Journal: Journal of Advanced Research

Article Title: Novel function of macrophage migration inhibitory factor in regulating post-infarct inflammation and the therapeutic significance

doi: 10.1016/j.jare.2025.05.030

Figure Lengend Snippet: Macrophage migration inhibitory factor (MIF) promotes inflammatory cell migration and infiltration through C – C motif chemokine receptor 2 (CCR2) and C-X-C motif chemokine receptor (CXCR) 4 and promotes splenic monocyte mobilization via interaction with angiotensin Ⅱ type 1 receptor (AT-1R) after myocardial infarction (MI). A, peripheral blood mononuclear cells (PBMCs) chemotaxis in response to homogenized sham or infarct heart tissue from wild type (WT) mice at day 3 post-MI, or MI homogenate together with inhibition of C–C motif chemokine receptor 2 (CCR2), C-X-C motif chemokine receptor (CXCR) 2 and CXCR4, respectively. Cells/HPF, cells/high power field. PBMCs harvested from WT mice at 3 days post-MI. CCR2i, CCR2 inhibition. CXCR2i, CXCR2 inhibition. CXCR4i, CXCR4 inhibition. n = 4–8 per group. *** P < 0.001 vs. sham . B, PBMCs chemotaxis in response to recombinant human MIF (rMIF) or rMIF with inhibition of CCR2, CXCR2 and CXCR4, respectively. PBMCs harvested from WT mice at 3 days post-MI. n = 4–8 per group. *** P < 0.001 vs. control . C, Representative immunoblotting images for monocyte chemoattractant protein-1 (MCP-1), CXCR4 and internal reference protein (heat shock protein 60, HSP-60) in hearts from WT and global MIF deficient (MIFKO) mice with sham-operation or 72 h MI. D-E, Quantitative analysis of MCP-1 and CXCR4 expression. n = 6–9 per group. ** P < 0.01 vs. sham in the same genotype. *** P < 0.001 vs. sham in the same genotype. F, Splenic monocyte (from normal WT mice) chemotaxis in response to plasma from normal mice or mice with 1.5 h MI, or MI plasma together with losartan. Lor, losartan, AT-1R inhibitor. n = 6–8 per group. *** P < 0.001 vs. normal. G, Splenic monocytes (from normal WT mice) chemotaxis in response to rMIF and angiotensin II (Ang II), or together with losartan addition, respectively. n = 6–8 per group. ** P < 0.01 vs. Control. *** P < 0.001 vs. Control. H, Representative immunoblotting images for AT-1R and internal reference protein (HSP-60) in spleens from WT and global MIF deficient (MIFKO) mice with sham-operation and 1.5 h MI. I, Quantitative analysis of AT-1R expression. n = 8 per group. *** P < 0.001 vs. sham in the same genotype . J. Co-immunoprecipitation (Co-IP) assay using an anti-AT-1R antibody or IgG demonstrated the interaction between AT-1R and MIF, as well as between AT-1R and CD74 in splenic monocytes and spleen tissue. Input, input sample. IgG, negative control sample. IP, Co-IP sample.

Article Snippet: For both groups 1 and 2, PBMCs were pre-treated for 1 h using DMEM with or without CCR2 inhibitor (CCR2 antagonist 4 hydrochloride, 100 mM), CXCR2 inhibitor (SB225002, 800 nM) or CXCR4 inhibitor (Plerixafor, AMD3100, 10 μM) (all from MedChemExpress, USA).

Techniques: Migration, Chemotaxis Assay, Inhibition, Recombinant, Control, Western Blot, Expressing, Clinical Proteomics, Co-Immunoprecipitation Assay, Negative Control

Journal: Inflammation

Article Title: Activation of β2-Adrenergic Receptor Alleviates Viral Myocarditis by Regulating Energy Metabolism in Monocyte-derived Macrophages via the AMPK Pathway

doi: 10.1007/s10753-025-02382-6

Figure Lengend Snippet: Activating β2-AR attenuates inflammation and fibrosis in VMC. A - B Representative flow cytometry images (A) and statistical results (B) of β2-AR expression on CD45 + CD64 + F4/80 + CCR2 + macrophages in control and myocarditis model mice at 1 week and 2 weeks post-CVB3 infection ( n = 6/group). C - D Representative H&E-stained images of myocardial tissue from each group at 1 week (C) and 2 weeks (D) post-CVB3 infection (original magnification, 200 × ; scale bars = 50 µm). E – F Representative Masson's trichrome staining images from each group at 1 week (E) and 2 weeks (F) post-CVB3 infection (original magnification, 200 × ; scale bars = 50 µm). G - H Representative cardiac ultrasound images of the mice in each group at 1 week (G) and 2 weeks (H) post-CVB3 infection. (I) Statistical analysis of the myocardial pathological scores for each group at 1 week and 2 weeks post-CVB3 infection ( n = 6/group). J Statistical analysis of cardiac collagen volume fractions in each group at 1 week and 2 weeks post-CVB3 infection ( n = 6/group). K - L Statistical analysis of the LVFS (K) and LVEF (L) of each group at 1 week and 2 weeks post-CVB3 infection ( n = 6/group). M – O Real-time qPCR analysis of CCL2 (M), TNF-α (N), and IL-10 (O) expression in each group at 1 week and 2 weeks post-CVB3 infection ( n = 6/group). The data are presented as the means ± SEMs. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns = not significant. Abbreviations: CCL2, C–C motif chemokine ligand 2; CVB3, coxsackievirus B3; H&E, hematoxylin and eosin; LVFS, left ventricular fractional shortening; LVEF, left ventricular ejection fraction; IL-10, interleukin 10; qPCR, real-time quantitative polymerase chain reaction; SEM, standard error of the mean; TNF-α, tumor necrosis factor-α; VMC, viral myocarditis; β2-adrenergic receptor, β2-AR

Article Snippet: Following established protocols [ ], the C–C motif chemokine receptor 2 (CCR2) inhibitor INCB3344 (HY-50674, MCE, USA) was delivered intraperitoneally at 30 mg/kg/day to VMC mice, starting on Day 1 and continuing until Day 14 post-CVB3 infection.

Techniques: Flow Cytometry, Expressing, Control, Infection, Staining, Real-time Polymerase Chain Reaction

Journal: Inflammation

Article Title: Activation of β2-Adrenergic Receptor Alleviates Viral Myocarditis by Regulating Energy Metabolism in Monocyte-derived Macrophages via the AMPK Pathway

doi: 10.1007/s10753-025-02382-6

Figure Lengend Snippet: Effects of activating β2-AR on the frequencies and subsets of monocytes/macrophages. A Representative flow cytometry images of cardiac macrophage subsets among CD45 + CD64 + F4/80 + cells in the control, CVB3 + vehicle, and CVB3 + formoterol-treated groups at 1 week and 2 weeks post-infection. B - C Statistical analysis of CCR2 + macrophages (B) and CCR2 − macrophages (C) among cardiac macrophages in the control, CVB3 + vehicle, and CVB3 + formoterol-treated groups at 1 week and 2 weeks post-infection ( n = 6/group). D Representative flow cytometry images of Ly6C high and Ly6C low monocytes among blood CD45 + CD11b + Ly6G − cells in the control, CVB3 + vehicle, and CVB3 + formoterol-treated groups at 1 week and 2 weeks post-infection. E – F Statistical analysis of Ly6C high monocytes (E) and Ly6C low monocytes (F) among blood CD45 + CD11b + Ly6G − cells in the control, CVB3 + vehicle, and CVB3 + formoterol-treated groups at 1 week and 2 weeks post-infection ( n = 6/group). G - H Representative flow cytometry images (G) and statistical analysis (H) of Ly6C + monocytes among splenic CD45 + CD11b + Ly6G − cells in the control, CVB3 + vehicle, and CVB3 + formoterol-treated groups at 1 week and 2 weeks post-infection (n = 6/group). The data are presented as the means ± SEMs. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns = not significant. Abbreviations: CCR2, C-C chemokine receptor type 2; CVB3, coxsackievirus B3; SEM, standard error of the mean; β2-adrenergic receptor, β2-AR

Article Snippet: Following established protocols [ ], the C–C motif chemokine receptor 2 (CCR2) inhibitor INCB3344 (HY-50674, MCE, USA) was delivered intraperitoneally at 30 mg/kg/day to VMC mice, starting on Day 1 and continuing until Day 14 post-CVB3 infection.

Techniques: Flow Cytometry, Control, Infection

Journal: Inflammation

Article Title: Activation of β2-Adrenergic Receptor Alleviates Viral Myocarditis by Regulating Energy Metabolism in Monocyte-derived Macrophages via the AMPK Pathway

doi: 10.1007/s10753-025-02382-6

Figure Lengend Snippet: Effects of activating β2-AR on the polarization and function of cardiac CCR2 + macrophages and CCR2 − macrophages. A - B Representative flow cytometry images (A) and statistical analysis (B) of CCR2 + iNOS + M1 macrophages among CD45 + CD64 + F4/80 + CCR2 + macrophages in the control, CVB3 + vehicle, and CVB3 + formoterol groups at 1 week and 2 weeks post-infection ( n = 6/group). C - F Representative flow cytometry images and statistical analysis of TNF-α (C-D) and CCL2 (E–F) secreted by CD45 + CD64 + F4/80 + CCR2 + macrophages in the control, CVB3 + vehicle, and CVB3 + formoterol groups at 1 week and 2 weeks post-infection ( n = 6/group). G - H Representative flow cytometry images (G) and statistical analysis (H) of CCR2 + CD206 + M2 macrophages among CD45 + CD64 + F4/80 + CCR2 + macrophages in the control, CVB3 + vehicle, and CVB3 + formoterol groups at 1 week and 2 weeks post-infection ( n = 6/group). I - J Representative flow cytometry images (I) and statistical analysis (J) of IL-10 secretion by CD45 + CD64 + F4/80 + CCR2 + macrophages in the control, CVB3 + vehicle, and CVB3 + formoterol groups at 1 week and 2 weeks post-infection ( n = 6/group). K - L Representative flow cytometry images (K) and statistical analysis (L) of CCR2 − iNOS + M1 macrophages among CD45 + CD64 + F4/80 + CCR2 − macrophages in the control, CVB3 + vehicle, and CVB3 + formoterol groups at 1 week and 2 weeks post-infection ( n = 6/group). M - P Representative flow cytometry images and statistical analysis of TNF-α (M–N) and CCL2 (O-P) secretion by CD45 + CD64 + F4/80 + CCR2 − macrophages in the control, CVB3 + vehicle, and CVB3 + formoterol groups at 1 week and 2 weeks post-infection ( n = 6/group). Q - R Representative flow cytometry images (Q) and statistical analysis (R) of CCR2 − CD206 + M2 macrophages among CD45 + CD64 + F4/80 + CCR2 − macrophages in the control, CVB3 + vehicle, and CVB3 + formoterol groups at 1 week and 2 weeks post-infection ( n = 6/group). S - T Representative flow cytometry images (S) and statistical analysis (T) of IL-10 secretion by CD45 + CD64 + F4/80 + CCR2 − macrophages in the control, CVB3 + vehicle, and CVB3 + formoterol groups at 1 week and 2 weeks post-infection ( n = 6/group). The data are presented as the means ± SEMs. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns = not significant. Abbreviations: CCL2, C–C motif chemokine ligand 2; CCR2, C–C chemokine receptor type 2; CVB3, coxsackievirus B3; IL-10, interleukin 10; iNOS, inducible nitric oxide synthase; SEM, standard error of the mean; TNF-α, tumor necrosis factor-α; β2-adrenergic receptor, β2-AR

Article Snippet: Following established protocols [ ], the C–C motif chemokine receptor 2 (CCR2) inhibitor INCB3344 (HY-50674, MCE, USA) was delivered intraperitoneally at 30 mg/kg/day to VMC mice, starting on Day 1 and continuing until Day 14 post-CVB3 infection.

Techniques: Flow Cytometry, Control, Infection

Journal: Inflammation

Article Title: Activation of β2-Adrenergic Receptor Alleviates Viral Myocarditis by Regulating Energy Metabolism in Monocyte-derived Macrophages via the AMPK Pathway

doi: 10.1007/s10753-025-02382-6

Figure Lengend Snippet: Activating β2-AR attenuates inflammation and fibrosis in VMC via CCR2 + macrophages. A Schematic illustrating macrophage depletion using clodronate liposomes (Lipo-Clod). The mice received intraperitoneal injections of 200 µl clodronate liposomes or PBS-containing liposomes (Lipo-PBS) 2 days before CVB3 infection, followed by injections on days 5 and 10 post-infection. B - C Representative flow cytometry images (B) and statistical analysis (C) of CD11b + Ly6G − monocytes among splenic CD45 + cells in the Vehicle and Lipo-Clod groups at 1 week and 2 weeks post-CVB3 infection ( n = 6/group). D Schematic showing CCR2 + monocyte/macrophage inhibition via INCB3344 which was administered from day 1 to day 7 or day 14 post-CVB3 infection. E – F Representative flow cytometry images (E) and statistical analysis (F) of Ly6C high monocytes among blood CD45 + CD11b + Ly6G − cells in the Vehicle and INCB3344 groups at 1 week and 2 weeks post-CVB3 infection ( n = 6/group). G - H Representative H&E staining (G) and Masson’s trichrome staining (H) images of the myocardium from the Vehicle, Lipo-Clod, Formoterol, and Formoterol + Lipo-Clod groups at 1 week and 2 weeks post-CVB3 infection (original magnification, 200 × ; scale bars = 50 µm). I - J Statistical analysis of myocardial pathological scores (I) and cardiac collagen volume fractions (J) in each group at 1 week and 2 weeks post-CVB3 infection ( n = 6/group). K - L Statistical analysis of myocardial pathological scores (K) and cardiac collagen volume fractions (L) in the Vehicle, INCB3344, Formoterol, and Formoterol + INCB3344 groups 1 week and 2 weeks post-CVB3 infection ( n = 6/group). M - N Statistical analysis of myocardial pathological scores (M) and cardiac collagen volume fractions (N) in each group at 1 week and 2 weeks post-CVB3 infection ( n = 6/group). The data are presented as the means ± SEMs. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns = not significant. Abbreviations: CCR2, C-C chemokine receptor type 2; CVB3, coxsackievirus B3; VMC, viral myocarditis; SEM, standard error of the mean; ns, not statistically significant; β2-adrenergic receptor, β2-AR

Article Snippet: Following established protocols [ ], the C–C motif chemokine receptor 2 (CCR2) inhibitor INCB3344 (HY-50674, MCE, USA) was delivered intraperitoneally at 30 mg/kg/day to VMC mice, starting on Day 1 and continuing until Day 14 post-CVB3 infection.

Techniques: Liposomes, Infection, Flow Cytometry, Inhibition, Staining

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Liver Sinusoidal Endothelial Cells Promote Metabolic Dysfunction-associated Steatohepatitis Progression via Interleukin-1R1-mediated Chemokine Production Induced by Macrophage-derived Interleukin-1β

doi: 10.1016/j.jcmgh.2025.101698

Figure Lengend Snippet: Spatial transcriptomic analysis reveals a dominant IL1B–IL1R1 axis between macrophages and LSECs, with increased chemokine expression in periportal ECs accompanied by increased macrophage proximity. ( A ) Workflow for the analysis of liver tissues from a patient with MASH (CosMx_MASH_sample_1) using CosMx. ( B ) Heatmap of the directional spatial proximity score across source-target cell type pairs. ( C ) Top 20 LR pairs enriched in macrophage → endothelial cell/LSEC interactions based on spatial LR scores. ( D ) Heatmaps of spatially weighted LR scores. From left to right: IL1B–IL1R1, CXCL10–CXCR3, and CCL2–CCR2. Higher values indicate the preferential enrichment of source→target interactions. ( E ) ( Left panel ) Positional relationship between each EC and cholangiocyte in the FOV. ( Right panel ) Comparison of the shortest distance from each EC to the nearest cholangiocyte. ( F ) ( Left panel ) Positional relationship between ECs and LSECs and macrophages in the FOV. The white circles represent a 20-μm radius from each EC. ( Right panel ) Comparison of the average number of macrophages within 20 μm of each EC.

Article Snippet: , Ccr2 , Mm99999051_gH , , , .

Techniques: Expressing, Comparison