Journal: Clinical and Experimental Immunology

Article Title: Investigating the role of proinflammatory CD16 + monocytes in the pathogenesis of inflammatory bowel disease

doi: 10.1111/j.1365-2249.2010.04177.x

Figure Lengend Snippet: The peripheral blood CD14+ CD16+ monocyte population is increased in active Crohn's disease. (a) Peripheral blood mononuclear cells (PBMC) were stained for CD14, CD16 and CD36 and analysed by flow cytometry. Cells were pre-gated for high forward-scatter (FSC), side-scatter (SSC) and CD36 expression, and monocytes separated by CD14/CD16 expression. The lower panels show representative dot plots from a healthy volunteer, and Crohn's disease patients with quiescent [Crohn's disease activity index (CDAI)<150] and active (CDAI>150) inflammation. (b) The ratio of CD14+ CD16+ monocytes was increased significantly in Crohn's disease patients compared to healthy controls. *P = 0·01 (two-tailed Student's t-test with Welch's correction). (c) Further categorization of patients revealed that the CD14+ CD16+ population was increased in active Crohn's disease compared to healthy controls, but not quiescent disease or patients with ulcerative colitis. *P < 0·05 (Bonferroni's post-test following one-way analysis of variance).

Article Snippet: Slides were then incubated at 4°C overnight with the following primary antibodies: mouse anti-CD14 (BioLegend, San Diego, CA, USA), rat anti-CD16 (Santa Cruz Biotechnology, Santa Cruz, CA, USA) and rabbit anti-CD36 (Novus Biologicals, Littleton, CO, USA).

Techniques: Staining, Flow Cytometry, Expressing, Activity Assay, Two Tailed Test

Journal: Clinical and Experimental Immunology

Article Title: Investigating the role of proinflammatory CD16 + monocytes in the pathogenesis of inflammatory bowel disease

doi: 10.1111/j.1365-2249.2010.04177.x

Figure Lengend Snippet: The number of CD14+ CD16+ monocytes is enhanced significantly in the lamina propria of Crohn's disease patients. (a) Monocytic lineage of CD14+ CD16+ cells in the colonic mucosa was confirmed by three-channel immunofluorescence microscopy for CD14 (green), CD16 (red) and CD36 (blue). No or limited CD36 staining was observed in CD14+ CD16– cells (i) and CD14– CD16+ cells (ii). In contrast, CD14+ CD16+ cells stained strongly for CD36 (iii, white pseudocolour). Colonic crypts are indicated with dashed lines. Lower panels, magnification of the boxed cells shown above. (b) Representative images of colonic mucosal sections from non-inflammatory bowel disease (IBD) control patients, non-inflamed and inflamed tissue from Crohn's disease patients, and non-inflamed ulcerative colitis samples. CD14+ CD16+ monocytes are shown in yellow pseudocolour, with nuclei in blue. (c) The number of CD14+ CD16+ cells per high-power field (HPF) was significantly increased in Crohn's disease mucosa, and enhanced further in actively inflamed tissue. Each data point represents the average of five or more HPF per patient sample. *P < 0·001 versus control, †P < 0·001 versus IBD – not inflamed [Bonferroni's post-test following one-way analysis of variance (anova)]. (d) The ratio of CD14+ CD16+ cells was increased significantly in Crohn's disease patients compared to controls. *P < 0·001 (Bonferroni's post-test following one-way anova). (e) The increase in total CD14+ CD16+ monocytes was confirmed by flow cytometry. Mononuclear cells were isolated from control and Crohn's disease colonic mucosa as described in the Materials and methods, pre-gated for forward-scatter (high), side-scatter (high) CD36+, and separated by CD14/CD16 expression. A dramatic increase of CD14+ CD16+ monocytes was seen in IBD tissue (red box). Plots are representative of three patient samples per group. (f) The tumour necrosis factor (TNF)-α mean fluorescence intensity (MFI, arbitrary units) of CD14+ CD16+ monocytes was significantly higher compared to CD14+ CD16– cells in the same field of vision. The graph shows the result from 14 HPF taken from three Crohn's disease patient samples. *P < 0·001 (two-tailed Student's t-test).

Article Snippet: Slides were then incubated at 4°C overnight with the following primary antibodies: mouse anti-CD14 (BioLegend, San Diego, CA, USA), rat anti-CD16 (Santa Cruz Biotechnology, Santa Cruz, CA, USA) and rabbit anti-CD36 (Novus Biologicals, Littleton, CO, USA).

Techniques: Immunofluorescence, Microscopy, Staining, Flow Cytometry, Isolation, Expressing, Fluorescence, Two Tailed Test

Journal: Oral diseases

Article Title: CD36 Is Upregulated in Mice with Periodontitis and Metabolic Syndrome and Involved in Macrophage Gene Upregulation by Palmitate

doi: 10.1111/odi.12596

Figure Lengend Snippet: The periodontal CD36 protein expression in mice without MetS and periodontitis, with MetS or periodontitis alone, and with both MetS and periodontitis. C57BL/6 mice with or without HFD-induced MetS were injected with PBS or LPS in periodontal tissue. After the treatment, CD36 protein expression was determined by immunohistochemistry as described in Methods. Representative images of CD36 immunostaining in the subepithelial tissue were shown (A). The areas with positive CD36 staining were quantified (B) (n=7).

Article Snippet: The sections were then incubated with 5% normal goat serum in 0.01 M PBS for 1 hour to block nonspecific binding and incubated with rabbit anti-CD36 antibody (1:500) (Novus biological, Littleton, CO) overnight at 4°C.

Techniques: Expressing, Injection, Immunohistochemistry, Immunostaining, Staining

Journal: Oral diseases

Article Title: CD36 Is Upregulated in Mice with Periodontitis and Metabolic Syndrome and Involved in Macrophage Gene Upregulation by Palmitate

doi: 10.1111/odi.12596

Figure Lengend Snippet: The periodontal CD36 mRNA expression in mice without MetS and periodontitis, with MetS or periodontitis alone, and with both MetS and periodontitis. C57BL/6 mice with or without HFD-induced MetS were injected with PBS or LPS in periodontal tissue. After the treatment, RNA was isolated from the periodontal tissues and CD36 mRNA expression was quantified by real-time PCR as described in Methods. The data presented are mean ± SD (n=7).

Article Snippet: The sections were then incubated with 5% normal goat serum in 0.01 M PBS for 1 hour to block nonspecific binding and incubated with rabbit anti-CD36 antibody (1:500) (Novus biological, Littleton, CO) overnight at 4°C.

Techniques: Expressing, Injection, Isolation, Real-time Polymerase Chain Reaction

Journal: Oral diseases

Article Title: CD36 Is Upregulated in Mice with Periodontitis and Metabolic Syndrome and Involved in Macrophage Gene Upregulation by Palmitate

doi: 10.1111/odi.12596

Figure Lengend Snippet: The CD36 mRNA expression in RAW264.7 macrophages treated with LPS, palmitate or LPS plus palmitate. RAW264.7 macrophages were treated with 1 ng/ml of LPS, 100 μM of palmitate or LPS plus palmitate for 24 h and CD36 mRNA was then quantified using real-time PCR. The data presented are mean ± SD of three experiments.

Article Snippet: The sections were then incubated with 5% normal goat serum in 0.01 M PBS for 1 hour to block nonspecific binding and incubated with rabbit anti-CD36 antibody (1:500) (Novus biological, Littleton, CO) overnight at 4°C.

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Oral diseases

Article Title: CD36 Is Upregulated in Mice with Periodontitis and Metabolic Syndrome and Involved in Macrophage Gene Upregulation by Palmitate

doi: 10.1111/odi.12596

Figure Lengend Snippet: Correlation between periodontal CD36 protein expression and osteoclast formation and alveolar bone volume fraction (BVF). C57BL/6 mice with or without HFD-induced MetS were injected with PBS or LPS in periodontal tissue to induce periodontitis. After the treatment, CD36 protein expression was quantified using immunohistochemistry, osteoclasts were detected using tartrate-resistance acid phosphatase (TRAP) staining, and alveolar BVF were quantified using μCT. The correlations between CD36 expression and osteoclast formation (A) or alveolar BVF (B) were statistically analyzed.

Article Snippet: The sections were then incubated with 5% normal goat serum in 0.01 M PBS for 1 hour to block nonspecific binding and incubated with rabbit anti-CD36 antibody (1:500) (Novus biological, Littleton, CO) overnight at 4°C.

Techniques: Expressing, Injection, Immunohistochemistry, Staining

Journal: Oral diseases

Article Title: CD36 Is Upregulated in Mice with Periodontitis and Metabolic Syndrome and Involved in Macrophage Gene Upregulation by Palmitate

doi: 10.1111/odi.12596

Figure Lengend Snippet: The inhibitory effect of CD36 inhibitor SSO on the enhancement by palmitate of LPS-induced IL-6 secretion. RAW264.7 macrophages were treated with 1 ng/ml of LPS, 100 μM of palmitate or LPS plus palmitate in the absence or presence of 100 μM of SSO for 24 h and IL-6 secreted into culture medium was then quantified using ELISA. The data presented are mean ± SD of three experiments.

Article Snippet: The sections were then incubated with 5% normal goat serum in 0.01 M PBS for 1 hour to block nonspecific binding and incubated with rabbit anti-CD36 antibody (1:500) (Novus biological, Littleton, CO) overnight at 4°C.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Oral diseases

Article Title: CD36 Is Upregulated in Mice with Periodontitis and Metabolic Syndrome and Involved in Macrophage Gene Upregulation by Palmitate

doi: 10.1111/odi.12596

Figure Lengend Snippet: The inhibitory effect of CD36 knockdown on the enhancement by palmitate of LPS-induced IL-6 expression and secretion. RAW264.7 macrophages were transfected with 10 nM control or CD36 siRNA for 48 h and then treated with 1 ng/ml of LPS, 100 μM of palmitate or both LPS and palmitate for 24 h. After the treatment, IL-6 mRNA expression and secretion was quantified using ELISA. A: CD36 expression in cells after transfection with control or CD36 siRNA. B. IL-6 mRNA expression in cells treated with LPS, palmitate or LPS plus palmitate after transfection with either control or CD36 siRNA. C. IL-6 secretion by cells treated with LPS, palmitate or LPS plus palmitate after transfection with either control or CD36 siRNA. The data presented are mean ± SD of three experiments.

Article Snippet: The sections were then incubated with 5% normal goat serum in 0.01 M PBS for 1 hour to block nonspecific binding and incubated with rabbit anti-CD36 antibody (1:500) (Novus biological, Littleton, CO) overnight at 4°C.

Techniques: Knockdown, Expressing, Transfection, Control, Enzyme-linked Immunosorbent Assay

Journal: Oral diseases

Article Title: CD36 Is Upregulated in Mice with Periodontitis and Metabolic Syndrome and Involved in Macrophage Gene Upregulation by Palmitate

doi: 10.1111/odi.12596

Figure Lengend Snippet: The effect of CD36 inhibitor SSO on the expression of pro-inflammatory molecules stimulated by LPS, palmitate or LPS in combination with palmitate

Article Snippet: The sections were then incubated with 5% normal goat serum in 0.01 M PBS for 1 hour to block nonspecific binding and incubated with rabbit anti-CD36 antibody (1:500) (Novus biological, Littleton, CO) overnight at 4°C.

Techniques: Expressing, Inhibition

Journal: Cancer & Metabolism

Article Title: Integration of global metabolomics and lipidomics approaches reveals the molecular mechanisms and the potential biomarkers for postoperative recurrence in early-stage cholangiocarcinoma

doi: 10.1186/s40170-021-00266-5

Figure Lengend Snippet: The summary of pathway analysis on differential metabolites between recurrence and non-recurrence, analyzed using MetaboAnalyst 4.0. A Metabolism pathway analysis from global metabolomics. B Metabolism pathway analysis from lipidomics. The color of the circle represents the p value, and the size of the circle represents the pathway impact. C The schematic diagram of metabolic pathways involved in CCA recurrence with red arrows indicating the most relevant pathway for recurrence. FAO fatty acid oxidation, CSC cancer stem cell, TG triacylglycerol, NEAAs non-essential amino acids, CD36 cluster of differentiation 36, ACLY ATP citrate lyase, SCD1 stearoyl-CoA desaturase-1

Article Snippet: The antibodies used in this study were mouse monoclonal anti-CD44 (1:100; #ab516728), mouse monoclonal anti-CD44v6 (1:50; #ab78960), rabbit polyclonal anti-EpCAM (1:100; #ab71916), rabbit monoclonal anti-CD36 (1:25; #ab133625), rabbit monoclonal anti-ATP citrate lyase (1:200; #ab40793), rabbit monoclonal anti-SCD1 (1:100; #ab236868) and HRP-conjugated rabbit anti-rat (1:50; #ab6734) antibodies (Abcam, CA), and rat monoclonal anti-CD44v8-10 antibody (1:50; #LKG-M001) (Cosmo Bio, JP).

Techniques:

Journal: PLoS ONE

Article Title: Oxidation by Neutrophils-Derived HOCl Increases Immunogenicity of Proteins by Converting Them into Ligands of Several Endocytic Receptors Involved in Antigen Uptake by Dendritic Cells and Macrophages

doi: 10.1371/journal.pone.0123293

Figure Lengend Snippet: ( A ) Uptake of fluorescently-labelled proteins by CHO cells transfected with human CD36 as compared to non-transfected cells. ( B ) Binding of anti-CD36 mAb and control mouse IgA to CD36-transfected and non-transfected CHO cells, determined by cellular ELISA. ( C ) Binding of rCD36 to plate-adsorbed proteins. ( D ) Effects of 10 or 100 μg/ml of indicated, soluble ligands on rCD36 binding to plate-adsorbed OVA-Cl. ( E ) Binding of polyclonal anti-mouse CD36 Ab and control goat IgG to BM-DC, splenic DC and PEM. A representative histogram of Ab binding to BM-DC is displayed on the left graph. ( F ) Binding of anti-mouse SR-A mAb and control rat IgG2b to BM-DC, splenic DC and PEM. A representative histogram of Ab binding to BM-DC is shown on the left graph. ( G ) Binding of SR-A present in lysates of PEM to plate-adsorbed proteins. ( H ) Effects of anti-SR-A 2F8 mAb and AcLDL on the uptake of fluorescently-labelled proteins by BM-DC. Shown are results of single experiments, each representative of at least 3 similar experiments performed (A-D, G, H) or averages +SEM from 4–6 independent experiments (E, F). The data were analysed with the unpaired (A-C, G) or one-sample (H) Student’s t-test or with ANOVA, followed by the Tukey-Kramer post-test (D). *, p < 0.05; ND, not done.

Article Snippet: Rat anti-mouse scavenger receptor A (SR-A) mAb (clone 2F8) was obtained from AbD Serotec; mouse anti-mouse CD36 mAb (CRF D-2712) from Hycult Biotech; mouse IgA isotype control mAb (M18-254), rat IgG2b isotype control mAb (A95-1), rat anti-mouse CD11b mAb (M1/70) and phycoerythrin (PE)-streptavidin conjugate from BD Biosciences; rat IgG2a isotype control mAb (54447), normal goat IgG, polyclonal goat anti-mouse CD36, anti-mouse LOX-1 (lectin-type oxidised LDL receptor-1), anti-human SREC-I (scavenger receptor expressed by endothelial cells-I) Ab and PE-conjugated rat anti-mouse LOX-1 mAb (214012) from R&D Systems; polyclonal goat anti-mouse SREC-I, anti-mouse RAGE (receptor for advanced glycation end products), anti-mouse stabilin-1 and rabbit anti-mouse-stabilin-1 Ab from Santa Cruz Biotechnology; PE-conjugated donkey anti-goat IgG Ab from SouthernBiotech; rat anti-mouse CD206/mannose receptor mAb (MR5D3) and PE-conjugated goat anti-rat IgG Ab from BioLegend; horseradish peroxidase (HRP)-conjugated rabbit anti-mouse IgA, F(ab’)2 fragments of goat anti-rat IgG and donkey anti-goat IgG Ab from Rockland.

Techniques: Transfection, Binding Assay, Enzyme-linked Immunosorbent Assay

Journal: Cell Reports Medicine

Article Title: TRPM2 overactivation drives hyperlipidemia-induced dysfunction of myeloid cells and neurovascular units

doi: 10.1016/j.xcrm.2025.101998

Figure Lengend Snippet: Global TRPM2 knockout abolishes the exacerbation of ischemic brain injury by hyperlipidemia (A) The presence of atherosclerotic plaque (indicated by black arrows) at the bifurcation of common carotid artery (CCA) and in the internal carotid artery (ICA) (scale bar size: 2 mm). (B–E) Triphenyl tetrazolium chloride (TTC) staining 24 h after MCAO (60-min) and neurological deficit score in WT, Apoe −/− mice, and Apoe −/− gM2KO mice (B and C) and in Trpm2 fl/fl mice, Apoe −/− Trpm2 fl/fl cre − mice, and Apoe −/− Trpm2 fl/fl cre + mice (D and E) fed with or without HFD ( n = 12–20 mice per group) (scale bar size: 5 mm). (F and G) TRPM2 expression in brains from WT mice fed with or without HFD ( n = 5 and 5 mice). (H and I) TRPM2 and CD36 expression in primary neurons, CECs, BMDMs, and peripheral leukocytes isolated from WT mice ( n = 5, 5, 5, and 5 dishes of cells isolated from at least 5 mice). (J and K) TRPM2 expression in peripheral leukocytes isolated from WT mice fed with HFD for 0, 1, 2, 3, and 4 months ( n = 5, 5, 5, 5, and 5 dishes of cells isolated from at least 5 mice). (L) ELISA measurement of TRPM2 levels in lysates from B cells, T cells, monocytes, and neutrophils after cell sorting of peripheral leukocytes from WT mice with or without HFD treatment. (M and N) ELISA measurement of plasma oxLDL and IL-1β levels in WT mice fed with HFD for 0, 1, 2, 3, and 4 months ( n = 5, 5, 5, 5, and 5 plasma samples from different mice). (O and P) Correlation of plasma IL-1β level with plasma oxLDL level (O) and TRPM2 expression (P) in peripheral leukocytes ( n = 5 mice). (Q) Correlation of plasma oxLDL level with TRPM2 expression in peripheral leukocytes ( n = 5 mice). Error bars: mean ± SEM; ns, no statistical significance, ∗, p < 0.05, ∗∗, p < 0.01, ∗∗∗, p < 0.001. See also Figure S2 and Table S1 .

Article Snippet: CD36 (D8L9T) Rabbit mAb , Cell Signaling Technology , Cat#14347; RRID: AB_2798555.

Techniques: Knock-Out, Staining, Expressing, Isolation, Enzyme-linked Immunosorbent Assay, FACS, Clinical Proteomics

Journal: Cell Reports Medicine

Article Title: TRPM2 overactivation drives hyperlipidemia-induced dysfunction of myeloid cells and neurovascular units

doi: 10.1016/j.xcrm.2025.101998

Figure Lengend Snippet: Increase of TRPM2 expression by oxLDL compromises resistance of endothelial cells to ischemia (A–C and E) TRPM2 and pp65 expression in primary CECs isolated from WT mouse treated with oxLDL (A and B) and LDL (C and E) for 0, 6, 12, 18, 24, and 48 h ( n = 5). (D and F) CD36 expression in primary CECs isolated from WT mouse treated with oxLDL for 0, 6, 12, 18, 24, and 48 h ( n = 5). (G and H) TRPM2 expression in WT mouse primary CECs treated with oxLDL for 48 h with the co-treatment of DMSO, SSO, and scavengers Mn (III) TBAP and L-NMA ( n = 5). (I and J) TRPM2, iNOS, and pp65 expression in WT and M2KO mouse primary CECs treated with oxLDL for 48 h with the co-treatment of DMSO, SN50, ACA, and TAT-M2 ( n = 5). (K and L) Examination of endothelial responses to in vitro ischemia insult. TRPM2, iNOS, pp65, and occludin expression in WT and M2KO mouse primary CECs subjected to OGD for 8 h ( n = 5). (M and N) Monitoring of the loss of giga-seal after OGD in transwell inserts plated with WT or M2KO mouse primary CECs pretreated with oxLDL for 48 h ( n = 3). (O) Measurement of the leakage of Evans blue (related to the PBS-Con groups) from upper into lower chamber 8 h after OGD in transwell inserts ( n = 6). n means X dishes of cells isolated from at least X mice per group; error bars: mean ± SEM; ns, no statistical significance, ∗, p < 0.05, ∗∗, p < 0.01, ∗∗∗, p < 0.001.

Article Snippet: CD36 (D8L9T) Rabbit mAb , Cell Signaling Technology , Cat#14347; RRID: AB_2798555.

Techniques: Expressing, Isolation, In Vitro

Journal: Cell Reports Medicine

Article Title: TRPM2 overactivation drives hyperlipidemia-induced dysfunction of myeloid cells and neurovascular units

doi: 10.1016/j.xcrm.2025.101998

Figure Lengend Snippet:

Article Snippet: CD36 (D8L9T) Rabbit mAb , Cell Signaling Technology , Cat#14347; RRID: AB_2798555.

Techniques: Recombinant, Clinical Proteomics, Sequencing, Enzyme-linked Immunosorbent Assay, Bicinchoninic Acid Protein Assay, Knock-Out, Generated, Software, Microscopy, Imaging