significantly reduced the expression of pro-inflammatory markers. C) Heatmap of representative overlapped genes between Ta3-A vs cpTi and Ta0 vs cpTi comparisons, showing an overall decrease in pro-inflammatory genes. D) Western blot result of iNos, Cd80, Cd206 and β-actin. Data are presented as mean ± SE. * p < 0.05, ** p < 0.01 and *** p < 0.001.

Article Snippet: The membranes were blotted with 5 % non-fat dry milk (Sigma) for 1 h and then incubated overnight with CD80 Rabbit mAb (1:1000, Abclonal, Cat No: A23688), Mannose Receptor/CD206 Rabbit mAb (1:1000, Abclonal, Cat No: A21014), iNOS Polyclonal antibody (1:1000, Proteintech, Cat No: 22226-1-AP), BMP4 Polyclonal antibody (1:1000, Proteintech, Cat No: 12492-1-AP), SMAD9 Polyclonal antibody (1:1000, Proteintech, Cat No: 16397-1-AP), p -SMAD1/5/8 Rabbit mAb (1:1000, Cell Signaling, Cat No: 13820T), Alkaline Phosphate (ALPL) Rabbit mAb (1:1000, Abclonal, Cat No: A0514), Rabbit Anti-beta-Actin/HRP antibody (1:1000, Bioss, Cat No: bsm-52846R), followed by incubation with a goat anti-rabbit IgG secondary antibody HRP conjugated (1:1000, Beyotime, Cat No: A0208).

Techniques: Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Activity Assay, Expressing, Western Blot

Journal: Scientific Reports

Article Title: A novel and effective method to generate human porcine-specific regulatory T cells with high expression of IL-10, TGF-β1 and IL-35

doi: 10.1038/s41598-017-04322-3

Figure Lengend Snippet: DC generation from monocytes with anti-inflammatory cytokines exhibited tolerogenic phenotype and produced anti-inflammatory cytokines. TolDC and C5-DC were harvested after maturation and characterized by flow cytometry, anti-CD14, anti-CD83, anti-CD80, anti-CD86 anti-B7-H1 and anti-B7-DC antibodies were used. ( a ) Histogram, ( b ) dot chart of MFI ratio tolDC to C5-DC. Data represent 26 independent experiments from 9 donors (CD86: n = 8). Wilcoxon signed-rank test was used to determine p values. ( c ) IL-10, IL-12p70, TGF-β1 and IFN-γ quantification in DC supernatants using BD CBA Flex Set and ELISA. These data represent 10–19 independent experiments from 9 donors. Culture medium plus maturation cocktail without cells were used as control. Turkey test was used to determine the statistical significance. ***p < 0.001. RNA was isolated from tolDC and C5-DC, and quantified by RT-PCR, ( d ) Relative RNA level of IL-35 and related genes, IL-10 , TGFB1 in tolDC, C5-DC as control. ACTB , G6PDH or CYPB mRNA were determined from the same sample and used as endogenous reference genes. These data represent 7 independent experiments from 6 donors. Wilcoxon signed-rank test was used to determine p values. Error bars: s.e.m.

Article Snippet: For immunophenotyping of tolDC and C5-DC, the following antibodies were used: anti-CD14 (clone: M5E2), anti-CD80 (clone: L307.4), anti-CD83 (clone: HB15e), anti-B7-H1 (clone: MIH1), anti-B7-DC (clone: MIH18) (all from BD Biosciences, USA) and anti-CD86 (clone: 2331, Pharmingen).

Techniques: Produced, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Isolation, Reverse Transcription Polymerase Chain Reaction

Journal: Drug Design, Development and Therapy

Article Title: Osteopontin promotes dendritic cell maturation and function in response to HBV antigens

doi: 10.2147/DDDT.S81656

Figure Lengend Snippet: Blockage of OPN suppressed DC maturation and cytokine production after hepatitis B virus (HBV) antigenic stimulations in human PBMC. Notes: After administering OPN neutralizing antibody in vitro, the histograms ( A ) and expressions ( B ) of CD80, CD86, and HLA-DR in BDCA-1 + cells after HBV antigenic stimulations in human PBMCs are presented. Overlay plots are representative and show isotype control (shadow) and the expressions of the indicated molecules on DCs in the absence (green line) or presence (red line) of neutralizing antibody in each group. Levels of IL-12 ( C ), TNF-α, IFN-γ, and IL-4 ( D ) in human PBMC supernatant were measured after administering the OPN-neutralizing antibody in vitro as well as HBcAg and HBsAg stimulations. * P <0.05. Abbreviations: OPN, osteopontin; DCs, dendritic cells; HBV, hepatitis B virus; HBcAg, hepatitis B core antigen; HBsAg, hepatitis B surface antigen; PBMC, peripheral blood mononuclear cells; IL, interleukin; TNF, tumor necrosis factor; IFN, interferon; Ab, antibody; IgG, Immunoglobulin gamma; HLA-DR, human leukocyte antigen-DR; BDCA-1, blood dendritic cell antigen 1.

Article Snippet: Expressions of surface molecules on DC were detected by flow cytometry using the following antibodies: PE-anti-mouse I-Ab/I-Ed (M5/114), PE-anti-mouse B7-1 (1G10), PE-anti-mouse B7-2 (Gl-1), FITC-anti-mouse CD11c (HL3), APC-anti-mouse TCRβ (H57-597), FITC-anti-human CD80 (L307.4), FITC-anti-human CD86 (2331 (FUN-1)), FITC-anti-human human leukocyte antigen DR (HLA-DR) (G46-6), PE-anti-human BDCA-1 (F10/21A3) and APC-anti-human CD3 (HIT3A) (all from BD Biosciences, San Diego, CA, USA).

Techniques: In Vitro

Journal: Drug Design, Development and Therapy

Article Title: Osteopontin promotes dendritic cell maturation and function in response to HBV antigens

doi: 10.2147/DDDT.S81656

Figure Lengend Snippet: OPN deficiency impaired DCs maturation and function after HBV antigenic stimulation in vitro. Notes: DCs derived from BM of the WT and OPN −/− mice were stimulated by HBV antigens. The histograms ( A ) and percentages ( B–D ) of CD80, CD86, and MHC-II molecules on the DC surface were detected by flow cytometry. Overlay plots are representative and show isotype control (shadow) and the expressions of the indicated molecules on DCs from WT mice (green line) or OPN −/− mice (red line) in each group. ( E ) Levels of IL-12 in the BMDC culture supernatants following different stimulations were measured by ELISA. * P <0.05; ** P <0.01. Abbreviations: OPN, osteopontin; DCs, dendritic cells; HBV, hepatitis B virus; BM, bone marrow; WT, wild-type; HBcAg, hepatitis B core antigen; HBsAg, surface antigen of the hepatitis B virus; MHC, major histocompatibility complex; IL, interleukin; BMDC, bone marrow-derived dendritic cell; ELISA, enzyme-linked immunosorbent assay.

Article Snippet: Expressions of surface molecules on DC were detected by flow cytometry using the following antibodies: PE-anti-mouse I-Ab/I-Ed (M5/114), PE-anti-mouse B7-1 (1G10), PE-anti-mouse B7-2 (Gl-1), FITC-anti-mouse CD11c (HL3), APC-anti-mouse TCRβ (H57-597), FITC-anti-human CD80 (L307.4), FITC-anti-human CD86 (2331 (FUN-1)), FITC-anti-human human leukocyte antigen DR (HLA-DR) (G46-6), PE-anti-human BDCA-1 (F10/21A3) and APC-anti-human CD3 (HIT3A) (all from BD Biosciences, San Diego, CA, USA).

Techniques: In Vitro, Derivative Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Drug Design, Development and Therapy

Article Title: Osteopontin promotes dendritic cell maturation and function in response to HBV antigens

doi: 10.2147/DDDT.S81656

Figure Lengend Snippet: OPN produced by DCs plays an important role in response to HBV antigenic stimulation in vivo. Notes: At 24 hours after HepG2.2.15 cells supernatant direct injection into liver from the WT mice and OPN −/− mice ( A ) liver histology was assessed. Original magnification, 400×. ( B ) Leukocyte number and ( C ) expressions of CD80, CD86, and MHC-II molecules on DCs in liver from the WT mice and OPN −/− mice were measured. ( D ) Number of TCR + cells per gram of liver tissue were detected by an automated cell counter and flow cytometry. ( E ) Number of IFN-γ-producing T-cells and IL-4-producing T-cells were detected. HepG2 supernatant injection was used as control. n=5 per group. Adoptive transfer of CD11c + cells from the WT and OPN −/− mice as well as HepG2.2.15 supernatant direct injection into mice liver were performed. Then ( F ) the number of leukocyte per gram of liver tissue was counted in the OPN −/− mice at 0 and 24 hours; n=5 per group. ( G ) IFN-γ-producing cells in T-cells in liver from the OPN −/− mice were analyzed. n=5 per group. * P <0.05; ** P <0.01; *** P <0.001. Abbreviations: OPN, osteopontin; DCs, dendritic cells; HBV, hepatitis B virus; WT, wild-type; MHC, major histocompatibility complex; IL, interleukin; IFN, interferon; TCR, T-cell receptor; h, hour.

Article Snippet: Expressions of surface molecules on DC were detected by flow cytometry using the following antibodies: PE-anti-mouse I-Ab/I-Ed (M5/114), PE-anti-mouse B7-1 (1G10), PE-anti-mouse B7-2 (Gl-1), FITC-anti-mouse CD11c (HL3), APC-anti-mouse TCRβ (H57-597), FITC-anti-human CD80 (L307.4), FITC-anti-human CD86 (2331 (FUN-1)), FITC-anti-human human leukocyte antigen DR (HLA-DR) (G46-6), PE-anti-human BDCA-1 (F10/21A3) and APC-anti-human CD3 (HIT3A) (all from BD Biosciences, San Diego, CA, USA).

Techniques: Produced, In Vivo, Injection, Flow Cytometry, Adoptive Transfer Assay

Journal: Drug Design, Development and Therapy

Article Title: Osteopontin promotes dendritic cell maturation and function in response to HBV antigens

doi: 10.2147/DDDT.S81656

Figure Lengend Snippet: OPN administration significantly promoted the maturation of DCs from CHB patients after HBcAg stimulation. Notes: The expressions of CD80 ( A ), CD86 ( B ), and HLA-DR ( C ) in BDCA-1 + cells from the peripheral blood of CHB patients following HBcAg stimulation with or without exogenous OPN administration were measured by flow cytometry. CHB patients, n=20. ** P <0.01, *** P <0.001. ( D ) The percentages of IFN-γ-producing cell in CD3 + T-cells from the peripheral blood of CHB patients following HBcAg stimulation with or without exogenous OPN administration were measured by flow cytometry. Abbreviations: OPN, osteopontin; DCs, dendritic cells; CHB, chronic hepatitis B; HBV, hepatitis B virus; HBcAg, hepatitis B core antigen; IFN, interferon; HLA-DR, human leukocyte antigen-DR; BDCA-1, Blood Dendritic Cell Antigen 1.

Article Snippet: Expressions of surface molecules on DC were detected by flow cytometry using the following antibodies: PE-anti-mouse I-Ab/I-Ed (M5/114), PE-anti-mouse B7-1 (1G10), PE-anti-mouse B7-2 (Gl-1), FITC-anti-mouse CD11c (HL3), APC-anti-mouse TCRβ (H57-597), FITC-anti-human CD80 (L307.4), FITC-anti-human CD86 (2331 (FUN-1)), FITC-anti-human human leukocyte antigen DR (HLA-DR) (G46-6), PE-anti-human BDCA-1 (F10/21A3) and APC-anti-human CD3 (HIT3A) (all from BD Biosciences, San Diego, CA, USA).

Techniques: Flow Cytometry