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: Small (Weinheim an der Bergstrasse, Germany)

Article Title: Probiomimetics-Novel Lactobacillus-Mimicking Microparticles Show Anti-Inflammatory and Barrier-Protecting Effects in Gastrointestinal Models.

doi: 10.1002/smll.202003158

Figure Lengend Snippet: Figure 6. Cytokine production in inflamed macrophage-like dTHP-1 cells as well as in enterocyte-like Caco-2 cells. Cells were stimulated with lipopoly- saccharides (10 µg mL-1) and cotreated with probiomimetics or MVs. Supernatants were harvested after 6 or 24 h, and the protein content was analyzed using ELISA. To exclude the effects of different MV concentrations during the assembly of the probiomimetics, all results were normalized to the protein content. a) ELISA measurement of proinflammatory TNF-α released in dTHP-1 cells after 6 h. b) Release of anti-inflammatory IL-10 after 6 h. c) ELISA measurement of proinflammatory TNF-α released in dTHP-1 cells after 24 h. d) Release of anti-inflammatory IL-10 after 24 h. e) Release of proinflam- matory IL-8 in enterocyte-like Caco-2 cells after 24 h. Values represent the mean of 3–9 biological replicates with standard deviations.

Article Snippet: The supernatants were thawed and the concentrations of IL-10 and TNF-α were analyzed using Human IL-10 ELISA Set (Diaclone, Besançon, France), Human TNF-α ELISA Set (Diaclone, Besançon, France), and Human IL-8 ELISA Set (Diaclone, Besançon, France), respectively, according to the supplier’s protocols.

Techniques: Enzyme-linked Immunosorbent 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: 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