Journal: PLoS ONE

Article Title: Novel PDE4 Inhibitors Derived from Chinese Medicine Forsythia

doi: 10.1371/journal.pone.0115937

Figure Lengend Snippet: A. 5×10 5 RAW264.7 cells were seeded in 96 wells for 18 h. Cells were primed with compounds at different concentration for 3 h before treated with LPS (1 ng/ml) for additional 8 h. TNF cytokine releases were monitored by ELISA. B. PBMC (0.2 ml at 1×10 5 /ml) were primed with compounds at different concentration for 3 h before treated with LPS (1 ng/ml) for additional 8 h. TNF cytokine releases were monitored by ELISA. % of TNF secretion were calculated and graphed. C. Summary of compound IC 50 . The data represent n = 3–6 experiments.

Article Snippet: IL1β, TNFα, IL6 mouse ELISA kit, human TNFα were from R&D systems.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Mucosal Immunology

Article Title: TNF hampers intestinal tissue repair in colitis by restricting IL-22 bioavailability

doi: 10.1038/s41385-022-00506-x

Figure Lengend Snippet: A Humanized TNF colitis model. Rag1 −/− or hTNF‐KI x Rag1 −/− mice were reconstituted with naive T cells from either WT or hTNF‐KI mice. Mice were treated once they had lost > 5 % of their initial weight twice per week for two or three weeks with various anti-TNF agents or respective controls (10 mg/kg; i.p.). B Weight of hTNF‐KIxRag1 −/− mice reconstituted with naive T cells from hTNF‐KI mice and treated with infliximab ( n = 6) or Fc control ( n = 5) (10 mg/kg) twice per week. Data are representative of two independent experiments. C Colitis inflammation scores of hTNF‐KIxRag1 −/− mice reconstituted with naive T cells from hTNF‐KI mice upon 2 weeks treatment with infliximab ( n = 4) or isotype control ( n = 8). Data are representative of two independent experiments. D Colitis inflammation scores of hTNF‐KIxRag1 −/− mice reconstituted with naive T cells from hTNF‐KI mice upon 3 weeks treatment with infliximab ( n = 6) or isotype control ( n = 5). Frequencies of CD11b + (CD45 + CD11b + ) E , Th1 (CD45 + CD4 + TCRβ + IFNγ + ) F , Th17 (CD45 + CD4 + TCRβ + IL-17A + ) G and Th1/17 (CD45 + CD4 + TCRβ + IL-17A + IFNγ + ) H cells after 3 weeks of infliximab ( n = 6) or Fc control treatment ( n = 5). All data in D – H are representative of two independent experiments. Data represent mean values ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, as calculated by Student’s t‐test, ns not significant.

Article Snippet: Once mice had lost >5% of initial weight, they were treated with anti-human TNF antibody (Infliximab), anti-mouse IL-22 (R&D Systems; AF582) or human Fc-IgG1 isotype control (BioXCell; Cat # BE0096) in PBS.

Techniques: Control

Journal: Mucosal Immunology

Article Title: TNF hampers intestinal tissue repair in colitis by restricting IL-22 bioavailability

doi: 10.1038/s41385-022-00506-x

Figure Lengend Snippet: Naive hTNF-KI T cells were transferred to Rag1 −/− recipients, anti-TNF (infliximab; 10 mg/kg; i.p. twice per week) was administered once mice had lost 5% of their initial weight. A Weight changes 2 weeks after treatment of colitic mice with either Fc control or infliximab (both 10 mg/kg; i.p, twice per week). Representative images of Hematoxylin/Eosin stained tissue sections B and inflammation score C of the colon in mice treated with either Fc control of infliximab for 2 weeks. Scale bar is equal to 100 µm. A – C Data from three independent experiments are shown. D Weight changes 3 weeks after treatment of colitic mice with either Fc control or infliximab (both 10 mg/kg; i.p, twice per week). Representative images of Hematoxylin/Eosin stained tissue sections E and inflammation score F of the colon in mice treated with either Fc control of infliximab for 3 weeks. Scale bar is equal to 100 µm. D – F Data from two independent are shown. Naive WT T cells were transferred to hTNFK-KI×Rag1 −/− recipients, anti-TNF (infliximab; 10 mg/kg; i.p. twice per week) was administered once mice had lost 5% of their initial weight. G Weight changes 2 weeks after treatment of colitic mice with either Fc control or infliximab (both 10 mg/kg; i.p, twice per week). Representative images of Hematoxylin/Eosin stained tissue sections H and inflammation score ( I ) of the colon in mice treated with either Fc control or infliximab for 2 weeks. Scale bar is equal to 100 µm. Data are representative of two independent experiments. Data represent mean values ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, as calculated by Student’s t-test; ns not significant.

Article Snippet: Once mice had lost >5% of initial weight, they were treated with anti-human TNF antibody (Infliximab), anti-mouse IL-22 (R&D Systems; AF582) or human Fc-IgG1 isotype control (BioXCell; Cat # BE0096) in PBS.

Techniques: Control, Staining

Journal: Mucosal Immunology

Article Title: TNF hampers intestinal tissue repair in colitis by restricting IL-22 bioavailability

doi: 10.1038/s41385-022-00506-x

Figure Lengend Snippet: Naive hTNF-KI T cells were transferred to Rag1 −/− recipients, anti-TNF (infliximab; 10 mg/kg; i.p. twice per week) was administered once mice lost 5% of their initial weight. A Representative images of colon tissue sections stained immunohistochemically for proliferation (Ki67, brown) and for CD3 expression (red) depict increased proliferation of colonic epithelial cells after 2 weeks of anti-TNF therapy. Scale bar is equal to 100 µm in x100 magnification and 25 µm in x400 magnification. B Increased frequencies of Ki67-positive epithelial cells and mean numbers of epithelial cells per crypt after 2 weeks of anti-TNF therapy as described in A . C Representative images of colon tissue sections stained immunohistochemically for proliferation (Ki67, red) and for CD3 expression (brown) show proliferation of colonic epithelial cells after 3 weeks of anti-TNF therapy. Scale bar is equal to 100 µm in x100 magnification and 25 µm in x400 magnification. D Frequencies of Ki67-positive epithelial cells and mean numbers of epithelial cells per crypt after 3 weeks of anti-TNF therapy as described in C . All data are representative of two independent experiments. Data represent mean values ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, as calculated by Student’s t-test; ns not significant.

Article Snippet: Once mice had lost >5% of initial weight, they were treated with anti-human TNF antibody (Infliximab), anti-mouse IL-22 (R&D Systems; AF582) or human Fc-IgG1 isotype control (BioXCell; Cat # BE0096) in PBS.

Techniques: Staining, Expressing

Journal: Mucosal Immunology

Article Title: TNF hampers intestinal tissue repair in colitis by restricting IL-22 bioavailability

doi: 10.1038/s41385-022-00506-x

Figure Lengend Snippet: A Numbers of goblet cells in the colon as revealed by PAS staining after 2 weeks of anti-TNF therapy. Data are representative of two independent experiments. B Numbers of apoptotic EC (Casp3 + ) per 100 EC in the colon after 2 weeks of anti-TNF therapy. Data collected using two independent experiments are shown. C , D . Representative images of colon sections stained immunohistochemically for pSTAT3 (brown) show increased pSTAT3 + epithelial cells upon anti-TNF therapy. B – D Data collected using two independent experiments are shown. Naive hTNF-KI T cells were transferred to Rag1 −/− recipients, anti-TNF (infliximab; 10 mg/kg; i.p. twice per week) was administered once mice lost 5% of their initial weight. Scale bar is equal to 100 µm. E Expression of STAT3 dependent genes ( Reg3β , Reg3γ , survivin , smoothened ) in the colon after 2 weeks of anti-TNF therapy. Data are representative of two independent experiments. F Gene sets upregulated in colonic EC (sorted live EpCAM1+ cells) 24 h after infliximab treatment, when compared to Fc-control treated group. G Gene sets downregulated in colonic EC (sorted live EpCAM1 + cells) 24 h after infliximab treatment, when compared to Fc-control treated group. F , G Data are from one experiment using epithelial cells from individual mice. Data represent mean values ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, as calculated by Student’s t-test; ns not significant.

Article Snippet: Once mice had lost >5% of initial weight, they were treated with anti-human TNF antibody (Infliximab), anti-mouse IL-22 (R&D Systems; AF582) or human Fc-IgG1 isotype control (BioXCell; Cat # BE0096) in PBS.

Techniques: Staining, Expressing, Control

Journal: Mucosal Immunology

Article Title: TNF hampers intestinal tissue repair in colitis by restricting IL-22 bioavailability

doi: 10.1038/s41385-022-00506-x

Figure Lengend Snippet: A Levels of IL-22 in ex vivo colonic explants from mice treated with Fc control or infliximab for 2 weeks. B IL-22 mRNA levels in the colons of mice treated with Fc control or infliximab for 2 weeks. C IL-22BP mRNA levels in the colons of mice treated with Fc control or infliximab for 2 weeks. D Serum IL-22BP concentration in sera, E Ratio of IL-22 and IL-22BP mRNA levels in the colons of mice treated with Fc control or infliximab for 2 weeks. Naive hTNF-KI T cells were transferred to Rag1 −/− recipients, anti-TNF (infliximab; 10 mg/kg; i.p. twice per week) was administered once mice lost 5% of their initial weight. F Representative images of colon sections stained with Hematoxylin/Eosin and immunohistochemically stained for Ki67 (brown) and CD3 (red), G weight changes, H corresponding inflammation scores, I percentage of proliferating colonic epithelial cells after treatment of colitic mice with either Fc control, infliximab, anti-IL-22 or infliximab/anti-IL-22 (Fc control and infliximab - 10 mg/kg; anti-IL-22 - 10 mcg/mouse; i.p, twice per week) for 2 weeks. Naive hTNF-KI T cells were transferred to Rag1 −/− recipients, antibodies were administered once mice had lost 5% of their initial weight. Scale bar is equal to 100 µm in x100 magnification and 25 µm in x400 magnification. J Colonic inflammation score at various time points after start of anti-mTNF treatment. Percentage of proliferating colonic epithelial cells K , IL-22BP mRNA levels L in the colons of mice treated with Fc control or anti-mTNF for 2 weeks. All data are representative of two independent experiments. Data represent mean values ± SEM. * p < 0.05, ** p < 0.01,*** p < 0.001, as calculated by Student’s t-test; ns not significant.

Article Snippet: Once mice had lost >5% of initial weight, they were treated with anti-human TNF antibody (Infliximab), anti-mouse IL-22 (R&D Systems; AF582) or human Fc-IgG1 isotype control (BioXCell; Cat # BE0096) in PBS.

Techniques: Ex Vivo, Control, Concentration Assay, Staining

Journal: Mucosal Immunology

Article Title: TNF hampers intestinal tissue repair in colitis by restricting IL-22 bioavailability

doi: 10.1038/s41385-022-00506-x

Figure Lengend Snippet: A TNF blockade increases levels of bioactive colonic IL-22 that can be inhibited by recombinant IL-22BP. Supernatants derived from colonic explants of Rag1 −/− mice which received naive hTNF-KI T cells and treated for 2 weeks with anti-TNF, were added to human colonic Colo205 cell line with or without rmIL-22BP (1,25 mg/ml). Production of human IL-10 as an indicator of bioactive IL-22 has been determined 48 h later. B Expression of IL-22BP mRNA in sorted CD45 + CD11c + MHCII + CD103 + and CD45 + TCRβ + cells from the colon of mice treated with Fc control or infliximab for 2 weeks. Naive hTNF-KI T cells were transferred to Rag1 −/− recipients, anti-TNF (infliximab; 10 mg/kg; i.p. twice per week) was administered once mice had lost 5% of their initial weight. C Expression of IL-22BP in colon and spleen from naive WT and TNF −/− mice. D Expression of IL-22BP in spleen from naive WT, T-TNF −/− , B-TNF −/− , and M-TNF −/− mice. E Expression of IL-22BP in sorted CD4 + CD11c + MHCII + or CD8 + CD11c + MHCII + cells isolated from spleen of naïve WT and TNF −/− mice. CD4 + CD11c + MHCII + cells from TNF −/− mice were incubated with rmTNF (100 ng/ml; 4 h). F Levels of human IL-22BP in human monocyte-derived DCs (moDCs) stimulated with TNF (100 ng/ml), LPS (100 ng/ml), PAM 3 Cys (5 mcg/ml) for indicated times. G Correlation between hTNF and hIL-22BP levels in sera of IBD patients. All mouse data are representative of two independent experiments. Data represent mean values ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, as calculated by Student’s t-test; ns not significant. The Pearson correlation was used for correlative analyses. The significance level was set to p ≤ 0.05.

Article Snippet: Once mice had lost >5% of initial weight, they were treated with anti-human TNF antibody (Infliximab), anti-mouse IL-22 (R&D Systems; AF582) or human Fc-IgG1 isotype control (BioXCell; Cat # BE0096) in PBS.

Techniques: Recombinant, Derivative Assay, Expressing, Control, Isolation, Incubation

Journal: Mucosal Immunology

Article Title: TNF hampers intestinal tissue repair in colitis by restricting IL-22 bioavailability

doi: 10.1038/s41385-022-00506-x

Figure Lengend Snippet: A Levels of hTNF and mTNF in ex vivo colonic explants from Rag1 −/− mice following transfer of hTNF-KI T cells into Rag1 −/− mice and treated with Fc control or infliximab for 2 weeks. B Levels of hTNF and mTNF in ex vivo colonic explants from hTNF-KIxRag1 −/− mice that received T cells isolated from WT mice and were treated with Fc control or infliximab for 2 weeks. C hTNF and mTNF levels in sera of colitic Rag1 −/− mice transferred with hTNF-KI naive T cells before anti-TNF therapy. D Analysis of hTNF expression on the surface of blood T cells. Rag1 −/− mice were reconstituted with naive T cells from WT or hTNF-KI mice. Blood cells were analysed once mice had lost 5% of their initial weight. E Expression of IL-22BP in CD4 T cells isolated from WT and hTNF KI spleen. All data are representative of two independent experiments. Data represent mean values ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, as calculated by Student’s t-test; ns not significant.

Article Snippet: Once mice had lost >5% of initial weight, they were treated with anti-human TNF antibody (Infliximab), anti-mouse IL-22 (R&D Systems; AF582) or human Fc-IgG1 isotype control (BioXCell; Cat # BE0096) in PBS.

Techniques: Ex Vivo, Control, Isolation, Expressing

Journal: Mucosal Immunology

Article Title: TNF hampers intestinal tissue repair in colitis by restricting IL-22 bioavailability

doi: 10.1038/s41385-022-00506-x

Figure Lengend Snippet: A Weight of hTNFRp75‐KIxRag1 −/− mice reconstituted with naive T cells from hTNF‐KI mice and treated with Infliximab or Fc control (10 mg/kg) twice per week. IL-22 levels in colonic explants B and expression of IL-22BP mRNA in colon C of hTNFRp75‐KIxRag1 −/− mice reconstituted with naive T cells from hTNF-KI mice and treated with-infliximab for 2 weeks. Frequencies of CD11b + (CD45 + CD11b + ) D of granulocytes (CD45 + CD11b + Ly6G + Ly6C + ) and inflammatory monocytes (CD45 + CD11b + Ly6G - Ly6C + ) E , Th1 (CD45 + CD4 + TCRβ + IFNγ + ) F , Th17 (CD45 + CD4 + TCRβ + IL-17A + ) G and Th1/17 (CD45 + CD4 + TCRβ + IL-17A + IFNγ + ) H cells 2 weeks after infliximab or Fc control therapy. All data are representative of two independent experiments. Data represent mean values ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, as calculated by Student’s t-test; ns not significant.

Article Snippet: Once mice had lost >5% of initial weight, they were treated with anti-human TNF antibody (Infliximab), anti-mouse IL-22 (R&D Systems; AF582) or human Fc-IgG1 isotype control (BioXCell; Cat # BE0096) in PBS.

Techniques: Control, Expressing

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: HIV impairs TNF-alpha mediated macrophage apoptotic response to Mycobacterium tuberculosis.

doi: 10.4049/jimmunol.179.10.6973

Figure Lengend Snippet: FIGURE 2. TNF- release and the role of TNF- in MTb-mediated macrophage apoptosis. A, MTb-mediated macrophage apoptosis is dependent on TNF-. Differentiated U937 macrophages were incubated with MTb isolates in the presence and absence of specific neutralizing Ab to TNF- (anti-TNF; mAb 210, 5 g/ml) or isotype control (isotype), and after 24 h, apoptosis was determined by ELISA. Data are expressed as percent change in apoptosis compared with unstimulated macrophages (n 3). B, Reduced MTb-mediated TNF- release by HIV human macrophages. Differentiated human U937 and HIV U1 macrophages were incubated with individual MTb isolates at a multiplicity of 10:1 for 24 h, in the presence and absence of a general cellular phosphatase inhibitor, okadaic acid (OA; 50 ng/ml). Cell-free culture supernatants were assayed for TNF- by ELISA. C, Reduced TNF- mRNA transcripts in HIV macrophages. Differentiated U937 and HIV U1 macrophages were incubated for 6 h in the presence or absence of irradiated virulent MTb (iMTbRv). Specific TNF- mRNA was detected by RT-PCR, with -actin as an internal control for mRNA loading. Representative gel of three experiments providing identical results. D, Recovery of apoptosis in HIV macrophages by exogenous TNF- or endogenous macrophage TNF- release. Differentiated HIV U1 macrophages were incubated with irradiated virulent H37Rv MTb (iMTbRv) in the presence or absence of exogenous recombinant human TNF- (100 ng/ml), okadaic acid (OA; 50 ng/ml), or neutralizing anti-TNF Ab (mAb 210) for 24 h at 37°C. Apoptosis was determined by Cell Death ELISA. Data is shown as percentage change in OD from unstimulated condition. All quantitative data values represent mean SEM. , p 0.05 compared with iMTbRv alone; , p 0.05 compared with iMTbRv OA.

Article Snippet: Staurosporine (protein kinase C inhibitor; 5 M) was used as a positive control for apoptosis, and for select experiments, TNF- neutralizing Abs or isotype controls (mAb 210, R&D Systems) were added to culture supernatants to neutralize TNF- bioactivity before addition of MTb.

Techniques: Incubation, Control, Enzyme-linked Immunosorbent Assay, Irradiation, Reverse Transcription Polymerase Chain Reaction, Recombinant

Journal: Journal for Immunotherapy of Cancer

Article Title: Intratumoral CXCL13+ CD160+ CD8+ T cells promote the formation of tertiary lymphoid structures to enhance the efficacy of immunotherapy in advanced gastric cancer

doi: 10.1136/jitc-2024-009603

Figure Lengend Snippet: Targeting PDXK could promote the formation of TLSs and enhance the efficacy of immunotherapy in gastric cancer. ( A ) Heatmap displaying the metabolic feature for T-cell clusters using scMetabolism package. ( B ) Dot plot showing the expression of the genes encoding rate-limiting enzymes of vitamin B 6 metabolism in different types of T cells. Dot size encodes the percentage of cells expressing the gene, color encodes the average per cell gene expression level. ( C ) Quantification of CXCL13 in PDTFs in the presence of different enzyme inhibitors measured by ELISA. ( D ) Representative H&E staining and PDXK immunohistochemistry of gastric cancer tissues with different responses following immunotherapy. Scale bar, 500 µm. ( E ) The schematic diagram of the animal experiments. ( F–G ) Images of tumors and tumor volume curves of 615 mice treated with various agents (n=6, each group). ( H ) Paraffin sections of mouse subcutaneous graft tumor tissue stained with H&E and IHC detection for CD8, CD20 and CXCL13. Scale bar, 100 µm. ( I ) The number (left panel) and area (right panel) of TLS per tumor area were compared between groups (n=6, each group). Data are presented as the mean±SD. ns, not significant. *p<0.05, ***p<0.001, two-tailed Student’s t-test. AOX1, aldehyde oxidase 1; CCCP, carbonyl cyanide m-chlorophenyl hydrazone; CR, complete response; MFC, mouse forestomach carcinoma; PBS, phosphate-buffered saline; PDXK, pyridoxal kinase; PDXP, pyridoxal phosphatase; PDTFs, patient-derived tumor fragments; PHOSPHO2, phosphatase orphan 2; PNPO, pyridoxamine 5'-phosphate oxidase; PR, partial response; PSAT1, phosphoserine aminotransferase 1; s.c, subcutaneous injections; SD, stable disease; TLS, tertiary lymphoid structures.

Article Snippet: The indicated cytokines and chemokines within the supernatants were detected using human IL-2 ELISA Kit (Solarbio, SEKH-0008), human IFN-γ ELISA Kit (Solarbio, SEKH-0046), human CXCL13 ELISA Kit (Solarbio, SEKH-0072) and human TNF-α ELISA Kit (Solarbio, SEKH-0047) according to the manufacturers’ instructions.

Techniques: Expressing, Gene Expression, Enzyme-linked Immunosorbent Assay, Staining, Immunohistochemistry, Two Tailed Test, Saline, Derivative Assay

Journal: Journal for Immunotherapy of Cancer

Article Title: Intratumoral CXCL13+ CD160+ CD8+ T cells promote the formation of tertiary lymphoid structures to enhance the efficacy of immunotherapy in advanced gastric cancer

doi: 10.1136/jitc-2024-009603

Figure Lengend Snippet: Vitamin B 6 could promote the expression and secretion of CXCL13 in CD160 + CD8 + T cells. ( A ) Quantification of cytokine/chemokine including CXCL13, IL-2, TNF-α and IFN-γ, in PDTFs in the presence of different drugs measured by ELISA. ( B ) The schematic diagram of orthotopic transplanted tumor model with diets containing various amounts of vitamin B 6 (n=6, each group). ( C ) Mouse orthotopic stomach xenograft tumor tissues stained with H&E and IHC detection for CD20 and CXCL13. Scale bar, 500 µm. ( D ) The schematic diagram of orthotopic transplanted tumor model fed with different drugs or diets (n=6, each group). ( E ) The representative images of mouse bioluminescence imaging at week 3 (left panel) and the corresponding quantification analysis (right panel). ( F ) Representative micrographs of xenografts stained with H&E and IHC detection for CD20 and CXCL13. Scale bar, 500 µm. ( G–H ) Density of TLSs (left panel) and ratio of tumor area occupied by TLSs (right panel). ( I ) Gating strategy for CD160 + CD8 + T cells. ( J ) Quantification of cytokine/chemokine including CXCL13, IL-2, TNF-α and IFN-γ, in supernatants from CD160 + CD8 + T-cell cultures in the presence or absence of PL measured by ELISA. ( K ) Flow cytometric analysis and corresponding quantification of CXCL13 + CD160 + CD8 + T cells with or without PL treatment. Data are presented as the mean±SD. ns, not significant. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, two-tailed Student’s t-test. ICIs, immune checkpoint inhibitors; IFN, interferon; IHC, immunohistochemistry; IL, interleukin; MFC, mouse forestomach carcinoma; PBS, phosphate-buffered saline; PL, pyridoxol; PD-1, programmed cell death protein 1; s.c, subcutaneous injections; TLS, tertiary lymphoid structures; TNF, tumor necrosis factor.

Article Snippet: The indicated cytokines and chemokines within the supernatants were detected using human IL-2 ELISA Kit (Solarbio, SEKH-0008), human IFN-γ ELISA Kit (Solarbio, SEKH-0046), human CXCL13 ELISA Kit (Solarbio, SEKH-0072) and human TNF-α ELISA Kit (Solarbio, SEKH-0047) according to the manufacturers’ instructions.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Staining, Imaging, Two Tailed Test, Immunohistochemistry, Saline

Journal: Molecules

Article Title: Phenolic and Non-Polar Fractions of the Extracts from Fruits, Leaves, and Twigs of Elaeagnus rhamnoides (L.) A. Nelson—The Implications for Human Barrier Cells

doi: 10.3390/molecules25092238

Figure Lengend Snippet: Figure 1. IL-8 production by human colon epithelial-like cells line Caco-2 exposed to fractionated Elaeagnus rhamnoides (L.) extracts and reference compounds: (a) 6 h; (b) 24 h, tested by ELISA. The results are given as a mean concentration (pg/mL) ± standard error. C1—control cells in culture medium with Me-OH, OF/GF/LF—phenolic fractions of fruit/twig/leaf extracts, C2—control cells in culture medium with DMSO, OL/GL/LL—non-polar fractions of fruit/twig/leaf extracts, PG—Staphylococcus aureus peptidoglycan, TNF—murine tumor necrosis factor alpha, KU—ursolic acid, Q—quercetin, E—(−)-epicatechin, KE—ellagic acid; * p < 0.05, ** p < 0.005.

Article Snippet: Molecules 2020, 25, 2238 14 of 19 Human IL-8/CXCL8 DuoSet ELISA, human TNF-α DuoSet ELISA, murine TNF-α (TNF), murine monoclonal anti-human CCL-3/MIP-1 DuoSet ELISA, human ICAM-1/CD54 fluorescein-conjugated antibodies, rat monoclonal anti-human/mouse/bovine integrin alpha 6/CD49f phycoerythrin-conjugated antibodies and murine monoclonal IgG1 fluorescein-conjugated antibodies were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay, Control

Journal: Molecules

Article Title: Phenolic and Non-Polar Fractions of the Extracts from Fruits, Leaves, and Twigs of Elaeagnus rhamnoides (L.) A. Nelson—The Implications for Human Barrier Cells

doi: 10.3390/molecules25092238

Figure Lengend Snippet: Figure 3. IL-8 production by human foreskin fibroblasts line HFF-1 exposed to fractionated E. rhamnoides (L.) extracts and reference compounds: (a) 6 h; (b) 24 h, tested by ELISA. The results are given as a mean concentration [pg/mL] ± standard error. C1—control cells in culture medium with Me-OH, OF/GF/LF—phenolic fractions of fruit/twig/leaf extracts, C2—control cells in culture medium with DMSO, OL/GL/LL—non-polar fractions of fruit/twig/leaf extracts, PG—S. aureus peptidoglycan, TNF—murine tumor necrosis factor alpha, KU—ursolic acid, Q—quercetin, E—(−)-epicatechin, KE—ellagic acid; * p < 0.05, ** p < 0.01.

Article Snippet: Molecules 2020, 25, 2238 14 of 19 Human IL-8/CXCL8 DuoSet ELISA, human TNF-α DuoSet ELISA, murine TNF-α (TNF), murine monoclonal anti-human CCL-3/MIP-1 DuoSet ELISA, human ICAM-1/CD54 fluorescein-conjugated antibodies, rat monoclonal anti-human/mouse/bovine integrin alpha 6/CD49f phycoerythrin-conjugated antibodies and murine monoclonal IgG1 fluorescein-conjugated antibodies were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay, Control