Journal: Journal of neuroinflammation

Article Title: Downregulation of CD151 restricts VCAM-1 mediated leukocyte infiltration to reduce neurobiological injuries after experimental stroke.

doi: 10.1186/s12974-021-02171-6

Figure Lengend Snippet: Fig. 7 No direct effect was detected in the activation of neutrophil and monocyte after stimulation by the BMVEC supernatant. a, b In vitro experiment design: the collected BMVEC supernatant was added into the HL-60 cell line or THP-1 culture media and cultured for 72 h. b ROS production in HL-60 was tested. *** vs. LV Vehicle + OGD indicate p < 0.001. c NET formation in HL-60 was detected by ELISA. **** vs. LV Vehicle + OGD indicate p < 0.0001. d ROS production in THP-1 was tested. ** and *** vs. LV Vehicle + OGD indicate p < 0.01 and 0.001, respectively. e sCD14 secretion in THP-1 supernatant was detected by ELISA. * vs. LV Vehicle + OGD indicate p < 0.05. f sCD163 secretion in THP-1 supernatant was detected by ELISA. *** vs. LV Vehicle + OGD indicate p < 0.001

Article Snippet: A human sCD14 ELISA kit and a human sCD163 ELISA kit (Multi Sciences, China) were used to detect THP-1 activation after 72 h of culture with BMVEC supernatant.

Techniques: Activation Assay, In Vitro, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: SUMO modified ETV1 promotes M2-polarized tumor-associated macrophage infiltration and cancer progression by facilitating CCL2 transcription in esophageal squamous cell carcinoma cells

doi: 10.1007/s00262-024-03914-z

Figure Lengend Snippet: CIBERSORT analysis of immune cell infiltration in ESCC samples from TCGA database. A CIBERSORT analysis showed immune cell infiltration and immune cell correlation. B There was macrophage infiltration in ESCC tumor tissues. C TIMER2.0 was used to analyze the correlation between ETV1 and M0 and M2 macrophage infiltration. D Correlation between ETV1 and M2-labeled CD206 or CD163 expression based on the GEPIA database

Article Snippet: The following antibodies were used: human CD68-FITC (65,202, Proteintech, USA); human CD206-PE (65,155, Proteintech); human CD163-PE (65,169, Proteintech); mouse CD206 (12–2061-80, Invitrogen, USA) and mouse CD68 (137,005, Biolegend, USA).

Techniques: Labeling, Expressing

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: SUMO modified ETV1 promotes M2-polarized tumor-associated macrophage infiltration and cancer progression by facilitating CCL2 transcription in esophageal squamous cell carcinoma cells

doi: 10.1007/s00262-024-03914-z

Figure Lengend Snippet: ETV1 expression was significantly elevated in tumor tissues of patients with ESCC. A Results of TCGA and GEPIA databases showed that ETV1 expression was markedly increased in tumor tissues of patients with ESCC. B Real time PCR was used to detect the mRNA expression of ETV1 in fresh frozen ESCC and adjacent samples. C Western blot was employed to determine the protein expression of ETV1 in ESCC and adjacent samples. D The expression of ETV1 in pathological progression of ESCC was tested by immunohistochemical staining. Scale bar, 100 µm. (E–F) The expression of ETV1 and CD206 or CD163 in ESCC tissues was examined by immunofluorescence double staining. Scale bar, 100 µm. The correlation between the fluorescence intensity of ETV1 and the number of CD206 + or CD163 + cells was analyzed. Data are mean ± SD. ****, p < 0.0001

Article Snippet: The following antibodies were used: human CD68-FITC (65,202, Proteintech, USA); human CD206-PE (65,155, Proteintech); human CD163-PE (65,169, Proteintech); mouse CD206 (12–2061-80, Invitrogen, USA) and mouse CD68 (137,005, Biolegend, USA).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Immunohistochemical staining, Staining, Immunofluorescence, Double Staining, Fluorescence

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: SUMO modified ETV1 promotes M2-polarized tumor-associated macrophage infiltration and cancer progression by facilitating CCL2 transcription in esophageal squamous cell carcinoma cells

doi: 10.1007/s00262-024-03914-z

Figure Lengend Snippet: ETV1 facilitates M2 macrophage chemotaxis and polarization. A THP-1-M0 cells were placed in the upper chamber of transwell, and ESCC cells were placed in the lower chamber of transwell. After 24 h of co-culture, the cells migrated to the submembrane surface were detected by crystal violet staining. Scale bar, 100 µm. B , C ESCC cells were placed in the upper chamber of transwell, and THP-1-M0 cells were placed in the lower chamber of transwell. After 24 h of co-culture, the proportions of CD68 + CD206 + and CD68 + CD163 + in THP-1-M0 cells were tested by flow cytometry. Data are mean ± SD. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001

Article Snippet: The following antibodies were used: human CD68-FITC (65,202, Proteintech, USA); human CD206-PE (65,155, Proteintech); human CD163-PE (65,169, Proteintech); mouse CD206 (12–2061-80, Invitrogen, USA) and mouse CD68 (137,005, Biolegend, USA).

Techniques: Chemotaxis Assay, Co-Culture Assay, Staining, Flow Cytometry

Journal: Journal for immunotherapy of cancer

Article Title: EBV promotes TCR-T-cell therapy resistance by inducing CD163+M2 macrophage polarization and MMP9 secretion.

doi: 10.1136/jitc-2023-008375

Figure Lengend Snippet: Figure 1 Increased numbers of exhausted CD8+T cells and CD163+M2 macrophages within EBV-positive tumors. (A) Representative single-stained and merged images of panels (CD8, Granzyme B, DAPI) used for multiplex immunohistochemistry in EBV-positive and EBV-negative tumors. Magnification: ×400 (single-stained images and big merged images), ×200 (small merged images in the bottom right corner). (B–D) Statistical analysis of the density of CD8 (B) or Granzyme B (C) and the ratio of Granzyme B/CD8 (D) in multiplex immunohistochemistry. Mean±SD, n=3, two-tailed t-test. (E) Representative merged images of panels (CD8, Granzyme B, CD163, Pan-CK, DAPI) used in multiplex immunohistochemistry. Magnification: ×400 (small merged images in the top left corner), ×100 (big merged images). (F) Statistical analysis of the density of CD163 in EBV-positive and EBV-negative tumors by multiplex immunohistochemistry. Mean±SD, n=3, two-tailed t-test. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. (G) Correlation analysis between CD163/mm2 and the Granzyme B/CD8 ratio in NPC, GC and LELC. EBV, Epstein-Barr virus; GC, gastric cancer; LELC, lymphoepithelioma-like carcinoma of the lung; NPC, nasopharyngeal cancer.

Article Snippet: After that, 1×106/well CD8+T cells or TCR- T cells were added to the same 24- well plate and cocultured with CD163+M2 macrophages or mononuclear macrophages without polarization for another 3 days with or without 20 μM MMP9 inhibitor (MCE, HY- 135232).

Techniques: Staining, Multiplex Assay, Immunohistochemistry, Two Tailed Test, Virus

Journal: Journal for immunotherapy of cancer

Article Title: EBV promotes TCR-T-cell therapy resistance by inducing CD163+M2 macrophage polarization and MMP9 secretion.

doi: 10.1136/jitc-2023-008375

Figure Lengend Snippet: Figure 3 EBV-infected tumors promote CD163+M2 macrophages polarization. (A) Flow cytometry analysis of the CD163+M2 phenotype in THP-1 or CD14+ monocytes treated with EBV-infected or EBV-uninfected tumor supernatants. Mean±SD, n=4 (HK1), n=3 (AGS), two-tailed t-test. (B) Level of CSF1 in EBV-infected and EBV-uninfected tumor supernatants as measured by ELISA. Mean±SD, n=4, two-tailed t-test. (C) Flow cytometry analysis of the CD163+M2 phenotype of CD14+monocytes treated with 50 ng/mL CSF1, 80 ng/mL IL10, or both together. Mean±SD, n=3, one-way ANOVA. (D) Level of IL10 in EBV- infected and EBV-uninfected tumor supernatants as measured by ELISA. Mean±SD, n=4, two-tailed t-test. (E) Levels of IL10 in supernatants of mononuclear macrophages treated with EBV-infected and EBV-uninfected tumor supernatants for 3 days as measured by ELISA. Mean±SD, n=3, two-tailed t-test. (F) Levels of IL10 in mononuclear macrophage supernatants treated for 3 days with EBV-infected tumor supernatant, EBV-infected tumor supernatant and CSF1R inhibitor, 50 ng/mL CSF1, or no treatment as measured by ELISA. Mean±SD, n=4, two-tailed t-test. (G) Flow cytometry analysis of the CD163+M2 phenotype of CD14+monocytes treated with EBV-infected tumor supernatant (control), EBV-infected tumor supernatant and CSF1R inhibitor, EBV-infected tumor supernatant and anti-IL10R antibody, or EBV-infected tumor supernatant and CSF1R inhibitor and anti- IL10R antibody. Mean±SD, n=3, one-way ANOVA. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. ANOVA, analysis of variance; EBV, Epstein-Barr virus.

Article Snippet: After that, 1×106/well CD8+T cells or TCR- T cells were added to the same 24- well plate and cocultured with CD163+M2 macrophages or mononuclear macrophages without polarization for another 3 days with or without 20 μM MMP9 inhibitor (MCE, HY- 135232).

Techniques: Infection, Flow Cytometry, Two Tailed Test, Enzyme-linked Immunosorbent Assay, Control, Virus

Journal: Journal for immunotherapy of cancer

Article Title: EBV promotes TCR-T-cell therapy resistance by inducing CD163+M2 macrophage polarization and MMP9 secretion.

doi: 10.1136/jitc-2023-008375

Figure Lengend Snippet: Figure 4 CD163+M2 macrophages polarized by EBV suppresses T-cell function. (A) Volcano plot of differentially expressed genes between mononuclear macrophages induced with EBV-uninfected tumor supernatant and those induced with EBV- infected tumor supernatant for 3 days. (B) GO pathway enrichment analysis of differentially expressed genes between mononuclear macrophages treated with EBV-infected and EBV-uninfected tumor supernatant. (C, D) GSEA of differentially expressed genes between mononuclear macrophages treated with EBV-infected and EBV-uninfected tumor supernatant. (E) Flow cytometry analysis of T-cell functional molecules (Granzyme B, IFN γ, TNF α, Perforin) after 3 days of coculture with mononuclear macrophages polarized with EBV-uninfected or EBV-infected tumor supernatants. Mean±SD, n=3, two-tailed t-test. (F) Flow cytometry analysis of T-cell functional molecules after 3 days of coculture with mononuclear macrophages untreated (control) or polarized with 50 ng/mL CSF1+80 ng/mL IL10 (M2). (G) Statistical analysis of T-cell functional molecules in (F). Mean±SD, n=3, two-tailed t-test. (H) Flow cytometry analysis of T-cell functional molecules after 3 days of coculture with mononuclear macrophages induced by EBV-infected tumor culture medium (control), EBV-infected tumor culture medium and CSF1R inhibitor (CSF1Rin), EBV-infected tumor culture medium and anti-IL10R antibody (anti-IL10) or EBV-infected tumor culture medium and CSF1R inhibitor and anti-IL10R antibody (CSF1Rin+anti-IL10). (I) Statistical analysis of T-cell functional molecules in (H). Mean±SD, n=3, one-way ANOVA. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. ANOVA, analysis of variance; EBV, Epstein-Barr virus; GO, Gene Ontology; GSEA, gene set enrichment analysis.

Article Snippet: After that, 1×106/well CD8+T cells or TCR- T cells were added to the same 24- well plate and cocultured with CD163+M2 macrophages or mononuclear macrophages without polarization for another 3 days with or without 20 μM MMP9 inhibitor (MCE, HY- 135232).

Techniques: Cell Function Assay, Infection, Flow Cytometry, Functional Assay, Two Tailed Test, Control, Virus

Journal: Journal for immunotherapy of cancer

Article Title: EBV promotes TCR-T-cell therapy resistance by inducing CD163+M2 macrophage polarization and MMP9 secretion.

doi: 10.1136/jitc-2023-008375

Figure Lengend Snippet: Figure 5 EBV-polarized CD163+M2 macrophages inhibit the cytotoxic capacity of TCR-T cells against tumors. (A) Flow cytometry analysis of functional molecules in TCR-T cells cocultured with CSF1+IL10-polarized M2 (M2) or untreated mononuclear macrophages (control) for 3 days. Mean±SD, n=4, two-tailed t-test. (B) Left: Images of C666-1-A11-LMP2A cells killed by TCR-T cells after coculture with untreated mononuclear macrophages (control treated TCR-T) or CSF1+IL10-polarized M2 macrophages (M2 treated TCR-T) for 3 days and then isolated via MACS at 0 hour and 16 hours. Magnification: ×100. Right: Statistical analysis of LDH released from C666-1-A11-LMP2A cells after 16 hours of killing by TCR-T cells. Mean±SD, n=3, two-tailed t-test. (C) Apoptosis rate analysis of C666-1-A11-LMP2A cells killed by TCR-T cells with different treatment for 16 hours by flow cytometry. Mean±SD, n=6, two-tailed t-test. (D) Experimental scheme diagram for the subcutaneous xenograft tumor model in NCG mice. (E) Tumor volume (left) and tumor weight (right) at day 26 after implantation. Mean±SD, n=7 (M2, M2+TCR T), n=8 (TCR-T, control), two-tailed t-test. (F) Left: Serial sections of mouse xenografts were stained with HE and antibodies targeting CD163, CD3, and Granzyme B. Magnification: ×200. Right: Statistical analysis of the number of CD163+cells, CD3+cells, and Granzyme B. Mean±SD, n=3, two-tailed t-test or Mann-Whitney U test. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. EBV, Epstein-Barr virus; LDH, lactate dehydrogenase; HE, hematoxylin-eosin.

Article Snippet: After that, 1×106/well CD8+T cells or TCR- T cells were added to the same 24- well plate and cocultured with CD163+M2 macrophages or mononuclear macrophages without polarization for another 3 days with or without 20 μM MMP9 inhibitor (MCE, HY- 135232).

Techniques: Flow Cytometry, Functional Assay, Control, Two Tailed Test, Isolation, Staining, MANN-WHITNEY, Virus

Journal: Journal for immunotherapy of cancer

Article Title: EBV promotes TCR-T-cell therapy resistance by inducing CD163+M2 macrophage polarization and MMP9 secretion.

doi: 10.1136/jitc-2023-008375

Figure Lengend Snippet: Figure 6 MMP9 secreted by EBV-induced CD163+M2 macrophages suppresses TCR-T-cell function in vitro. (A) Levels of MMP9 in the supernatants of mononuclear macrophages treated with different conditions (left) as well as in EBV-infected or EBV-uninfected tumor supernatants (right) measured by ELISA. Mean±SD, n=4, two-tailed t-test. (B) Left: Representative images of EBV-positive or EBV-negative nasopharyngeal and gastric cancer tissues stained with MMP9 antibody. Magnification: ×200. Right: IHC scores of MMP9. Mean±SD, n=5 (GC and NPC+), n=4 (NPC−), two-tailed t-test. (C) Flow cytometry analysis of functional molecules in TCR-T cells after 3 days of coculture with M2 macrophages induced by CSF1+IL10, M2 macrophages and MMP9 inhibitors, untreated monocytes (control), or untreated monocytes and MMP9 inhibitors (without C666-1-A11- LMP2A). Mean±SD, n=4, one-way ANOVA. (D) Images of C666-1-A11-LMP2A cells killed by TCR-T cells at 0 hour and 16 hours which were cocultured with M2 macrophages, M2 macrophages+MMP9 inhibitor, untreated monocytes or untreated monocytes+MMP9 inhibitor for 3 days and then isolated via MACS. Magnification: ×100. (E) Statistical analysis of LDH released from C666-1-A11-LMP2A cells after 16 hours of killing by TCR-T cells with different treatment. Mean±SD, n=5, two-tailed t-test. (F) Apoptosis analysis of C666-1-A11-LMP2A cells killed by TCR-T cells for 16 hours which were cocultured with M2 macrophages, M2 macrophages+MMP9 inhibitor, untreated monocytes or untreated monocytes+MMP9 inhibitor for 3 days and then isolated via MACS Mean±SD, n=4, two-tailed t-test. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. ANOVA, analysis of variance; EBV, Epstein-Barr virus; IHC, immunohistochemistry.

Article Snippet: After that, 1×106/well CD8+T cells or TCR- T cells were added to the same 24- well plate and cocultured with CD163+M2 macrophages or mononuclear macrophages without polarization for another 3 days with or without 20 μM MMP9 inhibitor (MCE, HY- 135232).

Techniques: Cell Function Assay, In Vitro, Infection, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Staining, Flow Cytometry, Functional Assay, Control, Isolation, Virus, Immunohistochemistry

Journal: Clinical and Experimental Pharmacology & Physiology

Article Title: Mechanisms of IL‐17A Neutralisation in Alleviating Renal Fibrosis and Inflammation in Spontaneously Hypertensive Rats

doi: 10.1111/1440-1681.70116

Figure Lengend Snippet: Effects of IL‐17A neutralisation on macrophage polarisation in SHR renal tissues. (A) Representative IHC staining of M1 macrophage markers (iNOS and CD86) with quantitative analysis of their positive areas. (B) Representative IHC staining of M2 macrophage markers (Arg‐1 and CD163) with quantitative analysis of their positive areas. (C) Representative immunoblots and relative expression levels of iNOS, CD86, Arg‐1, and CD163 proteins. (D) Proportion of CD86 + and CD163 + cells among CD68 + macrophages. (E) mRNA expression levels of iNOS, CD86, Arg‐1, and CD163. Data are presented as mean ± SD ( n = 6).

Article Snippet: Sections were then incubated overnight at 4°C with primary antibodies against: E‐cadherin (Boster, China), Collagen III (Boster, China), inducible nitric oxide synthase (iNOS) (Boster, China), CD86 (Boster, China), arginase‐1 (Arg‐1) (Boster, China), CD163 (Boster, China), α‐smooth muscle actin (α‐SMA) (Cell Signalling Technology, USA).

Techniques: Immunohistochemistry, Western Blot, Expressing

Journal: Experimental and Therapeutic Medicine

Article Title: Role of SARS‑CoV‑2 nucleocapsid protein in affecting immune cells and insights on its molecular mechanisms

doi: 10.3892/etm.2023.12203

Figure Lengend Snippet: Quantification of TNF-α, IL-10, IL-6 and CD163. The concentrations of (A) TNF-α, (B) IL-10, (C) IL-6 and (D) CD163 were determined in the supernatant of cultured Raw264.7 cells treated with the severe acute respiratory syndrome coronavirus 2 N protein. Data are presented as mean ± SD (n=3). * P<0.05 and ** P<0.01 vs. blank control.

Article Snippet: PageRuler TM Prestained protein ladder (cat. no. 26616) was purchased from Thermo Fisher Scientific, Inc. Trypsin-EDTA (0.25%) used to digest Raw264.7 cells for cell passage was purchased from Gibco; Thermo Fisher Scientific Inc. TNF-α Elisa kit (Cat. no. EK0527), IL-6 Elisa Kit (Cat. no. EK0411), IL-10 Elisa Kit (Cat. no. EK0417) and CD163 ELISA kit (Cat. no. EK1146) were purchased from Boster Biological Technology co., Ltd.).

Techniques: Cell Culture, Control

Journal: Open Forum Infectious Diseases

Article Title: High-Density Lipoprotein-Mediated Cholesterol Efflux Capacity Is Improved by Treatment With Antiretroviral Therapy in Acute Human Immunodeficiency Virus Infection

doi: 10.1093/ofid/ofu108

Figure Lengend Snippet: Baseline Characteristics by Randomization Group a

Article Snippet: Plasma soluble CD163 (sCD163), monocyte chemoattractant protein-1 (MCP-1), interleukin (IL)-6, and sCD14 levels were quantified by enzyme-linked immunosorbent assay (ELISA) (Trillium Diagnostics [sCD163] and R&D Systems [high-sensitivity IL-6, MCP-1, and sCD14]).

Techniques: Activation Assay, Translocation Assay

Journal: Open Forum Infectious Diseases

Article Title: High-Density Lipoprotein-Mediated Cholesterol Efflux Capacity Is Improved by Treatment With Antiretroviral Therapy in Acute Human Immunodeficiency Virus Infection

doi: 10.1093/ofid/ofu108

Figure Lengend Snippet: Changes From Baseline to 12 Weeks in Both Randomization Groups (Untreated vs Antiretroviral Treatment Group) a

Article Snippet: Plasma soluble CD163 (sCD163), monocyte chemoattractant protein-1 (MCP-1), interleukin (IL)-6, and sCD14 levels were quantified by enzyme-linked immunosorbent assay (ELISA) (Trillium Diagnostics [sCD163] and R&D Systems [high-sensitivity IL-6, MCP-1, and sCD14]).

Techniques: Activation Assay, Translocation Assay

Journal: Open Forum Infectious Diseases

Article Title: High-Density Lipoprotein-Mediated Cholesterol Efflux Capacity Is Improved by Treatment With Antiretroviral Therapy in Acute Human Immunodeficiency Virus Infection

doi: 10.1093/ofid/ofu108

Figure Lengend Snippet: Correlation Between Change in CV Risk Factors, HIV Disease-related Parameters, and Markers of Monocyte/Macrophage Activation and Inflammation to the Change in Abca1 +/+ Efflux Capacity Among All Subjects (ART and Non-ART Treated)

Article Snippet: Plasma soluble CD163 (sCD163), monocyte chemoattractant protein-1 (MCP-1), interleukin (IL)-6, and sCD14 levels were quantified by enzyme-linked immunosorbent assay (ELISA) (Trillium Diagnostics [sCD163] and R&D Systems [high-sensitivity IL-6, MCP-1, and sCD14]).

Techniques: Activation Assay, Translocation Assay, Marker

Journal: Open Forum Infectious Diseases

Article Title: High-Density Lipoprotein-Mediated Cholesterol Efflux Capacity Is Improved by Treatment With Antiretroviral Therapy in Acute Human Immunodeficiency Virus Infection

doi: 10.1093/ofid/ofu108

Figure Lengend Snippet: Multivariate Model to Assess Relationships to Change in Abca1 +/+ Cholesterol Efflux (%)

Article Snippet: Plasma soluble CD163 (sCD163), monocyte chemoattractant protein-1 (MCP-1), interleukin (IL)-6, and sCD14 levels were quantified by enzyme-linked immunosorbent assay (ELISA) (Trillium Diagnostics [sCD163] and R&D Systems [high-sensitivity IL-6, MCP-1, and sCD14]).

Techniques: