Journal: Journal of Leukocyte Biology

Article Title: The chemotaxis of M1 and M2 macrophages is regulated by different chemokines

doi: 10.1189/jlb.1a0314-170r

Figure Lengend Snippet: Figure 1. Schematic diagram of screening of chemokines in M1/M2 macrophage chemotaxis. Forty-one chemokines, ligands of all 20 chemokine receptors, were subjected to chemotaxis assay by use of M1 and M2 macrophages derived from human CD14+ monocytes from six donors. To confirm the screening results, 13 hits were subjected to full dose-response chemotaxis assay by use of M1 and M2 macrophages derived from human CD14+ monocytes.

Article Snippet: Human monocytes were purified from normal PBMCs by magnetic cell sorting by use of CD14 microbeads (Miltenyi Biotec, Bergisch Gladbach, Forty-one chemokines, ligands of all 20 chemokine receptors, were subjected to chemotaxis assay by use of M1 and M2 macrophages derived from human CD14+ monocytes from six donors.

Techniques: Chemotaxis Assay, Derivative Assay

Journal: Journal of Leukocyte Biology

Article Title: The chemotaxis of M1 and M2 macrophages is regulated by different chemokines

doi: 10.1189/jlb.1a0314-170r

Figure Lengend Snippet: Figure 2. Dose-dependent effect of chemokines on human M1 and M2 macrophage chemotaxis. M1 and M2 macrophages derived from CD14+

Article Snippet: Human monocytes were purified from normal PBMCs by magnetic cell sorting by use of CD14 microbeads (Miltenyi Biotec, Bergisch Gladbach, Forty-one chemokines, ligands of all 20 chemokine receptors, were subjected to chemotaxis assay by use of M1 and M2 macrophages derived from human CD14+ monocytes from six donors.

Techniques: Chemotaxis Assay, Derivative Assay

Journal: Journal of Leukocyte Biology

Article Title: The chemotaxis of M1 and M2 macrophages is regulated by different chemokines

doi: 10.1189/jlb.1a0314-170r

Figure Lengend Snippet: Figure 3. Expression of chemokine receptors in human M1 and M2 macrophages. (A) Relative mRNA levels of chemokine receptors in M1 and M2 macrophages derived from human CD14+ monocytes were assessed by use of qRT-PCR. Results were normalized with the level of GAPDH. Data are mean 6 SEM from 8 independent experiments. (B) Representative Western blots showing the expression of CCR7, CCR3, and CXCR4 in the whole cell lysate of M1 and M2 macrophages derived from CD14+ monocytes. b-Actin was used as loading control. (C) Quantification of the expression of CCR7, CCR3, and CXCR4 in the whole cell lysates of M1 and M2 macrophages. Data were mean 6 SEM from 3 independent experiments. *P , 0.05, **P , 0.01, ***P , 0.001 compared with their expression levels in M1 macrophages. (D) Representative Western blot showing predominant expression of CCR7 in the membrane fraction (M) of M1 and the cytosolic fraction (C) of M2 macrophages derived from CD14+ monocytes. Na, K-ATPase was used as loading control for membrane fraction, and a-tubulin was used as loading control for cytoplasmic fraction.

Article Snippet: Human monocytes were purified from normal PBMCs by magnetic cell sorting by use of CD14 microbeads (Miltenyi Biotec, Bergisch Gladbach, Forty-one chemokines, ligands of all 20 chemokine receptors, were subjected to chemotaxis assay by use of M1 and M2 macrophages derived from human CD14+ monocytes from six donors.

Techniques: Expressing, Derivative Assay, Quantitative RT-PCR, Western Blot, Control, Membrane

Journal: Journal of Leukocyte Biology

Article Title: The chemotaxis of M1 and M2 macrophages is regulated by different chemokines

doi: 10.1189/jlb.1a0314-170r

Figure Lengend Snippet: Figure 5. CCL19- or CCL21-induced M1 macro- phage chemotaxis was blocked by a neutralizing antibody against CCR7. M1 or M2 macrophages derived from human CD14+ monocytes were subjected to chemotaxis analysis in the presence of CCL19 or CCL21 (10 nM) and either a CCR7 neutralizing antibody (30 mg/ml) or an isotype (IgG2a) control. Data were mean chemotactic index 6 SEM from 5 independent experiments. *P , 0.05, **P , 0.01 compared with control (vehicle treatment).

Article Snippet: Human monocytes were purified from normal PBMCs by magnetic cell sorting by use of CD14 microbeads (Miltenyi Biotec, Bergisch Gladbach, Forty-one chemokines, ligands of all 20 chemokine receptors, were subjected to chemotaxis assay by use of M1 and M2 macrophages derived from human CD14+ monocytes from six donors.

Techniques: Chemotaxis Assay, Derivative Assay, Control

Journal: Journal of Leukocyte Biology

Article Title: The chemotaxis of M1 and M2 macrophages is regulated by different chemokines

doi: 10.1189/jlb.1a0314-170r

Figure Lengend Snippet: Figure 7. CCL19- or CCL21-induced chemotaxis of M1 macrophages was blocked by PI3K inhibitor rather than MEK1 inhibitor. M1 macrophages derived from human CD14+ monocytes (n = 5) were pretreated with Wortmannin (3 mM) or PD98059 (30 mM) before being subjected to chemotaxis analysis in the presence of 10 nM CCL19 (A) or 10 nM CCL21 (B). Data were mean chemotactic index 6 SEM. *P , 0.05, ***P , 0.001 compared with control (vehicle treatment).

Article Snippet: Human monocytes were purified from normal PBMCs by magnetic cell sorting by use of CD14 microbeads (Miltenyi Biotec, Bergisch Gladbach, Forty-one chemokines, ligands of all 20 chemokine receptors, were subjected to chemotaxis assay by use of M1 and M2 macrophages derived from human CD14+ monocytes from six donors.

Techniques: Chemotaxis Assay, Derivative Assay, Control

Journal: Bioactive Materials

Article Title: Recruited CD68 + CD206 + macrophages orchestrate graft immune tolerance to prompt xenogeneic-dentin matrix-based tooth root regeneration

doi: 10.1016/j.bioactmat.2020.09.029

Figure Lengend Snippet: M2γ macrophages suppressed the Th1-type CTL response triggered by xeno-complex in vitro . (A) The hypothesis that polarized M2 macrophages perform anti-inflammatory effects on cellular response triggered by xeno-complex. (B) Incubation and identification of PPARγ-primed macrophages (M2γ). Human peripheral monocytes were primed towards M2γ macrophages via the addition of rosiglitazone (RSG) in the presence of IL-4. The M2γ macrophages were obtained after incubation for 7 days. The phenotype marker CD206 was identified by Flow cytometry analysis (FACS). Data are means ± SEM (n ≥ 3). * p < 0.05. (C) Identification of M2γ conditional medium (M2γ medium). The anti-inflammatory and pro-inflammatory cytokines were analyzed using Immunology Multiplex MAP. Data are means ± SEM (n ≥ 3). * p < 0.05, *** p < 0.001. (D) M2γ medium or RSG inhibited proliferated CD3 + CD8 + cells triggered by xeno-complex. The cellular response to xeno-complex was quantified by FCS using CFSE-based proliferation assay. The proliferated CD3 + , CD3 + CD4 + , and CD3 + CD8 + lymphocytes were graphed as an overlay (black line) in each FACS histogram covering hPBMCs (gray shaded histogram). hPBMCs without stimulus was the negative group, and PHA stimulation was the positive group. The % value on the left defined the level of proliferated cells within 5 days. Data are means ± SEM (n ≥ 3). * p < 0.05, ** p < 0.01. (E) hPBMCs in the six coculture systems were measured for mRNA expression of pro-inflammatory TNF-α, INF-γ. The data were normalized to those of GAPDH mRNA and are presented relative to those of hPBMCs without any stimulus, set as 1. Data are means ± SEM (n ≥ 3). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; NS, no significance. (F) The adherent monocytes/macrophages differentiation in the six coculture systems were co-immunostained with antibodies against CD68/CCR7 or CD68/CD206, DAPI was used for nuclear staining. Scale bars, 50 μm.

Article Snippet: The following primary antibodies were used: the pan-macrophage marker, mouse anti-rat CD68 (Abcam, Cambridge, MA, UK), the M1 macrophage marker, rabbit anti-rat CCR7 (Abcam, Cambridge, MA, UK), and the M2 macrophage marker, goat anti-rat CD206 (Santa Cruz).

Techniques: In Vitro, Incubation, Marker, Flow Cytometry, Multiplex Assay, Proliferation Assay, Expressing, Staining

Journal: Bioactive Materials

Article Title: Recruited CD68 + CD206 + macrophages orchestrate graft immune tolerance to prompt xenogeneic-dentin matrix-based tooth root regeneration

doi: 10.1016/j.bioactmat.2020.09.029

Figure Lengend Snippet: PPARγ activation locally modulated M1-to-M2 tissue macrophage polarization. (A) The schematic diagram illustrated the correlation between M1/M2 macrophage phenotypes and the fate of implanted ECM. Macrophage polarization in response to xeno-complex were sequentially observation at postoperative 3 days, 1 week, 3 weeks, 6 weeks, and 12 weeks. Xeno-complex with RSG treatment served as experimental groups, while with no treatment (Control) or with vehicle (Vehicle control) served as controls. Macrophage phenotypes (B) at periphery sites and (C) in the intracavity sites of xeno-complex were shown by representative confocal immunofluorescence images. Co-immunostaining of pan-macrophage cell surface marker CD68 (red), the M1 marker CCR7 (orange), and the M2 marker CD206 (green) were merged with nuclear staining DAPI (blue). Scale bars, 50 μm. (D) A ratio was calculated as to the percentage of M2 cells to M1 cells (M2/M1 radio). The gray dotted line represented M2/M1 radio as 1.0, with over 1.0 indicating predominant M2 vs. below 1.0 representing predominant M1 cells. Data are mean ± SEM (n ≥ 3), * p < 0.05, ** p < 0.01, **** p < 0.0001. NS, no significance. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The following primary antibodies were used: the pan-macrophage marker, mouse anti-rat CD68 (Abcam, Cambridge, MA, UK), the M1 macrophage marker, rabbit anti-rat CCR7 (Abcam, Cambridge, MA, UK), and the M2 macrophage marker, goat anti-rat CD206 (Santa Cruz).

Techniques: Activation Assay, Control, Immunofluorescence, Immunostaining, Marker, Staining

Journal: Life sciences

Article Title: The impact of ExHp-CD (outer membrane vesicles) released from Helicobacter pylori SS1 on macrophage RAW 264.7 cells and their immunogenic potential.

doi: 10.1016/j.lfs.2021.119644

Figure Lengend Snippet: Fig. 8. Immunofluorescence staining of macrophage RAW 264.7 cells treated with 5 and 10 μg/mL of ExHp-CD. M2 marker CD206 (A) and M1 marker (B) are shown in green and blue for cell nuclei (DAPI). Compared to the control, the groups stimulated with 5 and 10 μg/mL of ExHp-CD showed an increased ratio of CD206 and a decreased ratio of CD86. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The primary antibodies against the M1 marker CD86 (1:100, CD86 polyclonal antibody, Proteintech, Rosemont, IL 60018, USA) and the M2 marker CD206 (1:100, CD206 polyclonal antibody, Proteintech, Rosemont, IL 60018, USA) were used.

Techniques: Immunofluorescence, Staining, Marker, Control

Journal: Breast Cancer Research : BCR

Article Title: AXL promotes inflammatory breast cancer progression by regulating immunosuppressive macrophage polarization

doi: 10.1186/s13058-025-02015-8

Figure Lengend Snippet: AXL inhibition suppresses the polarization of immunosuppressive M2 macrophages. A AXL was knocked out in human THP-1 monocytes using the CRISPR/Cas9 system. These cells and control cells were subsequently induced into M2 macrophages, and AXL depletion was confirmed in AXL-KO M2 macrophages using Western blotting. M2 macrophages polarized from AXL-KO THP-1 cells had lower expression of AXL than M2 macrophages polarized from control cells. B The expression of M2 macrophage markers and secreted cytokines was measured by qRT-PCR. AXL KO reduced the expression of CD163 , CD206 , CCL17 , and CCL18 in M2 macrophages. C Flow cytometric analysis was conducted to compare the CD206 + macrophage population polarized from AXL-KO versus control THP-1 cells. AXL KO reduced the population of CD206 + macrophages polarized from THP-1 cells. D Treatment with TP-0903 inhibited the expression of AXL mRNA (left panel) and protein (right panel) in THP-1–derived M2 macrophages as determined using qRT-PCR and Western blotting, respectively. E TP-0903–treated M2 macrophages showed reduced mRNA level of CD163 , CD206 , CCL17 , and CCL18 compared to vehicle-treated M2 macrophages. F Flow cytometry and Western blotting showed that TP-0903 reduced the population of CD206 + cells (left panel) and CD206 protein expression (right panel) in THP-1–derived M2 macrophages. All experiments were repeated at least three times. Data were summarized as means ± SD. A 2-tailed Student t test ( B and 1-way analysis of variance followed by Dunnett’s multiple comparison test ( D – F ) were used to calculate P values. * P < 0.01

Article Snippet: Protein levels of human CCL20, CCL26, EREG, CXCL9, and CXCL10 in lysates or CM from AXL-KO or control M2 macrophages were quantified using a DuoSet ELISA kit, following the manufacturer’s instructions (R&D Systems).

Techniques: Inhibition, CRISPR, Control, Western Blot, Expressing, Quantitative RT-PCR, Derivative Assay, Flow Cytometry, Comparison

Journal: Breast Cancer Research : BCR

Article Title: AXL promotes inflammatory breast cancer progression by regulating immunosuppressive macrophage polarization

doi: 10.1186/s13058-025-02015-8

Figure Lengend Snippet: AXL depletion reduces the impact of M2 macrophages on IBC cell growth and migration. A SUM149 and BCX010 cells were co-cultured with 100% CM collected after culturing vehicle- or TP-0903–treated M2 macrophages for 48 h, and cell numbers after 3 days were measured by CTB assay. CM from TP-0903–treated M2 macrophages reduced the growth of SUM149 and BCX010 IBC cells. B The migration of human IBC cells induced by 100% CM from TP-0903– or vehicle-treated M2 macrophages after 48 h of culture was examined using transwell migration assay. CM from TP-0903–treated M2 macrophages inhibited the migration of SUM149 and BCX010 cells ( C ). D 100% CM from AXL-KO M2 macrophages after 48 h of culture reduced the growth C and migration D of SUM149 and BCX010 cells. All experiments were repeated at least three times. Data were summarized as means ± SD. One-way analysis of variance followed by Dunnett’s multiple comparison test ( A and B ) and 2-tailed Student t test ( C and D ) were used to calculate P values. * P < 0.05; ** P < 0.01

Article Snippet: Protein levels of human CCL20, CCL26, EREG, CXCL9, and CXCL10 in lysates or CM from AXL-KO or control M2 macrophages were quantified using a DuoSet ELISA kit, following the manufacturer’s instructions (R&D Systems).

Techniques: Migration, Cell Culture, CtB Assay, Transwell Migration Assay, Comparison

Journal: Breast Cancer Research : BCR

Article Title: AXL promotes inflammatory breast cancer progression by regulating immunosuppressive macrophage polarization

doi: 10.1186/s13058-025-02015-8

Figure Lengend Snippet: AXL suppression inhibits the polarization of immunosuppressive M2 macrophages via STAT6. A Treatment with TP-0903 reduced AXL, phospho-STAT6, and STAT6 protein expression as determined by Western blotting. B M2 macrophages polarized from AXL-KO THP-1 cells had lower phospho-AXL, AXL, phospho-STAT6, and STAT6 protein expression than those polarized from control THP-1 cells, as determined using Western blotting. C–E STAT6 was knocked down in THP-1 cells using siRNAs, and then THP-1 cells were induced to M2 macrophages. Knockdown of STAT6 in THP-1–polarized M2 macrophages C decreased the CD163 + CD206 + macrophage population as determined by flow cytometry D and decreased the expression of the CD163 and CD206 genes as determined using qRT-PCR E . F STAT6 was overexpressed in M2 macrophages polarized from AXL-KO THP-1 cells as tested using qRT-PCR. G STAT6 overexpression mitigated the inhibitory effect of AXL KO on the expression of M2 macrophage markers and cytokines, including CD163 , CD206 , CCL17 , and CCL18 . H The CM from control, AXL-KO, and AXL-KO + STAT6–overexpressing M2 macrophages and fresh media were used as attractants plated in the bottom chamber of transwells to test the migration of SUM149 cells. Migration of SUM149 cells was greater with CM from AXL-KO + STAT6–overexpressing M2 macrophages than with CM from AXL-KO M2 macrophages. All experiments were repeated at least three times. Data were summarized as means ± SD. One-way analysis of variance followed by Dunnett’s multiple comparison test was used to calculate P values. * P < 0.05; ** P < 0.01

Article Snippet: Protein levels of human CCL20, CCL26, EREG, CXCL9, and CXCL10 in lysates or CM from AXL-KO or control M2 macrophages were quantified using a DuoSet ELISA kit, following the manufacturer’s instructions (R&D Systems).

Techniques: Expressing, Western Blot, Control, Knockdown, Flow Cytometry, Quantitative RT-PCR, Over Expression, Migration, Comparison

Journal: Breast Cancer Research : BCR

Article Title: AXL promotes inflammatory breast cancer progression by regulating immunosuppressive macrophage polarization

doi: 10.1186/s13058-025-02015-8

Figure Lengend Snippet: AXL regulates the expression of cytokines via STAT6 in M2 macrophages. A – C mRNA was collected from M2 macrophages polarized from AXL-KO THP-1 and control cells, and the expression of M2 macrophage markers or cytokines/chemokines was examined using qRT-PCR. AXL KO in M2 macrophages derived from THP-1 cells reduced the mRNA expression of CD209 , IL13RA , and IL2RG A . AXL KO in M2 macrophages derived from THP-1 cells reduced the expression of the immunosuppressive cytokines/chemokines CCL20 , CCL26 , EREG , and IL1B B . AXL KO in M2 macrophages derived from THP-1 cells increased the expression of cytokines/chemokines involved in the interferon γ–mediated signaling pathway, such as IFNG , CXCL10 , and GBP2 , at the gene level C . D and E CM from and lysates of M2 macrophages polarized from AXL-KO THP-1 cells had decreased expression of CCL20, CCL26, and EREG protein D but increased expression of CXCL9 and CXCL10 protein E as determined using ELISA. F The migration of human IBC cells was assessed using a transwell migration assay, with CM collected from control and AXL-KO M2 macrophages with or without the addition of recombinant CCL20, CCL26, and EREG protein, serving as attractants. CCL20, CCL26, and EREG mitigated the inhibitory effect of AXL-KO M2 macrophages on SUM149 and BCX010 cell migration. G qRT-PCR was conducted to measure the mRNA expression level of CCL20 , CCL26 , and EREG in control, AXL-KO, and AXL-KO + STAT6-overexpressing M2 macrophages. Overexpression of STAT6 mitigated the suppressive effect of AXL KO in M2 macrophages on the expression of CCL20 , CCL26 , and EREG genes. All experiments were repeated at least three times. Data were summarized as means ± SD. Two-tailed Student t test ( A – E ) and 1-way analysis of variance followed by Dunnett’s multiple comparison test ( F and G ) were used to calculate P values. * P < 0.05; ** P < 0.01; *** P < 0.001

Article Snippet: Protein levels of human CCL20, CCL26, EREG, CXCL9, and CXCL10 in lysates or CM from AXL-KO or control M2 macrophages were quantified using a DuoSet ELISA kit, following the manufacturer’s instructions (R&D Systems).

Techniques: Expressing, Control, Quantitative RT-PCR, Derivative Assay, Enzyme-linked Immunosorbent Assay, Migration, Transwell Migration Assay, Recombinant, Over Expression, Two Tailed Test, Comparison

Journal: Breast Cancer Research : BCR

Article Title: AXL promotes inflammatory breast cancer progression by regulating immunosuppressive macrophage polarization

doi: 10.1186/s13058-025-02015-8

Figure Lengend Snippet: AXL expression correlates with an immunosuppressive TME of IBC. A – D Violin plots showed the absolute percentages of different immune cell subsets as defined by CIBERSORT according to high versus low AXL expression in IBC samples from the Inflammatory Breast Cancer International Consortium: M2 macrophages A , resting memory CD4 + T cells B , activated myeloid dendritic cells C , and follicular helper T cells D . A 2-tailed Student t test was used to calculate P values. E Proposed mechanistic model of AXL in the IBC TME. AXL regulated M2 macrophage polarization and the expression of immunosuppressive molecules and STAT6-modulated cytokines, which induced IBC’s aggressiveness

Article Snippet: Protein levels of human CCL20, CCL26, EREG, CXCL9, and CXCL10 in lysates or CM from AXL-KO or control M2 macrophages were quantified using a DuoSet ELISA kit, following the manufacturer’s instructions (R&D Systems).

Techniques: Expressing

Journal: Arthritis & Rheumatology (Hoboken, N.j.)

Article Title: Activated M2 Macrophages Contribute to the Pathogenesis of IgG4‐Related Disease via Toll‐like Receptor 7/Interleukin‐33 Signaling

doi: 10.1002/art.41052

Figure Lengend Snippet: Identification of TLR ‐7–expressing cells in SMG s from patients with IgG4‐ RD and in normal secondary lymphoid organs from patients with oral squamous cell carcinoma. A , Staining of serial sections of normal tonsil, normal lymph node, and IgG4‐ RD SMG s with H&E, and for TLR ‐7, CD 68 as a marker of both M1 and M2 macrophages, CD 163 as a marker of M2 macrophages, CD 11c as a marker of myeloid dendritic cells, and CD 123 as a marker of plasmacytoid dendritic cells. Outlined areas indicate GC s. Mayer's hematoxylin (blue) counterstained; bars = 100 μm. B , Double immunostaining for TLR ‐7 (red), CD 123 (green), and CD 163 (green) in normal tonsil and SMG s from representative patients with IgG4‐ RD . DAPI (blue) counterstained; bars = 50 μm. See Figure for definitions.

Article Snippet: Cells (5 × 10 6 cells/ml) were cultured using a CellXVivo Human M2 Macrophage Differentiation Kit (R&D Systems) for 6 days in a humidified chamber with 5% CO 2 at 37°C.

Techniques: Expressing, Staining, Marker, Double Immunostaining

Journal: Arthritis & Rheumatology (Hoboken, N.j.)

Article Title: Activated M2 Macrophages Contribute to the Pathogenesis of IgG4‐Related Disease via Toll‐like Receptor 7/Interleukin‐33 Signaling

doi: 10.1002/art.41052

Figure Lengend Snippet: Expression of interleukin‐33 ( IL ‐33) and candidate TLR s in SG s from patients with IgG4‐ RD . A , Expression levels of mRNA for IL ‐33 in SG s from healthy controls (n = 10), patients with chronic sialadenitis (n = 10), patients with SS (n = 15), and patients with IgG4‐ RD (n = 15). B , Distribution of IL ‐33 in SG s from a representative healthy control, patient with chronic sialadenitis, patient with SS , and patient with IgG4‐ RD . Mayer's hematoxylin (blue) counterstained; bars = 100 μm. C , Correlation between expression levels of IL ‐33 mRNA and candidate TLR s in SG s from patients with IgG4‐ RD (n = 15), as determined by Spearman's rank correlation test. D , Schematic illustration of the extraction of CD 163+ M2 macrophages stimulated with TLR ‐7 agonist R848. Cells were cultivated as described in Patients and Methods. PBMC = peripheral blood mononuclear cell. E , Production of IL ‐33 by CD 163+ M2 macrophages stimulated with R848, as determined by enzyme‐linked immunosorbent assay. IL ‐33 levels increased in a concentration‐dependent manner. In A and E , bars show the mean ± SD . * = P < 0.05; ** = P < 0.01 by Kruskal‐Wallis test. See Figure for other definitions.

Article Snippet: Cells (5 × 10 6 cells/ml) were cultured using a CellXVivo Human M2 Macrophage Differentiation Kit (R&D Systems) for 6 days in a humidified chamber with 5% CO 2 at 37°C.

Techniques: Expressing, Control, Extraction, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Arthritis & Rheumatology (Hoboken, N.j.)

Article Title: Activated M2 Macrophages Contribute to the Pathogenesis of IgG4‐Related Disease via Toll‐like Receptor 7/Interleukin‐33 Signaling

doi: 10.1002/art.41052

Figure Lengend Snippet: Schematic model of the role of Toll‐like receptor 7 ( TLR ‐7)–positive M2 macrophages in the initiation of Ig4‐related disease. TLR ‐7 expressed on M2 macrophages recognize some RNA viruses or self RNA from apoptotic cells. Activated M2 macrophages secrete interleukin‐33 ( IL ‐33) and promote production of Th2 cytokines that lead to IgG4 class‐switching and fibrosis. TGF β = transforming growth factor β.

Article Snippet: Cells (5 × 10 6 cells/ml) were cultured using a CellXVivo Human M2 Macrophage Differentiation Kit (R&D Systems) for 6 days in a humidified chamber with 5% CO 2 at 37°C.

Techniques:

Journal: Infection and Immunity

Article Title: Critical Role of the Interleukin-17/Interleukin-17 Receptor Axis in Regulating Host Susceptibility to Respiratory Infection with Chlamydia Species

doi: 10.1128/iai.00403-09

Figure Lengend Snippet: FIG. 3. Differential Th17-associated cytokine/chemokine responses induced in C3H/HeN and C57BL/6 mice following respiratory MoPn infection. C3H/HeN and C57BL/6 mice were sacrificed on days 6 and 12 after MoPn infection, and BAL fluid samples were collected as described in Materials and Methods. The amounts of IL-23 (A), IL-6 (B), CXCL1/KC (C), and CXCL2/MIP2 (D) in the BAL fluid samples were measured by ELISAs. The data are pooled from two independent experiments that showed similar trends and expressed as the means plus standard errors of the means (error bars) for 10 mice per time point per mouse strain. Values that were significantly different for the two strains by the unpaired, two-tailed Student t test are indicated by short lines and asterisks as follows: *, P 0.05; **, P 0.01.

Article Snippet: The cytokine contents in BAL fluids and culture supernatants were determined by using antibody pairs specific for mouse IFN- , IL-12, IL-17, IL-4, IL-6, IL-13, CXCL1/keratinocyte-derived chemokine (KC), and CXCL2/macrophage inflammatory protein 2 (MIP2) (all purchased from R&D Systems) and IL-23 (purchased from eBioscience).

Techniques: Infection, Two Tailed Test

Journal: Infection and Immunity

Article Title: Critical Role of the Interleukin-17/Interleukin-17 Receptor Axis in Regulating Host Susceptibility to Respiratory Infection with Chlamydia Species

doi: 10.1128/iai.00403-09

Figure Lengend Snippet: FIG. 4. Differential IL-17RC expression and differential IL-17 responsiveness of primary lung fibroblasts of naïve C3H/HeN and C57BL/6 mice. (A) Total RNA samples were prepared from the lungs or cultured primary fibroblasts for naïve C3H/HeN and C57BL/6, and the level of IL-17RA and IL-17RC expression was determined by quantitative RT-PCR and normalized against the housekeeping gene GAPDH (Rel, relative). Lung data are expressed as the means plus standard errors of the means (SEMs) (error bars) from five mice per strain. Fibroblast data were done in triplicate samples per strain. (B to D) Primary lung fibroblasts were isolated from C3H/HeN and C57BL/6 mice and stimulated with different concentrations of IL-17 or control medium for 24 h. The culture supernatants were used to measure the production of IL-6 (C), CXCL1/KC (B), and CXCL/MIP2 (D). The data are representative of two independent experiments, and the data are expressed as the means plus SEM s (error bars) of triplicate wells for each condition. Values that were significantly different in the two strains by the unpaired, two-tailed Student t test are indicated by short lines and asterisks as follows: *, P 0.05; **, P 0.01; ***, P 0.001.

Article Snippet: The cytokine contents in BAL fluids and culture supernatants were determined by using antibody pairs specific for mouse IFN- , IL-12, IL-17, IL-4, IL-6, IL-13, CXCL1/keratinocyte-derived chemokine (KC), and CXCL2/macrophage inflammatory protein 2 (MIP2) (all purchased from R&D Systems) and IL-23 (purchased from eBioscience).

Techniques: Expressing, Cell Culture, Quantitative RT-PCR, Isolation, Control, Two Tailed Test

Journal: Cells

Article Title: A Novel Treatment for Glomerular Disease: Targeting the Activated Macrophage Folate Receptor with a Trojan Horse Therapy in Rats

doi: 10.3390/cells10082113

Figure Lengend Snippet: Anti-inflammatory effect of EC2319. ( A ) Immunohistochemistry stained for ED1 + monocyte/macrophages, CD163 + , and CD 169 + infiltrates of kidney sections of rats with anti-GBM GN. ( B ) Quantification of ED1 + , CD163 + , and CD169 + infiltration. In rats treated with EC2319, the percentage of glomerular and interstitial M1 macrophages was significantly reduced without affecting the M2 macrophage phenotype. ( C , D ) RNase protection assay of cytokines and chemokines expressed in the glomeruli of anti-GBM glomerulonephritis in WKY rats. EC2319 decreases the expression of TGF-β and MCP-1/CCL2. ( E ). Spleen weight in control rats and EC2319 rats. Splenomegaly was prevented in the EC2319-treated group. At least sixty glomeruli per section were counted. For RPA, the data are presented as a ratio of the cpm for the specific mRNA/L-32 mRNA to ensure a constant quantity of RNA in each sample. Spleen weight is expressed as relative to body weight. Results were sampled from 6 control and 6 treated rats with anti-GBM GN and expressed as mean ± SEM. For RPA, 3 normal WKY rats were included. * p < 0.05, ** p < 0.01, *** p < 0.001. In ( B ): • Control, ■ EC2319. In ( C – E ): • Normal, ■ Control, ▲ EC2319.

Article Snippet: Infiltrating leukocytes were immunohistochemically stained for ED1 + (catalog MCA341R AbD Serotec), CD169 + , clone ED3, a marker for MI macrophages (catalog MCA343GA, AbD Serotec), and CD163 + , clone ED2, a marker for M2 macrophages (catalog MCA342R, AbD Serotec) infiltrates.

Techniques: Immunohistochemistry, Staining, Rnase Protection Assay, Expressing, Control

Journal: Journal of Tissue Engineering

Article Title: Repopulation of decellularised porcine pulmonary valves in the right ventricular outflow tract of sheep: Role of macrophages

doi: 10.1177/20417314221102680

Figure Lengend Snippet: Antibodies and methods used for detection of cells in pulmonary root tissues.

Article Snippet: CD163 M2- macrophages , AbD Serotec MCA 1853 IgG1; 1/100 , Proteinase K , DAKO Envision , Sheep lymph node , DAKO X0931 IgG1; 1/10 , NBF.

Techniques: Control

Journal: Asian Pacific Journal of Cancer Prevention : APJCP

Article Title: The Distribution of M2 Macrophage and Treg in Nasopharyngeal Carcinoma Tumor Tissue and the Correlation with TNM Status and Clinical Stage

doi: 10.31557/APJCP.2021.22.11.3447

Figure Lengend Snippet: M2 Macrophage (CD163) Density in Patients with NPC. Eight percent (8%; 4/50) of patients were negative result of CD163, 26% (13/50) were scant grade, 36% (18/50) were focal grade and 30% (15/50) were abundant grade

Article Snippet: Monoclonal anti-CD163 antibody (clone EPR19518, Abcam, Cambridge, UK) at 1:500 dilution in phosphate buffer saline (PBS) used to detect M2 macrophages and monoclonal anti-FoxP3 antibody (clone 236A/E7, Abcam, Cambridge, UK) at 1:100 dilution in PBS used to detect Treg.

Techniques:

Journal: Asian Pacific Journal of Cancer Prevention : APJCP

Article Title: The Distribution of M2 Macrophage and Treg in Nasopharyngeal Carcinoma Tumor Tissue and the Correlation with TNM Status and Clinical Stage

doi: 10.31557/APJCP.2021.22.11.3447

Figure Lengend Snippet: Immunohistochemistry of M2 Macrophage Using Anti-CD163 Antibody. Positive staining was shown on the membrane and cytoplasm (black arrow). The images were at 400x magnification. a. negative control (liver mouse) without anti-CD163 antibody, b. positive control (liver mouse) with anti-CD163 antibody, c. negative (0 score), no staining; d. scant (1 score), a small amount of scattered staining; e. focal (2 score), concentrated stains with an irregular and non-continuous focus; and f. abundant (3 score). Scale bar, 30 µm

Article Snippet: Monoclonal anti-CD163 antibody (clone EPR19518, Abcam, Cambridge, UK) at 1:500 dilution in phosphate buffer saline (PBS) used to detect M2 macrophages and monoclonal anti-FoxP3 antibody (clone 236A/E7, Abcam, Cambridge, UK) at 1:100 dilution in PBS used to detect Treg.

Techniques: Immunohistochemistry, Staining, Membrane, Negative Control, Positive Control

Journal: Asian Pacific Journal of Cancer Prevention : APJCP

Article Title: The Distribution of M2 Macrophage and Treg in Nasopharyngeal Carcinoma Tumor Tissue and the Correlation with TNM Status and Clinical Stage

doi: 10.31557/APJCP.2021.22.11.3447

Figure Lengend Snippet: The Correlation among the Clinical Parameters, M2 Macrophage and Tregs (n=50)

Article Snippet: Monoclonal anti-CD163 antibody (clone EPR19518, Abcam, Cambridge, UK) at 1:500 dilution in phosphate buffer saline (PBS) used to detect M2 macrophages and monoclonal anti-FoxP3 antibody (clone 236A/E7, Abcam, Cambridge, UK) at 1:100 dilution in PBS used to detect Treg.

Techniques: Significance Assay