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antibody against ccr7  (R&D Systems)


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    Structured Review

    R&D Systems antibody against ccr7
    <t>CCR7</t> + (c4) MoMFs drive MASLD progression through enhanced antigen presentation and are recruited via the CCL19/CCL21 axis. (A) Feature plots displaying the expression of H2-Eb2 , Ccr7 , Cd209a , and Ciita . (B) Flow cytometry analysis showing CCR7 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (C) Correlation between the proportion of CCR7 + MoMFs and the NAFLD activity score in the combined cohort of NCD-fed and MCD-fed mice. (D, E) Dot plots show the enriched GO (D) and KEGG (E) terms of DEGs in CCR7 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. (F) Dot plots show the average expression levels of selected DEGs. Dot size denotes the percentage of expressed cells, and the color bar denotes scaled average expression. (G) Flow cytometry analysis showing MHC-II expression levels in CCR7 + MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (H) Percentage of CCR7 + cells among total intrahepatic MoMFs in mice. (I, J) Plasma ALT and AST levels in each group. (K–M) Representative images (K) and quantification of H&E staining (L) and Sirius red staining (M) in liver paraffin sections. Scale bars, 200 μm. (N) Relative mRNA levels of Ccl19 (left) and Ccl21 (right) in liver tissues of NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice. (O) Feature plots displaying the expression of Top2a , Mki67 , and Stmn1 . (P) Flow cytometry analysis showing Ki67 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (Q) Dot plots show the enriched GO terms of DEGs in Ki67 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. (R) Feature plots displaying the expression of Cd3d , Gzma , and Gzmb . (S) Flow cytometry analysis showing CD3 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (T) Dot plots show the enriched GO terms of DEGs in CD3 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. Abbreviations: ALT, alanine transaminase; AST, aspartate aminotransferase; CDHFD, choline-deficient high-fat diet; HFD, high-fat diet; H&E, Hematoxylin and Eosin; MASLD, metabolic dysfunction–associated steatotic liver disease; MCD, methionine/choline-deficient diet; MoMFs, monocyte-derived macrophages; NAFLD, nonalcoholic fatty liver disease; NCD, normal control diet; UMAP, uniform manifold approximation and projection.
    Antibody Against Ccr7, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 126 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Single-cell profiling reveals hepatic monocyte-derived macrophages heterogeneity during steatotic liver disease"

    Article Title: Single-cell profiling reveals hepatic monocyte-derived macrophages heterogeneity during steatotic liver disease

    Journal: Hepatology Communications

    doi: 10.1097/HC9.0000000000000928

    CCR7 + (c4) MoMFs drive MASLD progression through enhanced antigen presentation and are recruited via the CCL19/CCL21 axis. (A) Feature plots displaying the expression of H2-Eb2 , Ccr7 , Cd209a , and Ciita . (B) Flow cytometry analysis showing CCR7 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (C) Correlation between the proportion of CCR7 + MoMFs and the NAFLD activity score in the combined cohort of NCD-fed and MCD-fed mice. (D, E) Dot plots show the enriched GO (D) and KEGG (E) terms of DEGs in CCR7 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. (F) Dot plots show the average expression levels of selected DEGs. Dot size denotes the percentage of expressed cells, and the color bar denotes scaled average expression. (G) Flow cytometry analysis showing MHC-II expression levels in CCR7 + MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (H) Percentage of CCR7 + cells among total intrahepatic MoMFs in mice. (I, J) Plasma ALT and AST levels in each group. (K–M) Representative images (K) and quantification of H&E staining (L) and Sirius red staining (M) in liver paraffin sections. Scale bars, 200 μm. (N) Relative mRNA levels of Ccl19 (left) and Ccl21 (right) in liver tissues of NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice. (O) Feature plots displaying the expression of Top2a , Mki67 , and Stmn1 . (P) Flow cytometry analysis showing Ki67 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (Q) Dot plots show the enriched GO terms of DEGs in Ki67 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. (R) Feature plots displaying the expression of Cd3d , Gzma , and Gzmb . (S) Flow cytometry analysis showing CD3 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (T) Dot plots show the enriched GO terms of DEGs in CD3 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. Abbreviations: ALT, alanine transaminase; AST, aspartate aminotransferase; CDHFD, choline-deficient high-fat diet; HFD, high-fat diet; H&E, Hematoxylin and Eosin; MASLD, metabolic dysfunction–associated steatotic liver disease; MCD, methionine/choline-deficient diet; MoMFs, monocyte-derived macrophages; NAFLD, nonalcoholic fatty liver disease; NCD, normal control diet; UMAP, uniform manifold approximation and projection.
    Figure Legend Snippet: CCR7 + (c4) MoMFs drive MASLD progression through enhanced antigen presentation and are recruited via the CCL19/CCL21 axis. (A) Feature plots displaying the expression of H2-Eb2 , Ccr7 , Cd209a , and Ciita . (B) Flow cytometry analysis showing CCR7 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (C) Correlation between the proportion of CCR7 + MoMFs and the NAFLD activity score in the combined cohort of NCD-fed and MCD-fed mice. (D, E) Dot plots show the enriched GO (D) and KEGG (E) terms of DEGs in CCR7 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. (F) Dot plots show the average expression levels of selected DEGs. Dot size denotes the percentage of expressed cells, and the color bar denotes scaled average expression. (G) Flow cytometry analysis showing MHC-II expression levels in CCR7 + MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (H) Percentage of CCR7 + cells among total intrahepatic MoMFs in mice. (I, J) Plasma ALT and AST levels in each group. (K–M) Representative images (K) and quantification of H&E staining (L) and Sirius red staining (M) in liver paraffin sections. Scale bars, 200 μm. (N) Relative mRNA levels of Ccl19 (left) and Ccl21 (right) in liver tissues of NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice. (O) Feature plots displaying the expression of Top2a , Mki67 , and Stmn1 . (P) Flow cytometry analysis showing Ki67 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (Q) Dot plots show the enriched GO terms of DEGs in Ki67 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. (R) Feature plots displaying the expression of Cd3d , Gzma , and Gzmb . (S) Flow cytometry analysis showing CD3 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (T) Dot plots show the enriched GO terms of DEGs in CD3 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. Abbreviations: ALT, alanine transaminase; AST, aspartate aminotransferase; CDHFD, choline-deficient high-fat diet; HFD, high-fat diet; H&E, Hematoxylin and Eosin; MASLD, metabolic dysfunction–associated steatotic liver disease; MCD, methionine/choline-deficient diet; MoMFs, monocyte-derived macrophages; NAFLD, nonalcoholic fatty liver disease; NCD, normal control diet; UMAP, uniform manifold approximation and projection.

    Techniques Used: Immunopeptidomics, Expressing, Flow Cytometry, Activity Assay, Clinical Proteomics, Staining, Derivative Assay, Control

    The basal cluster 1 (c1) is depleted during MASLD progression and acquires CD14 + and CCR7 + phenotypes under inflammatory stimulation in vitro. (A) Feature plots displaying the expression of Uba52 , Rpl27 , Rpl15 , and Ifi27l2a . (B) The typical flow cytometry gating strategy and statistical analysis of cluster 1 relative to total MoMFs in NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice. (C) Correlation between the proportion of cluster 1 and the NAFLD activity score in the combined cohort of NCD-fed and MCD-fed mice. (D) UMAP plots displaying the major cellular subsets identified in the NCD and MCD groups. (E) Trajectory analysis of clusters 0, 1, 4, and 5 using Monocle 2. (F) Heatmap displaying gene expression dynamics across pseudotime for selected marker genes in clusters 0, 1, 4, and 5. (G–I) Flow cytometry analysis of CD14 (G), CCR7 (H), and CCR3 (I) expression in cluster 1 cells isolated from NCD-fed mouse liver, with or without LPS stimulation. (J) Dot plots depicting the expression of genes in liver macrophages derived from healthy and MASLD samples ( GSE212837 ). Abbreviations: CDHFD, choline-deficient high-fat diet; HFD, high-fat diet; LPS, lipopolysaccharide; MASLD, metabolic dysfunction–associated steatotic liver disease; MCD, methionine/choline-deficient diet; MoMFs, monocyte-derived macrophages; NCD, normal control diet; UMAP, uniform manifold approximation and projection.
    Figure Legend Snippet: The basal cluster 1 (c1) is depleted during MASLD progression and acquires CD14 + and CCR7 + phenotypes under inflammatory stimulation in vitro. (A) Feature plots displaying the expression of Uba52 , Rpl27 , Rpl15 , and Ifi27l2a . (B) The typical flow cytometry gating strategy and statistical analysis of cluster 1 relative to total MoMFs in NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice. (C) Correlation between the proportion of cluster 1 and the NAFLD activity score in the combined cohort of NCD-fed and MCD-fed mice. (D) UMAP plots displaying the major cellular subsets identified in the NCD and MCD groups. (E) Trajectory analysis of clusters 0, 1, 4, and 5 using Monocle 2. (F) Heatmap displaying gene expression dynamics across pseudotime for selected marker genes in clusters 0, 1, 4, and 5. (G–I) Flow cytometry analysis of CD14 (G), CCR7 (H), and CCR3 (I) expression in cluster 1 cells isolated from NCD-fed mouse liver, with or without LPS stimulation. (J) Dot plots depicting the expression of genes in liver macrophages derived from healthy and MASLD samples ( GSE212837 ). Abbreviations: CDHFD, choline-deficient high-fat diet; HFD, high-fat diet; LPS, lipopolysaccharide; MASLD, metabolic dysfunction–associated steatotic liver disease; MCD, methionine/choline-deficient diet; MoMFs, monocyte-derived macrophages; NCD, normal control diet; UMAP, uniform manifold approximation and projection.

    Techniques Used: In Vitro, Expressing, Flow Cytometry, Activity Assay, Gene Expression, Marker, Isolation, Derivative Assay, Control

    Heterogeneity and dynamics of intrahepatic MoMFs during MASLD progression. This scRNA-seq analysis delineated 7 distinct MoMFs clusters (c0–c6) in the liver with dynamic changes during steatohepatitis progression. CD14 + (c0) MoMFs, the predominant CCR2 + population, exhibited multifaceted functions beyond inflammation, including roles in fibrosis, lipid uptake, and antigen presentation. The basal cluster (c1) is depleted during MASLD progression and acquires CD14 + and CCR7⁺ phenotypes under inflammatory stimulation in vitro. Lipid overload drives mitochondrial dysfunction and apoptosis in CD206 + (c2) MoMFs in MASLD. CCR3 + (c3) MoMFs drive MASLD progression through lipid accumulation, oxidative stress, and CCL5-mediated recruitment. CCL19/CCL21-recruited CCR7 + (c4) MoMFs exacerbate MASLD pathogenesis through enhanced antigen presentation. In addition, Ki67 + (c5) proliferative and CD3 + (c6) TCR-related MoMFs maintained stable proportions but exhibited functional alterations during MASLD. Abbreviations: MASLD, metabolic dysfunction–associated steatotic liver disease; MoMFs, monocyte-derived macrophages; scRNA-seq, single-cell RNA sequencing.
    Figure Legend Snippet: Heterogeneity and dynamics of intrahepatic MoMFs during MASLD progression. This scRNA-seq analysis delineated 7 distinct MoMFs clusters (c0–c6) in the liver with dynamic changes during steatohepatitis progression. CD14 + (c0) MoMFs, the predominant CCR2 + population, exhibited multifaceted functions beyond inflammation, including roles in fibrosis, lipid uptake, and antigen presentation. The basal cluster (c1) is depleted during MASLD progression and acquires CD14 + and CCR7⁺ phenotypes under inflammatory stimulation in vitro. Lipid overload drives mitochondrial dysfunction and apoptosis in CD206 + (c2) MoMFs in MASLD. CCR3 + (c3) MoMFs drive MASLD progression through lipid accumulation, oxidative stress, and CCL5-mediated recruitment. CCL19/CCL21-recruited CCR7 + (c4) MoMFs exacerbate MASLD pathogenesis through enhanced antigen presentation. In addition, Ki67 + (c5) proliferative and CD3 + (c6) TCR-related MoMFs maintained stable proportions but exhibited functional alterations during MASLD. Abbreviations: MASLD, metabolic dysfunction–associated steatotic liver disease; MoMFs, monocyte-derived macrophages; scRNA-seq, single-cell RNA sequencing.

    Techniques Used: Immunopeptidomics, In Vitro, Functional Assay, Derivative Assay, Single Cell, RNA Sequencing



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    CCR7 + (c4) MoMFs drive MASLD progression through enhanced antigen presentation and are recruited via the CCL19/CCL21 axis. (A) Feature plots displaying the expression of H2-Eb2 , Ccr7 , Cd209a , and Ciita . (B) Flow cytometry analysis showing CCR7 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (C) Correlation between the proportion of CCR7 + MoMFs and the NAFLD activity score in the combined cohort of NCD-fed and MCD-fed mice. (D, E) Dot plots show the enriched GO (D) and KEGG (E) terms of DEGs in CCR7 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. (F) Dot plots show the average expression levels of selected DEGs. Dot size denotes the percentage of expressed cells, and the color bar denotes scaled average expression. (G) Flow cytometry analysis showing MHC-II expression levels in CCR7 + MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (H) Percentage of CCR7 + cells among total intrahepatic MoMFs in mice. (I, J) Plasma ALT and AST levels in each group. (K–M) Representative images (K) and quantification of H&E staining (L) and Sirius red staining (M) in liver paraffin sections. Scale bars, 200 μm. (N) Relative mRNA levels of Ccl19 (left) and Ccl21 (right) in liver tissues of NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice. (O) Feature plots displaying the expression of Top2a , Mki67 , and Stmn1 . (P) Flow cytometry analysis showing Ki67 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (Q) Dot plots show the enriched GO terms of DEGs in Ki67 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. (R) Feature plots displaying the expression of Cd3d , Gzma , and Gzmb . (S) Flow cytometry analysis showing CD3 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (T) Dot plots show the enriched GO terms of DEGs in CD3 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. Abbreviations: ALT, alanine transaminase; AST, aspartate aminotransferase; CDHFD, choline-deficient high-fat diet; HFD, high-fat diet; H&E, Hematoxylin and Eosin; MASLD, metabolic dysfunction–associated steatotic liver disease; MCD, methionine/choline-deficient diet; MoMFs, monocyte-derived macrophages; NAFLD, nonalcoholic fatty liver disease; NCD, normal control diet; UMAP, uniform manifold approximation and projection.

    Journal: Hepatology Communications

    Article Title: Single-cell profiling reveals hepatic monocyte-derived macrophages heterogeneity during steatotic liver disease

    doi: 10.1097/HC9.0000000000000928

    Figure Lengend Snippet: CCR7 + (c4) MoMFs drive MASLD progression through enhanced antigen presentation and are recruited via the CCL19/CCL21 axis. (A) Feature plots displaying the expression of H2-Eb2 , Ccr7 , Cd209a , and Ciita . (B) Flow cytometry analysis showing CCR7 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (C) Correlation between the proportion of CCR7 + MoMFs and the NAFLD activity score in the combined cohort of NCD-fed and MCD-fed mice. (D, E) Dot plots show the enriched GO (D) and KEGG (E) terms of DEGs in CCR7 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. (F) Dot plots show the average expression levels of selected DEGs. Dot size denotes the percentage of expressed cells, and the color bar denotes scaled average expression. (G) Flow cytometry analysis showing MHC-II expression levels in CCR7 + MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (H) Percentage of CCR7 + cells among total intrahepatic MoMFs in mice. (I, J) Plasma ALT and AST levels in each group. (K–M) Representative images (K) and quantification of H&E staining (L) and Sirius red staining (M) in liver paraffin sections. Scale bars, 200 μm. (N) Relative mRNA levels of Ccl19 (left) and Ccl21 (right) in liver tissues of NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice. (O) Feature plots displaying the expression of Top2a , Mki67 , and Stmn1 . (P) Flow cytometry analysis showing Ki67 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (Q) Dot plots show the enriched GO terms of DEGs in Ki67 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. (R) Feature plots displaying the expression of Cd3d , Gzma , and Gzmb . (S) Flow cytometry analysis showing CD3 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (T) Dot plots show the enriched GO terms of DEGs in CD3 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. Abbreviations: ALT, alanine transaminase; AST, aspartate aminotransferase; CDHFD, choline-deficient high-fat diet; HFD, high-fat diet; H&E, Hematoxylin and Eosin; MASLD, metabolic dysfunction–associated steatotic liver disease; MCD, methionine/choline-deficient diet; MoMFs, monocyte-derived macrophages; NAFLD, nonalcoholic fatty liver disease; NCD, normal control diet; UMAP, uniform manifold approximation and projection.

    Article Snippet: To further determine the functional contribution of CCR7 + MoMFs, we administered a neutralizing antibody against CCR7 (MAB3477, R&D Systems) or an IgG2A isotype (clone 4B12, R&D Systems) to MCD-fed mice.

    Techniques: Immunopeptidomics, Expressing, Flow Cytometry, Activity Assay, Clinical Proteomics, Staining, Derivative Assay, Control

    The basal cluster 1 (c1) is depleted during MASLD progression and acquires CD14 + and CCR7 + phenotypes under inflammatory stimulation in vitro. (A) Feature plots displaying the expression of Uba52 , Rpl27 , Rpl15 , and Ifi27l2a . (B) The typical flow cytometry gating strategy and statistical analysis of cluster 1 relative to total MoMFs in NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice. (C) Correlation between the proportion of cluster 1 and the NAFLD activity score in the combined cohort of NCD-fed and MCD-fed mice. (D) UMAP plots displaying the major cellular subsets identified in the NCD and MCD groups. (E) Trajectory analysis of clusters 0, 1, 4, and 5 using Monocle 2. (F) Heatmap displaying gene expression dynamics across pseudotime for selected marker genes in clusters 0, 1, 4, and 5. (G–I) Flow cytometry analysis of CD14 (G), CCR7 (H), and CCR3 (I) expression in cluster 1 cells isolated from NCD-fed mouse liver, with or without LPS stimulation. (J) Dot plots depicting the expression of genes in liver macrophages derived from healthy and MASLD samples ( GSE212837 ). Abbreviations: CDHFD, choline-deficient high-fat diet; HFD, high-fat diet; LPS, lipopolysaccharide; MASLD, metabolic dysfunction–associated steatotic liver disease; MCD, methionine/choline-deficient diet; MoMFs, monocyte-derived macrophages; NCD, normal control diet; UMAP, uniform manifold approximation and projection.

    Journal: Hepatology Communications

    Article Title: Single-cell profiling reveals hepatic monocyte-derived macrophages heterogeneity during steatotic liver disease

    doi: 10.1097/HC9.0000000000000928

    Figure Lengend Snippet: The basal cluster 1 (c1) is depleted during MASLD progression and acquires CD14 + and CCR7 + phenotypes under inflammatory stimulation in vitro. (A) Feature plots displaying the expression of Uba52 , Rpl27 , Rpl15 , and Ifi27l2a . (B) The typical flow cytometry gating strategy and statistical analysis of cluster 1 relative to total MoMFs in NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice. (C) Correlation between the proportion of cluster 1 and the NAFLD activity score in the combined cohort of NCD-fed and MCD-fed mice. (D) UMAP plots displaying the major cellular subsets identified in the NCD and MCD groups. (E) Trajectory analysis of clusters 0, 1, 4, and 5 using Monocle 2. (F) Heatmap displaying gene expression dynamics across pseudotime for selected marker genes in clusters 0, 1, 4, and 5. (G–I) Flow cytometry analysis of CD14 (G), CCR7 (H), and CCR3 (I) expression in cluster 1 cells isolated from NCD-fed mouse liver, with or without LPS stimulation. (J) Dot plots depicting the expression of genes in liver macrophages derived from healthy and MASLD samples ( GSE212837 ). Abbreviations: CDHFD, choline-deficient high-fat diet; HFD, high-fat diet; LPS, lipopolysaccharide; MASLD, metabolic dysfunction–associated steatotic liver disease; MCD, methionine/choline-deficient diet; MoMFs, monocyte-derived macrophages; NCD, normal control diet; UMAP, uniform manifold approximation and projection.

    Article Snippet: To further determine the functional contribution of CCR7 + MoMFs, we administered a neutralizing antibody against CCR7 (MAB3477, R&D Systems) or an IgG2A isotype (clone 4B12, R&D Systems) to MCD-fed mice.

    Techniques: In Vitro, Expressing, Flow Cytometry, Activity Assay, Gene Expression, Marker, Isolation, Derivative Assay, Control

    Heterogeneity and dynamics of intrahepatic MoMFs during MASLD progression. This scRNA-seq analysis delineated 7 distinct MoMFs clusters (c0–c6) in the liver with dynamic changes during steatohepatitis progression. CD14 + (c0) MoMFs, the predominant CCR2 + population, exhibited multifaceted functions beyond inflammation, including roles in fibrosis, lipid uptake, and antigen presentation. The basal cluster (c1) is depleted during MASLD progression and acquires CD14 + and CCR7⁺ phenotypes under inflammatory stimulation in vitro. Lipid overload drives mitochondrial dysfunction and apoptosis in CD206 + (c2) MoMFs in MASLD. CCR3 + (c3) MoMFs drive MASLD progression through lipid accumulation, oxidative stress, and CCL5-mediated recruitment. CCL19/CCL21-recruited CCR7 + (c4) MoMFs exacerbate MASLD pathogenesis through enhanced antigen presentation. In addition, Ki67 + (c5) proliferative and CD3 + (c6) TCR-related MoMFs maintained stable proportions but exhibited functional alterations during MASLD. Abbreviations: MASLD, metabolic dysfunction–associated steatotic liver disease; MoMFs, monocyte-derived macrophages; scRNA-seq, single-cell RNA sequencing.

    Journal: Hepatology Communications

    Article Title: Single-cell profiling reveals hepatic monocyte-derived macrophages heterogeneity during steatotic liver disease

    doi: 10.1097/HC9.0000000000000928

    Figure Lengend Snippet: Heterogeneity and dynamics of intrahepatic MoMFs during MASLD progression. This scRNA-seq analysis delineated 7 distinct MoMFs clusters (c0–c6) in the liver with dynamic changes during steatohepatitis progression. CD14 + (c0) MoMFs, the predominant CCR2 + population, exhibited multifaceted functions beyond inflammation, including roles in fibrosis, lipid uptake, and antigen presentation. The basal cluster (c1) is depleted during MASLD progression and acquires CD14 + and CCR7⁺ phenotypes under inflammatory stimulation in vitro. Lipid overload drives mitochondrial dysfunction and apoptosis in CD206 + (c2) MoMFs in MASLD. CCR3 + (c3) MoMFs drive MASLD progression through lipid accumulation, oxidative stress, and CCL5-mediated recruitment. CCL19/CCL21-recruited CCR7 + (c4) MoMFs exacerbate MASLD pathogenesis through enhanced antigen presentation. In addition, Ki67 + (c5) proliferative and CD3 + (c6) TCR-related MoMFs maintained stable proportions but exhibited functional alterations during MASLD. Abbreviations: MASLD, metabolic dysfunction–associated steatotic liver disease; MoMFs, monocyte-derived macrophages; scRNA-seq, single-cell RNA sequencing.

    Article Snippet: To further determine the functional contribution of CCR7 + MoMFs, we administered a neutralizing antibody against CCR7 (MAB3477, R&D Systems) or an IgG2A isotype (clone 4B12, R&D Systems) to MCD-fed mice.

    Techniques: Immunopeptidomics, In Vitro, Functional Assay, Derivative Assay, Single Cell, RNA Sequencing

    CCR7 + (c4) MoMFs drive MASLD progression through enhanced antigen presentation and are recruited via the CCL19/CCL21 axis. (A) Feature plots displaying the expression of H2-Eb2 , Ccr7 , Cd209a , and Ciita . (B) Flow cytometry analysis showing CCR7 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (C) Correlation between the proportion of CCR7 + MoMFs and the NAFLD activity score in the combined cohort of NCD-fed and MCD-fed mice. (D, E) Dot plots show the enriched GO (D) and KEGG (E) terms of DEGs in CCR7 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. (F) Dot plots show the average expression levels of selected DEGs. Dot size denotes the percentage of expressed cells, and the color bar denotes scaled average expression. (G) Flow cytometry analysis showing MHC-II expression levels in CCR7 + MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (H) Percentage of CCR7 + cells among total intrahepatic MoMFs in mice. (I, J) Plasma ALT and AST levels in each group. (K–M) Representative images (K) and quantification of H&E staining (L) and Sirius red staining (M) in liver paraffin sections. Scale bars, 200 μm. (N) Relative mRNA levels of Ccl19 (left) and Ccl21 (right) in liver tissues of NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice. (O) Feature plots displaying the expression of Top2a , Mki67 , and Stmn1 . (P) Flow cytometry analysis showing Ki67 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (Q) Dot plots show the enriched GO terms of DEGs in Ki67 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. (R) Feature plots displaying the expression of Cd3d , Gzma , and Gzmb . (S) Flow cytometry analysis showing CD3 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (T) Dot plots show the enriched GO terms of DEGs in CD3 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. Abbreviations: ALT, alanine transaminase; AST, aspartate aminotransferase; CDHFD, choline-deficient high-fat diet; HFD, high-fat diet; H&E, Hematoxylin and Eosin; MASLD, metabolic dysfunction–associated steatotic liver disease; MCD, methionine/choline-deficient diet; MoMFs, monocyte-derived macrophages; NAFLD, nonalcoholic fatty liver disease; NCD, normal control diet; UMAP, uniform manifold approximation and projection.

    Journal: Hepatology Communications

    Article Title: Single-cell profiling reveals hepatic monocyte-derived macrophages heterogeneity during steatotic liver disease

    doi: 10.1097/HC9.0000000000000928

    Figure Lengend Snippet: CCR7 + (c4) MoMFs drive MASLD progression through enhanced antigen presentation and are recruited via the CCL19/CCL21 axis. (A) Feature plots displaying the expression of H2-Eb2 , Ccr7 , Cd209a , and Ciita . (B) Flow cytometry analysis showing CCR7 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (C) Correlation between the proportion of CCR7 + MoMFs and the NAFLD activity score in the combined cohort of NCD-fed and MCD-fed mice. (D, E) Dot plots show the enriched GO (D) and KEGG (E) terms of DEGs in CCR7 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. (F) Dot plots show the average expression levels of selected DEGs. Dot size denotes the percentage of expressed cells, and the color bar denotes scaled average expression. (G) Flow cytometry analysis showing MHC-II expression levels in CCR7 + MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (H) Percentage of CCR7 + cells among total intrahepatic MoMFs in mice. (I, J) Plasma ALT and AST levels in each group. (K–M) Representative images (K) and quantification of H&E staining (L) and Sirius red staining (M) in liver paraffin sections. Scale bars, 200 μm. (N) Relative mRNA levels of Ccl19 (left) and Ccl21 (right) in liver tissues of NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice. (O) Feature plots displaying the expression of Top2a , Mki67 , and Stmn1 . (P) Flow cytometry analysis showing Ki67 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (Q) Dot plots show the enriched GO terms of DEGs in Ki67 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. (R) Feature plots displaying the expression of Cd3d , Gzma , and Gzmb . (S) Flow cytometry analysis showing CD3 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (T) Dot plots show the enriched GO terms of DEGs in CD3 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. Abbreviations: ALT, alanine transaminase; AST, aspartate aminotransferase; CDHFD, choline-deficient high-fat diet; HFD, high-fat diet; H&E, Hematoxylin and Eosin; MASLD, metabolic dysfunction–associated steatotic liver disease; MCD, methionine/choline-deficient diet; MoMFs, monocyte-derived macrophages; NAFLD, nonalcoholic fatty liver disease; NCD, normal control diet; UMAP, uniform manifold approximation and projection.

    Article Snippet: To further determine the functional contribution of CCR7 + MoMFs, we administered a neutralizing antibody against CCR7 (MAB3477, R&D Systems) or an IgG2A isotype (clone 4B12, R&D Systems) to MCD-fed mice.

    Techniques: Immunopeptidomics, Expressing, Flow Cytometry, Activity Assay, Clinical Proteomics, Staining, Derivative Assay, Control

    The basal cluster 1 (c1) is depleted during MASLD progression and acquires CD14 + and CCR7 + phenotypes under inflammatory stimulation in vitro. (A) Feature plots displaying the expression of Uba52 , Rpl27 , Rpl15 , and Ifi27l2a . (B) The typical flow cytometry gating strategy and statistical analysis of cluster 1 relative to total MoMFs in NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice. (C) Correlation between the proportion of cluster 1 and the NAFLD activity score in the combined cohort of NCD-fed and MCD-fed mice. (D) UMAP plots displaying the major cellular subsets identified in the NCD and MCD groups. (E) Trajectory analysis of clusters 0, 1, 4, and 5 using Monocle 2. (F) Heatmap displaying gene expression dynamics across pseudotime for selected marker genes in clusters 0, 1, 4, and 5. (G–I) Flow cytometry analysis of CD14 (G), CCR7 (H), and CCR3 (I) expression in cluster 1 cells isolated from NCD-fed mouse liver, with or without LPS stimulation. (J) Dot plots depicting the expression of genes in liver macrophages derived from healthy and MASLD samples ( GSE212837 ). Abbreviations: CDHFD, choline-deficient high-fat diet; HFD, high-fat diet; LPS, lipopolysaccharide; MASLD, metabolic dysfunction–associated steatotic liver disease; MCD, methionine/choline-deficient diet; MoMFs, monocyte-derived macrophages; NCD, normal control diet; UMAP, uniform manifold approximation and projection.

    Journal: Hepatology Communications

    Article Title: Single-cell profiling reveals hepatic monocyte-derived macrophages heterogeneity during steatotic liver disease

    doi: 10.1097/HC9.0000000000000928

    Figure Lengend Snippet: The basal cluster 1 (c1) is depleted during MASLD progression and acquires CD14 + and CCR7 + phenotypes under inflammatory stimulation in vitro. (A) Feature plots displaying the expression of Uba52 , Rpl27 , Rpl15 , and Ifi27l2a . (B) The typical flow cytometry gating strategy and statistical analysis of cluster 1 relative to total MoMFs in NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice. (C) Correlation between the proportion of cluster 1 and the NAFLD activity score in the combined cohort of NCD-fed and MCD-fed mice. (D) UMAP plots displaying the major cellular subsets identified in the NCD and MCD groups. (E) Trajectory analysis of clusters 0, 1, 4, and 5 using Monocle 2. (F) Heatmap displaying gene expression dynamics across pseudotime for selected marker genes in clusters 0, 1, 4, and 5. (G–I) Flow cytometry analysis of CD14 (G), CCR7 (H), and CCR3 (I) expression in cluster 1 cells isolated from NCD-fed mouse liver, with or without LPS stimulation. (J) Dot plots depicting the expression of genes in liver macrophages derived from healthy and MASLD samples ( GSE212837 ). Abbreviations: CDHFD, choline-deficient high-fat diet; HFD, high-fat diet; LPS, lipopolysaccharide; MASLD, metabolic dysfunction–associated steatotic liver disease; MCD, methionine/choline-deficient diet; MoMFs, monocyte-derived macrophages; NCD, normal control diet; UMAP, uniform manifold approximation and projection.

    Article Snippet: To further determine the functional contribution of CCR7 + MoMFs, we administered a neutralizing antibody against CCR7 (MAB3477, R&D Systems) or an IgG2A isotype (clone 4B12, R&D Systems) to MCD-fed mice.

    Techniques: In Vitro, Expressing, Flow Cytometry, Activity Assay, Gene Expression, Marker, Isolation, Derivative Assay, Control

    Heterogeneity and dynamics of intrahepatic MoMFs during MASLD progression. This scRNA-seq analysis delineated 7 distinct MoMFs clusters (c0–c6) in the liver with dynamic changes during steatohepatitis progression. CD14 + (c0) MoMFs, the predominant CCR2 + population, exhibited multifaceted functions beyond inflammation, including roles in fibrosis, lipid uptake, and antigen presentation. The basal cluster (c1) is depleted during MASLD progression and acquires CD14 + and CCR7⁺ phenotypes under inflammatory stimulation in vitro. Lipid overload drives mitochondrial dysfunction and apoptosis in CD206 + (c2) MoMFs in MASLD. CCR3 + (c3) MoMFs drive MASLD progression through lipid accumulation, oxidative stress, and CCL5-mediated recruitment. CCL19/CCL21-recruited CCR7 + (c4) MoMFs exacerbate MASLD pathogenesis through enhanced antigen presentation. In addition, Ki67 + (c5) proliferative and CD3 + (c6) TCR-related MoMFs maintained stable proportions but exhibited functional alterations during MASLD. Abbreviations: MASLD, metabolic dysfunction–associated steatotic liver disease; MoMFs, monocyte-derived macrophages; scRNA-seq, single-cell RNA sequencing.

    Journal: Hepatology Communications

    Article Title: Single-cell profiling reveals hepatic monocyte-derived macrophages heterogeneity during steatotic liver disease

    doi: 10.1097/HC9.0000000000000928

    Figure Lengend Snippet: Heterogeneity and dynamics of intrahepatic MoMFs during MASLD progression. This scRNA-seq analysis delineated 7 distinct MoMFs clusters (c0–c6) in the liver with dynamic changes during steatohepatitis progression. CD14 + (c0) MoMFs, the predominant CCR2 + population, exhibited multifaceted functions beyond inflammation, including roles in fibrosis, lipid uptake, and antigen presentation. The basal cluster (c1) is depleted during MASLD progression and acquires CD14 + and CCR7⁺ phenotypes under inflammatory stimulation in vitro. Lipid overload drives mitochondrial dysfunction and apoptosis in CD206 + (c2) MoMFs in MASLD. CCR3 + (c3) MoMFs drive MASLD progression through lipid accumulation, oxidative stress, and CCL5-mediated recruitment. CCL19/CCL21-recruited CCR7 + (c4) MoMFs exacerbate MASLD pathogenesis through enhanced antigen presentation. In addition, Ki67 + (c5) proliferative and CD3 + (c6) TCR-related MoMFs maintained stable proportions but exhibited functional alterations during MASLD. Abbreviations: MASLD, metabolic dysfunction–associated steatotic liver disease; MoMFs, monocyte-derived macrophages; scRNA-seq, single-cell RNA sequencing.

    Article Snippet: To further determine the functional contribution of CCR7 + MoMFs, we administered a neutralizing antibody against CCR7 (MAB3477, R&D Systems) or an IgG2A isotype (clone 4B12, R&D Systems) to MCD-fed mice.

    Techniques: Immunopeptidomics, In Vitro, Functional Assay, Derivative Assay, Single Cell, RNA Sequencing

    CCR7 + (c4) MoMFs drive MASLD progression through enhanced antigen presentation and are recruited via the CCL19/CCL21 axis. (A) Feature plots displaying the expression of H2-Eb2 , Ccr7 , Cd209a , and Ciita . (B) Flow cytometry analysis showing CCR7 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (C) Correlation between the proportion of CCR7 + MoMFs and the NAFLD activity score in the combined cohort of NCD-fed and MCD-fed mice. (D, E) Dot plots show the enriched GO (D) and KEGG (E) terms of DEGs in CCR7 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. (F) Dot plots show the average expression levels of selected DEGs. Dot size denotes the percentage of expressed cells, and the color bar denotes scaled average expression. (G) Flow cytometry analysis showing MHC-II expression levels in CCR7 + MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (H) Percentage of CCR7 + cells among total intrahepatic MoMFs in mice. (I, J) Plasma ALT and AST levels in each group. (K–M) Representative images (K) and quantification of H&E staining (L) and Sirius red staining (M) in liver paraffin sections. Scale bars, 200 μm. (N) Relative mRNA levels of Ccl19 (left) and Ccl21 (right) in liver tissues of NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice. (O) Feature plots displaying the expression of Top2a , Mki67 , and Stmn1 . (P) Flow cytometry analysis showing Ki67 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (Q) Dot plots show the enriched GO terms of DEGs in Ki67 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. (R) Feature plots displaying the expression of Cd3d , Gzma , and Gzmb . (S) Flow cytometry analysis showing CD3 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (T) Dot plots show the enriched GO terms of DEGs in CD3 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. Abbreviations: ALT, alanine transaminase; AST, aspartate aminotransferase; CDHFD, choline-deficient high-fat diet; HFD, high-fat diet; H&E, Hematoxylin and Eosin; MASLD, metabolic dysfunction–associated steatotic liver disease; MCD, methionine/choline-deficient diet; MoMFs, monocyte-derived macrophages; NAFLD, nonalcoholic fatty liver disease; NCD, normal control diet; UMAP, uniform manifold approximation and projection.

    Journal: Hepatology Communications

    Article Title: Single-cell profiling reveals hepatic monocyte-derived macrophages heterogeneity during steatotic liver disease

    doi: 10.1097/HC9.0000000000000928

    Figure Lengend Snippet: CCR7 + (c4) MoMFs drive MASLD progression through enhanced antigen presentation and are recruited via the CCL19/CCL21 axis. (A) Feature plots displaying the expression of H2-Eb2 , Ccr7 , Cd209a , and Ciita . (B) Flow cytometry analysis showing CCR7 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (C) Correlation between the proportion of CCR7 + MoMFs and the NAFLD activity score in the combined cohort of NCD-fed and MCD-fed mice. (D, E) Dot plots show the enriched GO (D) and KEGG (E) terms of DEGs in CCR7 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. (F) Dot plots show the average expression levels of selected DEGs. Dot size denotes the percentage of expressed cells, and the color bar denotes scaled average expression. (G) Flow cytometry analysis showing MHC-II expression levels in CCR7 + MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (H) Percentage of CCR7 + cells among total intrahepatic MoMFs in mice. (I, J) Plasma ALT and AST levels in each group. (K–M) Representative images (K) and quantification of H&E staining (L) and Sirius red staining (M) in liver paraffin sections. Scale bars, 200 μm. (N) Relative mRNA levels of Ccl19 (left) and Ccl21 (right) in liver tissues of NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice. (O) Feature plots displaying the expression of Top2a , Mki67 , and Stmn1 . (P) Flow cytometry analysis showing Ki67 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (Q) Dot plots show the enriched GO terms of DEGs in Ki67 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. (R) Feature plots displaying the expression of Cd3d , Gzma , and Gzmb . (S) Flow cytometry analysis showing CD3 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (T) Dot plots show the enriched GO terms of DEGs in CD3 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. Abbreviations: ALT, alanine transaminase; AST, aspartate aminotransferase; CDHFD, choline-deficient high-fat diet; HFD, high-fat diet; H&E, Hematoxylin and Eosin; MASLD, metabolic dysfunction–associated steatotic liver disease; MCD, methionine/choline-deficient diet; MoMFs, monocyte-derived macrophages; NAFLD, nonalcoholic fatty liver disease; NCD, normal control diet; UMAP, uniform manifold approximation and projection.

    Article Snippet: To target CCR7 + MoMFs, mice received twice-weekly intravenous (i.v.) injections of an anti-mouse CCR7 neutralizing antibody (MAB3477, R&D Systems) at 10 μg per mouse or an IgG2A isotype control (clone 4B12, R&D Systems)., To target CCR2 + MoMFs, mice were fed an MCD diet for 4 weeks.

    Techniques: Immunopeptidomics, Expressing, Flow Cytometry, Activity Assay, Clinical Proteomics, Staining, Derivative Assay, Control

    The basal cluster 1 (c1) is depleted during MASLD progression and acquires CD14 + and CCR7 + phenotypes under inflammatory stimulation in vitro. (A) Feature plots displaying the expression of Uba52 , Rpl27 , Rpl15 , and Ifi27l2a . (B) The typical flow cytometry gating strategy and statistical analysis of cluster 1 relative to total MoMFs in NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice. (C) Correlation between the proportion of cluster 1 and the NAFLD activity score in the combined cohort of NCD-fed and MCD-fed mice. (D) UMAP plots displaying the major cellular subsets identified in the NCD and MCD groups. (E) Trajectory analysis of clusters 0, 1, 4, and 5 using Monocle 2. (F) Heatmap displaying gene expression dynamics across pseudotime for selected marker genes in clusters 0, 1, 4, and 5. (G–I) Flow cytometry analysis of CD14 (G), CCR7 (H), and CCR3 (I) expression in cluster 1 cells isolated from NCD-fed mouse liver, with or without LPS stimulation. (J) Dot plots depicting the expression of genes in liver macrophages derived from healthy and MASLD samples ( GSE212837 ). Abbreviations: CDHFD, choline-deficient high-fat diet; HFD, high-fat diet; LPS, lipopolysaccharide; MASLD, metabolic dysfunction–associated steatotic liver disease; MCD, methionine/choline-deficient diet; MoMFs, monocyte-derived macrophages; NCD, normal control diet; UMAP, uniform manifold approximation and projection.

    Journal: Hepatology Communications

    Article Title: Single-cell profiling reveals hepatic monocyte-derived macrophages heterogeneity during steatotic liver disease

    doi: 10.1097/HC9.0000000000000928

    Figure Lengend Snippet: The basal cluster 1 (c1) is depleted during MASLD progression and acquires CD14 + and CCR7 + phenotypes under inflammatory stimulation in vitro. (A) Feature plots displaying the expression of Uba52 , Rpl27 , Rpl15 , and Ifi27l2a . (B) The typical flow cytometry gating strategy and statistical analysis of cluster 1 relative to total MoMFs in NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice. (C) Correlation between the proportion of cluster 1 and the NAFLD activity score in the combined cohort of NCD-fed and MCD-fed mice. (D) UMAP plots displaying the major cellular subsets identified in the NCD and MCD groups. (E) Trajectory analysis of clusters 0, 1, 4, and 5 using Monocle 2. (F) Heatmap displaying gene expression dynamics across pseudotime for selected marker genes in clusters 0, 1, 4, and 5. (G–I) Flow cytometry analysis of CD14 (G), CCR7 (H), and CCR3 (I) expression in cluster 1 cells isolated from NCD-fed mouse liver, with or without LPS stimulation. (J) Dot plots depicting the expression of genes in liver macrophages derived from healthy and MASLD samples ( GSE212837 ). Abbreviations: CDHFD, choline-deficient high-fat diet; HFD, high-fat diet; LPS, lipopolysaccharide; MASLD, metabolic dysfunction–associated steatotic liver disease; MCD, methionine/choline-deficient diet; MoMFs, monocyte-derived macrophages; NCD, normal control diet; UMAP, uniform manifold approximation and projection.

    Article Snippet: To target CCR7 + MoMFs, mice received twice-weekly intravenous (i.v.) injections of an anti-mouse CCR7 neutralizing antibody (MAB3477, R&D Systems) at 10 μg per mouse or an IgG2A isotype control (clone 4B12, R&D Systems)., To target CCR2 + MoMFs, mice were fed an MCD diet for 4 weeks.

    Techniques: In Vitro, Expressing, Flow Cytometry, Activity Assay, Gene Expression, Marker, Isolation, Derivative Assay, Control

    Heterogeneity and dynamics of intrahepatic MoMFs during MASLD progression. This scRNA-seq analysis delineated 7 distinct MoMFs clusters (c0–c6) in the liver with dynamic changes during steatohepatitis progression. CD14 + (c0) MoMFs, the predominant CCR2 + population, exhibited multifaceted functions beyond inflammation, including roles in fibrosis, lipid uptake, and antigen presentation. The basal cluster (c1) is depleted during MASLD progression and acquires CD14 + and CCR7⁺ phenotypes under inflammatory stimulation in vitro. Lipid overload drives mitochondrial dysfunction and apoptosis in CD206 + (c2) MoMFs in MASLD. CCR3 + (c3) MoMFs drive MASLD progression through lipid accumulation, oxidative stress, and CCL5-mediated recruitment. CCL19/CCL21-recruited CCR7 + (c4) MoMFs exacerbate MASLD pathogenesis through enhanced antigen presentation. In addition, Ki67 + (c5) proliferative and CD3 + (c6) TCR-related MoMFs maintained stable proportions but exhibited functional alterations during MASLD. Abbreviations: MASLD, metabolic dysfunction–associated steatotic liver disease; MoMFs, monocyte-derived macrophages; scRNA-seq, single-cell RNA sequencing.

    Journal: Hepatology Communications

    Article Title: Single-cell profiling reveals hepatic monocyte-derived macrophages heterogeneity during steatotic liver disease

    doi: 10.1097/HC9.0000000000000928

    Figure Lengend Snippet: Heterogeneity and dynamics of intrahepatic MoMFs during MASLD progression. This scRNA-seq analysis delineated 7 distinct MoMFs clusters (c0–c6) in the liver with dynamic changes during steatohepatitis progression. CD14 + (c0) MoMFs, the predominant CCR2 + population, exhibited multifaceted functions beyond inflammation, including roles in fibrosis, lipid uptake, and antigen presentation. The basal cluster (c1) is depleted during MASLD progression and acquires CD14 + and CCR7⁺ phenotypes under inflammatory stimulation in vitro. Lipid overload drives mitochondrial dysfunction and apoptosis in CD206 + (c2) MoMFs in MASLD. CCR3 + (c3) MoMFs drive MASLD progression through lipid accumulation, oxidative stress, and CCL5-mediated recruitment. CCL19/CCL21-recruited CCR7 + (c4) MoMFs exacerbate MASLD pathogenesis through enhanced antigen presentation. In addition, Ki67 + (c5) proliferative and CD3 + (c6) TCR-related MoMFs maintained stable proportions but exhibited functional alterations during MASLD. Abbreviations: MASLD, metabolic dysfunction–associated steatotic liver disease; MoMFs, monocyte-derived macrophages; scRNA-seq, single-cell RNA sequencing.

    Article Snippet: To target CCR7 + MoMFs, mice received twice-weekly intravenous (i.v.) injections of an anti-mouse CCR7 neutralizing antibody (MAB3477, R&D Systems) at 10 μg per mouse or an IgG2A isotype control (clone 4B12, R&D Systems)., To target CCR2 + MoMFs, mice were fed an MCD diet for 4 weeks.

    Techniques: Immunopeptidomics, In Vitro, Functional Assay, Derivative Assay, Single Cell, RNA Sequencing

    CCR7 protein expression in IBC and non-IBC cell lines. ( A ) Immunoblot analysis of protein lysates from IBC (A3250, SUM149, IBC-3, SUM190) and non-IBC cell lines (MDA-MB-468, MCF-7, TF7D, SK-Br-3) expressed high levels of CCR7 in vitro. ( B ) Ratio of CCR7 compared to β-tubulin demonstrated similar levels of overall CCR7 protein expression between IBC and non-IBC cell lines.

    Journal: Scientific Reports

    Article Title: CCR7 immune cell receptor expression in inflammatory breast cancer

    doi: 10.1038/s41598-026-43437-4

    Figure Lengend Snippet: CCR7 protein expression in IBC and non-IBC cell lines. ( A ) Immunoblot analysis of protein lysates from IBC (A3250, SUM149, IBC-3, SUM190) and non-IBC cell lines (MDA-MB-468, MCF-7, TF7D, SK-Br-3) expressed high levels of CCR7 in vitro. ( B ) Ratio of CCR7 compared to β-tubulin demonstrated similar levels of overall CCR7 protein expression between IBC and non-IBC cell lines.

    Article Snippet: Immunohistochemical (IHC) staining for CCR7 (R&D systems, catalog no. MAB197-100) was performed using Leica Bond RX autostainer.

    Techniques: Expressing, Western Blot, In Vitro

    CCR7 and CCL21 expression in IBC versus non-IBC patient samples. ( A – D ) CCR7 and CCL21 expression were significantly higher among IBC patients ( n = 137) compared to non-IBC patients ( n = 252), ( p = 0.0007 and 0.0018, respectively). ( B ) Among IBC patients, HER2-positive and basal subtypes had significantly higher levels of CCR7 compared to luminal subtypes ( p = 0.0002 and 0.0161, respectively) and ( C ) ER-negative status exhibited higher CCR7 expression compared to ER-positive status ( p = 0.0098). ( E ) Similarly, HER2-positive subtypes demonstrated higher CCL21 expression compared to luminal subtypes ( p = 0.0423 and 0.0366), while ( F ) ER status did not impact CCL21 expression. ( G ) CCR7 positively correlated with expression of its ligand, CCL21 ( r = 0.39, p < 0.0001) and ( H ) CCL21 was correlated with LYVE-1, a lymphangiogenic marker in breast cancer ( r = 0.24, p < 0.0001).

    Journal: Scientific Reports

    Article Title: CCR7 immune cell receptor expression in inflammatory breast cancer

    doi: 10.1038/s41598-026-43437-4

    Figure Lengend Snippet: CCR7 and CCL21 expression in IBC versus non-IBC patient samples. ( A – D ) CCR7 and CCL21 expression were significantly higher among IBC patients ( n = 137) compared to non-IBC patients ( n = 252), ( p = 0.0007 and 0.0018, respectively). ( B ) Among IBC patients, HER2-positive and basal subtypes had significantly higher levels of CCR7 compared to luminal subtypes ( p = 0.0002 and 0.0161, respectively) and ( C ) ER-negative status exhibited higher CCR7 expression compared to ER-positive status ( p = 0.0098). ( E ) Similarly, HER2-positive subtypes demonstrated higher CCL21 expression compared to luminal subtypes ( p = 0.0423 and 0.0366), while ( F ) ER status did not impact CCL21 expression. ( G ) CCR7 positively correlated with expression of its ligand, CCL21 ( r = 0.39, p < 0.0001) and ( H ) CCL21 was correlated with LYVE-1, a lymphangiogenic marker in breast cancer ( r = 0.24, p < 0.0001).

    Article Snippet: Immunohistochemical (IHC) staining for CCR7 (R&D systems, catalog no. MAB197-100) was performed using Leica Bond RX autostainer.

    Techniques: Expressing, Marker

    CCR7 TMA scoring scales. Representative TMA images of CCR7 scoring scales by intensity, ( A ) 1 + intensity, ( B ) 2 + intensity, ( C ) 3 + intensity. Note that the example for 1 + staining intensity is not from this sample cohort as no patients in this study had 1 + CCR7 intensity.

    Journal: Scientific Reports

    Article Title: CCR7 immune cell receptor expression in inflammatory breast cancer

    doi: 10.1038/s41598-026-43437-4

    Figure Lengend Snippet: CCR7 TMA scoring scales. Representative TMA images of CCR7 scoring scales by intensity, ( A ) 1 + intensity, ( B ) 2 + intensity, ( C ) 3 + intensity. Note that the example for 1 + staining intensity is not from this sample cohort as no patients in this study had 1 + CCR7 intensity.

    Article Snippet: Immunohistochemical (IHC) staining for CCR7 (R&D systems, catalog no. MAB197-100) was performed using Leica Bond RX autostainer.

    Techniques: Staining