Journal: Journal of experimental & clinical cancer research : CR

Article Title: Gastric cancer-derived exosomal let-7 g-5p mediated by SERPINE1 promotes macrophage M2 polarization and gastric cancer progression.

doi: 10.1186/s13046-024-03269-4

Figure Lengend Snippet: Fig. 2 High SERPINE1 expression in GC cells promotes macrophage M2 polarization. tSNE visualization of nine single-cell clusters partitioned by unsu pervised cluster analysis, SERPINE1 expression of each single-cell, and SERPINE1 expression abundance of different single-cell clusters in the GSE134520 (A–C) and GSE167297 (D–F) datasets. (G) Flow cytometry analysis of the proportion of CD68+CD206+ macrophages in a Transwell co-culture system, with MKN45 and AGS cells overexpressing (oe_SERPINE1) or silencing SERPINE1 (shRNA#3 or sh_SERPINE1#3) in the upper chamber, and THP1 cells treated with PMA in the lower chamber. (H) Immunofluorescence staining of xenograft tumor tissues. Comparison of the proportion of M1 or M2 macrophage infiltra tion. Green indicates F4/80. Red indicates iNOS or Arg1 expression

Article Snippet: THP-1 cells were differentiated into macrophages using 150 ng/mL phorbol 12-myristate 13-acetate (PMA, Sigma) for 24 h and subsequently co-cultured with cancer-derived exosomes or GC cells in 6-well plates with 0.4-μm membranes for 72 h. Harvested macrophages were converted into single-cell suspensions, stained with Elab Fluor 488 anti-human CD68 (Mouse, 1:20, ElabScience) and APC anti-human CD206 (Mouse, 1:20, ElabScience) antibodies, and analyzed for CD68+CD206+ populations by flow cytometry (Accuri C6, BD).

Techniques: Expressing, Flow Cytometry, Co-Culture Assay, shRNA, Immunofluorescence, Staining, Comparison

Journal: Journal of experimental & clinical cancer research : CR

Article Title: Gastric cancer-derived exosomal let-7 g-5p mediated by SERPINE1 promotes macrophage M2 polarization and gastric cancer progression.

doi: 10.1186/s13046-024-03269-4

Figure Lengend Snippet: Fig. 5 SERPINE1-mediated gastric cancer-derived exosomes facilitate the polarization of THP1 cells into M2 macrophages. (A) Schematic representation of the extraction and identification of exosomes and the induction of macrophage polarization. Transmission electron microscopy (B), nanoparticle tracking analysis (C), and western blotting (D) were used to identify the morphology, particle size, and markers of exosomes. (E) Confocal laser scanning microscopy detected Dil-labeled exosomes (red) internalized by DAPI-labeled macrophages (blue). (F–G) Immunofluorescence analysis of the proportion of CD206+ cells in THP1 cells treated with exosomes. (H–I) Flow cytometry analysis of the proportion of CD68+CD206+ cells in THP1 cells treated with exosomes. (J–K) qRT-PCR analysis of M1 markers (iNOS and TNF-α) and M2 markers (TGF-β, IL-10, and Arg-1) in THP1 cells treated with exosomes. (L–N) Transwell migration and invasion assays of GC cells (upper chamber) co-cultured with macrophages (lower chamber) ingesting exosomes

Article Snippet: THP-1 cells were differentiated into macrophages using 150 ng/mL phorbol 12-myristate 13-acetate (PMA, Sigma) for 24 h and subsequently co-cultured with cancer-derived exosomes or GC cells in 6-well plates with 0.4-μm membranes for 72 h. Harvested macrophages were converted into single-cell suspensions, stained with Elab Fluor 488 anti-human CD68 (Mouse, 1:20, ElabScience) and APC anti-human CD206 (Mouse, 1:20, ElabScience) antibodies, and analyzed for CD68+CD206+ populations by flow cytometry (Accuri C6, BD).

Techniques: Derivative Assay, Extraction, Transmission Assay, Electron Microscopy, Western Blot, Confocal Laser Scanning Microscopy, Labeling, Immunofluorescence, Flow Cytometry, Quantitative RT-PCR, Migration, Cell Culture

Journal: Mucosal immunology

Article Title: A CCL24-dependent Pathway Augments Eosinophilic Airway Inflammation in House Dust Mite-challenged Cd163 −/− Mice

doi: 10.1038/mi.2015.94

Figure Lengend Snippet: (A) Confocal immunofluorescence microscopy images of BALF cells from an asthmatic subject showing CD163 expression by CD68 + AMΦs. The scale bar indicates 5 μm. (B) Gating strategy for identification of human CD163 + alveolar macrophages in BALF. Cellular debris was excluded using a forward light scatter/side scatter plot and doublets were excluded using width parameter on FSC and SSC properties. CD45 + cells, that co-expressed CD14 and CD68, were identified as alveolar macrophages using side scatter and CD45 bivariate plots from which lymphocytes had been excluded. A microscopic image of sorted CD45 + /CD14 + /CD68 + /CD163 + cells shows a cellular population possessing typical cellular characteristics of alveolar macrophages. (C) MFI of cell surface CD163 expression by CD45 + /CD14 + /CD68 + AMΦs in BALF from normal individuals and asthmatic subjects (n = 7, P < 0.008, paired t test). (D) A representative histogram overlay comparing cell surface CD163 expression by AMΦs from a normal individual and an asthmatic subject.

Article Snippet: Human BALF cell cytospin slides were fixed in 4% paraformaldehyde, incubated in blocking buffer (Aurion Blocking Solution, Electron Microscopy Sciences, Hatfield, PA) and reacted overnight with mouse anti-human CD68 (1:50 dilution, R & D Systems, Minneapolis, MN) and rabbit anti-human CD163 antibodies (1:50 dilution, Enzo Life Sciences, Inc., NY) diluted in 0.1% Aurion BSA-c (Electron Microscopy Sciences).

Techniques: Immunofluorescence, Microscopy, Expressing

Journal: Cancers

Article Title: Crosstalk between Macrophages and Myxoid Liposarcoma Cells Increases Spreading and Invasiveness of Tumor Cells

doi: 10.3390/cancers13133298

Figure Lengend Snippet: Immunohistochemical characterization of immune infiltrate in MLPS tissues. Representative images of H&E and IHC staining of CD3+, CD4+, CD8+, CD68+, and CD163+ positive cells in FFPE sections from a low grade MLPS ( a ) from patient #21 and high grade. MLPS tissues ( b ) from patient #4, acquired at 200× magnification.

Article Snippet: Monocytes recovered from co-cultures with/without MLPS cells were analyzed by flow cytometry as described [ ], using PE-conjugated anti-CD68 REAfinity™ (Miltenyi Biotec #130-114-460) and APC-conjugated anti-CD163 REAfinity™ (Miltenyi Biotec, #130-112–129, Bergisch Gladbach, Germany) antibodies.

Techniques: Immunohistochemical staining, Immunohistochemistry

Journal: Cancers

Article Title: Crosstalk between Macrophages and Myxoid Liposarcoma Cells Increases Spreading and Invasiveness of Tumor Cells

doi: 10.3390/cancers13133298

Figure Lengend Snippet: Correlation between immune cell infiltration and microvessel density in 26 low grade versus 24 high grade MLPS tissues. Box plots, showing variation in the average count of CD3+, CD4+, CD8+, FOXP3+, CD68+, and CD163+ cells as well as CD31 positive microvessels according to low or high histologic grade. Dark horizontal lines indicate the medians. Circles indicate outliers.

Article Snippet: Monocytes recovered from co-cultures with/without MLPS cells were analyzed by flow cytometry as described [ ], using PE-conjugated anti-CD68 REAfinity™ (Miltenyi Biotec #130-114-460) and APC-conjugated anti-CD163 REAfinity™ (Miltenyi Biotec, #130-112–129, Bergisch Gladbach, Germany) antibodies.

Techniques:

Journal: Cancers

Article Title: Crosstalk between Macrophages and Myxoid Liposarcoma Cells Increases Spreading and Invasiveness of Tumor Cells

doi: 10.3390/cancers13133298

Figure Lengend Snippet: Pearson correlation between the average count of CD163+ macrophages and ( a ) CD3+ T lymphocytes, ( b ) CD4+ T helper lymphocytes, ( c ) CD8+ cytotoxic lymphocytes, ( d ) CD68+ macrophages, and ( e ) CD31+ microvessels in 50 MLPS tissue sections. Pearson correlation coefficients (r) are indicated.

Article Snippet: Monocytes recovered from co-cultures with/without MLPS cells were analyzed by flow cytometry as described [ ], using PE-conjugated anti-CD68 REAfinity™ (Miltenyi Biotec #130-114-460) and APC-conjugated anti-CD163 REAfinity™ (Miltenyi Biotec, #130-112–129, Bergisch Gladbach, Germany) antibodies.

Techniques:

Journal: Cancers

Article Title: Crosstalk between Macrophages and Myxoid Liposarcoma Cells Increases Spreading and Invasiveness of Tumor Cells

doi: 10.3390/cancers13133298

Figure Lengend Snippet: MLPS cells trigger M2-like polarization of monocytes in non-contact co-cultures. ( a , b ) Representative images of H&E staining of FFPE sections from #37 ( a ) and #47 ( b ) MLPS tissues acquired at 200× magnification. ( c , d ) Primary MLPS cells obtained by enzymatic digestion of 37 ( c ) and #47 ( d ) tumor tissues, visualized by phase contrast microscopy (left) and fluorescent microscopy after immunostaining with anti-vimentin and anti-cytokeratin Abs (right). Nuclei were stained blue with DAPI. Original magnifications: 200× (left) and 400× (right). ( e , f ) Human monocytes were co-cultured with #37 ( e ) and #47 ( f ) primary MLPS cells in an in vitro non-contact co-culture for 72 h and then analyzed for CD68 and CD163 expression by flow cytometry. ( e , f ) Percent variation of CD68 and CD163 on monocytes collected after non-contact co-culture, compared to control monocytes. ( g , h ) After co-cultures with #37 ( g ) and #47 ( h ) primary MLPS cells, CMs from monocytes were analyzed for the content of CC2, IL-10, and IL-12 by a dot plot assay. The pixel density of each spot was measured using NIH Image J 2.0 software developed by the US NIH, USA and positive control spots were used to normalize results between the membranes. The intensity of each spot was averaged over the duplicate spots and expressed as percentage of each cytokine or chemokine spontaneously secreted by control monocytes (monocyte CM), considered as 100% (dashed line). Data represent mean ± SD from three experiments performed in duplicate with * p < 0.05, ** p < 0.005.

Article Snippet: Monocytes recovered from co-cultures with/without MLPS cells were analyzed by flow cytometry as described [ ], using PE-conjugated anti-CD68 REAfinity™ (Miltenyi Biotec #130-114-460) and APC-conjugated anti-CD163 REAfinity™ (Miltenyi Biotec, #130-112–129, Bergisch Gladbach, Germany) antibodies.

Techniques: Staining, Microscopy, Immunostaining, Cell Culture, In Vitro, Co-Culture Assay, Expressing, Flow Cytometry, Software, Positive Control

Journal: JTCVS Open

Article Title: Evaluation of tissue-engineered human acellular vessels as a Blalock–Taussig–Thomas shunt in a juvenile primate model

doi: 10.1016/j.xjon.2023.05.018

Figure Lengend Snippet: Histologic evaluation of host immune response. Midgraft sections from 3.5 mm HAV mBTT shunt explants stained by IHC for CD20 (B-cell), CD3 (T-cell), CD68 (macrophage), and CD11b (monocyte) markers. NHPs with midgraft dilatations (NHP1 and NHP3 [J-O]) exhibited markedly greater levels of host immune infiltration within the mBTT shunt wall than NHPs with no dilatations (NHP2, NHP4, NHP5 [A-I]). Host immune cells in explants from NHP1 and NHP3 were predominantly localized around the regions of dilatation. H&E , Hematoxylin and Eosin; NHP , nonhuman primate; HAV , Human Acellular Vessel; mBTT , modified Blalock–Taussig–Thomas shunt; IHC , immunohistochemistry; DAPI , 4′,6-diamidino-2-phenylindole.

Article Snippet: Explanted tissue sections and tissue slides (American MasterTech) were immunostained for alpha-smooth muscle actin (α-SMA; Dako M0851), von Willebrand Factor (vWF; Abcam 179451), CD3 (Dako A0451), CD20 (Abcam ab9475), CD11b (Abcam 52478), and CD68 (R&D Systems MAB2040).

Techniques: Staining, Modification, Immunohistochemistry

Journal: Advanced Materials (Deerfield Beach, Fla.)

Article Title: A Pseudo‐ Mytilus Edulis Foot Protein‐Based Hydrogel Adhesive with Osteo‐Vascular‐Immune Coupling Effects for Osteoporotic Bone‐Implant Integration

doi: 10.1002/adma.202511840

Figure Lengend Snippet: Bioglue effectively modulates the immune microenvironment at the screw‒bone interface. A) Schematic diagram of the animal experimental design (created with BioRender.com). B) H&E staining of the screw‒bone interface tissue. C) Immunofluorescence staining of macrophage infiltration at the screw‐bone interface. D) IHC analysis of the screw‒bone interface. E) Senescence‐associated immunofluorescence staining of the screw‒bone interface tissue. F,G) Proportion of iNOS + and Arg‐1 + macrophages (iNOS + & CD68 + /Arg‐1 + & CD68 + ) in tissue fluorescent‐stained sections ( n = 5). The data are presented as the means ± SDs in the analysis figures, with * p < 0.05 and ** p < 0.01 indicating statistical significance.

Article Snippet: Sections were incubated with primary antibodies, iNOS (ABclonal, A3774), Arg‐1 (ABclonal, A25808), CD68 (Proteintech, 3A9A7), P21 (ABclonal, A19094), VEGF (ABclonal, A19094), IL‐10 (ABclonal, A2171), TNF‐α (ABclonal, A11534), at 4 °C overnight.

Techniques: Staining, Immunofluorescence

Journal: Cancer Research Communications

Article Title: Phase I First-in-Human Study of TRK-950, an IgG1 Antibody Specific to CAPRIN-1, in Patients with Advanced Solid Tumors

doi: 10.1158/2767-9764.CRC-25-0123

Figure Lengend Snippet: Graphs depicting the density of macrophage subsets pre- and posttreatment with TRK-950. Tumor specimens obtained with core-needle biopsies at the screening and C1D22 time points were subjected to multi-immunofluorescence to detect macrophage infiltrates. Quantitative analyses were performed on digital images of CD68 and CD163 staining. A, total CD68 + cells; ( B ) CD68 + /CD163 − subsets; ( C ) CD68 + /CD163 + subsets. Error bars indicate SD. C1D22, cycle 1 day 22; SCR, screening.

Article Snippet: After antigen retrieval (36 minutes at 95°C, pH 8.4), 4-μm-thick sections of tumor specimens obtained from core-needle biopsies at the screening and C1D22 time points were stained with mouse monoclonal anti-human CD68 (IgG2b, clone 298807, R&D Systems) and mouse monoclonal anti-human CD163 (IgG1, clone 10D6, Leica Biosystems) and then incubated with AF647 goat anti-mouse IgG2b and AF488 goat anti-mouse IgG1 (both from Invitrogen).

Techniques: Immunofluorescence, Staining

Journal: Frontiers in Immunology

Article Title: Increased expression of CD36 and CD163 in clear cell renal cell carcinoma suggests an association between lipid transport and an “M2-like” macrophage phenotype

doi: 10.3389/fimmu.2026.1773666

Figure Lengend Snippet: Expression of CD68 and CD163 in ccRCC samples. (A) CD68 was assessed using immunohistochemistry. (B) Expression of CD68 was assessed using the available TCGA data (KIRC dataset). Tumors were grouped by the pathological tumor size (pT). (C) CD163 was assessed using immunohistochemistry. (D) Expression of CD163 was assessed using the available TCGA data (KIRC dataset). Tumors were grouped by the pathological tumor size (pT). Student’s t-test for two groups and one-way ANOVA for four groups; *p<0.05, **p<0.01, ***p<0.001,****p<0.0001. (E) Multiplex immunofluorescence imaging of ccRCC tumor tissues (representative of three patients) depicting CK (cytokeratin), CD163 and CD68 expression. White bar represents 200 μm (left) and 10 μm (right). (F) UMAP depicting clusters of single-cell data showing the expression of CD68 and CD163. Cell type annotations were adopted from the original publication . (G) Fraction of CD68+ cells co-expressing CD163 in RCC tumor tissue macrophage population. Positivity in scRNA-seq for CD68 and CD163 was defined from raw UMI counts as ≥1 UMI per gene (F) . (H) Flow cytometric analysis of CD45 on cells from central and peripheral ccRCC tumor tissue and adjacent kidney. (I) As in (H) , analysis of CD163 on CD68+ cells from central and peripheral ccRCC tumor tissue and adjacent kidney. Values were normalized to kidney controls. Representative histogram with geometric mean fluorescence intensities on the right. One-way ANOVA with Dunnett post-test comparing to kidney. *p<0.05, **p<0.01, ***p<0.001. (J) Data from (I) , tumor periphery, plotted as individual patients, depicting the portion of CD163+ and CD163neg cells.

Article Snippet: Reagents and antibodies used included FcR Blocking Reagent (Cat# 130-059-901), CD36 PE (Cat# 130-110-877), CD147 APC (Cat# 130-124-295), CD8a PE (Cat# 130-117-201), CD45 PE (Cat# 130-113-118), CD68 PE (Cat# 130-128-345), CD163 PE (Cat# 130-127-908), and pan-Cytokeratin APC (Cat# 130-123-091), all from Miltenyi Biotec.

Techniques: Expressing, Immunohistochemistry, Multiplex Assay, Immunofluorescence, Imaging, Single Cell, Fluorescence

Journal: Frontiers in Immunology

Article Title: Increased expression of CD36 and CD163 in clear cell renal cell carcinoma suggests an association between lipid transport and an “M2-like” macrophage phenotype

doi: 10.3389/fimmu.2026.1773666

Figure Lengend Snippet: Correlations of CD36 and Oil Red O with immunological markers. Data was obtained as in Figures 1 , . Observer-based histological scores were used to calculate the correlations. As these scores are ordinal, individual data points may overlap in scatter plots. The number of overlapping values is indicated by numbers in parentheses. Correlation statistics were performed using Pearson´s correlation coefficients, and p-values were two-tailed. (A) Correlations of CD36 with CD68, CD163 and CD3. (B) Correlations of Oil Red O scores to CD68, CD163 and CD3. (C) Multiplex immunofluorescence imaging of ccRCC tumor tissues (representative of three patients) depicting CD68 and CD36 expression. (D) Flow cytometric analysis of CD36 on CD68+ cells from central and peripheral ccRCC tumor tissue and adjacent kidney. Values were normalized to kidney controls. Representative histogram with geometric mean fluorescence intensities on the right. One-way ANOVA with Dunnett post-test comparing to kidney, n.s. (E) As in (D) , correlation of CD36 and CD163 expression on CD68+ cells. (F) UMAP depicting macrophage cluster with expression of CD68, CD163 and CD36 . (G-I) Flow cytometric analysis of peripheral tumor tissues, correlating the expression of CD36 on CD45neg cells to (G) CD163 on CD68+ cells, (H) the CD68+ cell frequencies and (I) the frequencies of CD3+ CD8+ cells. Pearson´s correlation, two-tailed p-value. (J, K) Lipidomics performed on five ccRCC tumors with correlations of CD163 expression (IHC, area staining intensity in J; histological score in K) and the levels of triacylglycerol (TG).

Article Snippet: Reagents and antibodies used included FcR Blocking Reagent (Cat# 130-059-901), CD36 PE (Cat# 130-110-877), CD147 APC (Cat# 130-124-295), CD8a PE (Cat# 130-117-201), CD45 PE (Cat# 130-113-118), CD68 PE (Cat# 130-128-345), CD163 PE (Cat# 130-127-908), and pan-Cytokeratin APC (Cat# 130-123-091), all from Miltenyi Biotec.

Techniques: Two Tailed Test, Multiplex Assay, Immunofluorescence, Imaging, Expressing, Fluorescence, Staining

Journal: Frontiers in Immunology

Article Title: Increased expression of CD36 and CD163 in clear cell renal cell carcinoma suggests an association between lipid transport and an “M2-like” macrophage phenotype

doi: 10.3389/fimmu.2026.1773666

Figure Lengend Snippet: Expression of CD147 in ccRCC samples. (A) CD147 was assessed using immunohistochemistry. Student’s t-test, *p<0.05. (B) Expression of CD147 was assessed using the available TCGA data (KIRC dataset). (C) Correlations of CD147 expression to CD36 and CD163 using area staining intensity. (D) Correlations of CD147 expression to CD36 and CD163 using observer-based histological scores. As these scores are ordinal, individual data points may overlap in scatter plots. The number of overlapping values is indicated by numbers in parentheses. Correlation statistics were performed using Pearson´s correlation coefficients, and p-values were two-tailed. (E) Flow cytometric analysis of CD147 on CD45neg cells from central and peripheral ccRCC tumor tissue and adjacent kidney. Values were normalized to kidney controls. Representative histogram with geometric mean fluorescence intensities on the right. (F) As in (E) , correlations of CD147 expression on CD45neg cells to CD36 in tumor periphery and tumor center. (G) Multiplex immunofluorescence imaging of ccRCC tumor tissues (representative of three patients) depicting CD36 and CD147 expression. White bar represents 200 µm (left) and 10 µm (right). (H) UMAP depicting clusters of single-cell data from ccRCC patients showing the expression of CD147. Cell type annotations were adopted from the original publication . (I) . Flow cytometric analysis of CD147 on CD68+ cells from central and peripheral ccRCC tumor tissue and adjacent kidney. Values were normalized to kidney controls. Representative histogram with geometric mean fluorescence intensities on the right. (J) As in (I) , correlation of CD163 and CD147 expression on CD68+ cells. Pearson´s correlation, two-tailed p-value. (K) UMAP visualization of CD163 and BSG (CD147) expression on myeloid cells from ccRCC tumors, cell type annotations were adopted from the original publication .

Article Snippet: Reagents and antibodies used included FcR Blocking Reagent (Cat# 130-059-901), CD36 PE (Cat# 130-110-877), CD147 APC (Cat# 130-124-295), CD8a PE (Cat# 130-117-201), CD45 PE (Cat# 130-113-118), CD68 PE (Cat# 130-128-345), CD163 PE (Cat# 130-127-908), and pan-Cytokeratin APC (Cat# 130-123-091), all from Miltenyi Biotec.

Techniques: Expressing, Immunohistochemistry, Staining, Two Tailed Test, Fluorescence, Multiplex Assay, Immunofluorescence, Imaging, Single Cell

Journal: Frontiers in Immunology

Article Title: Increased expression of CD36 and CD163 in clear cell renal cell carcinoma suggests an association between lipid transport and an “M2-like” macrophage phenotype

doi: 10.3389/fimmu.2026.1773666

Figure Lengend Snippet: Single cell RNA seq of ccRCC tumors. Data by Bi et al. was visualized and via the Single Cell Portal . Cell types, including immune subpopulations, were annotated as defined by the original authors. Expression of metabolic (ACAA2, SQLE, ACSL3, CD36) and immunologic (CD68, CD163, CD147) genes was examined across these distinct immune compartments. For the characterization of the immune cell populations, please refer to Bi et al. .

Article Snippet: Reagents and antibodies used included FcR Blocking Reagent (Cat# 130-059-901), CD36 PE (Cat# 130-110-877), CD147 APC (Cat# 130-124-295), CD8a PE (Cat# 130-117-201), CD45 PE (Cat# 130-113-118), CD68 PE (Cat# 130-128-345), CD163 PE (Cat# 130-127-908), and pan-Cytokeratin APC (Cat# 130-123-091), all from Miltenyi Biotec.

Techniques: Single Cell, RNA Sequencing, Expressing