Journal: Cancers

Article Title: Murlentamab, a Low Fucosylated Anti-Müllerian Hormone Type II Receptor (AMHRII) Antibody, Exhibits Anti-Tumor Activity through Tumor-Associated Macrophage Reprogrammation and T Cell Activation

doi: 10.3390/cancers13081845

Figure Lengend Snippet: Flow cytometry antibodies used.

Article Snippet: CD36-PEVio770 , Miltenyi , 130-110-742 , REA770 , .

Techniques: Flow Cytometry, In Vivo, In Vitro

Journal: Cancers

Article Title: Murlentamab, a Low Fucosylated Anti-Müllerian Hormone Type II Receptor (AMHRII) Antibody, Exhibits Anti-Tumor Activity through Tumor-Associated Macrophage Reprogrammation and T Cell Activation

doi: 10.3390/cancers13081845

Figure Lengend Snippet: Murlentamab opsonization of SKOV3-R2 + orients naïve macrophages and reprograms TAMs towards an M1-like profile. SKOV3-R2 + ovarian tumor cells were labeled with different 3C23K antibodies (3C23K-FcKO control, 3C23K-CHO normally fucosylated or murlentamab the low fucosylated form) and cultured in the presence of human monocyte-derived macrophages from healthy donors unstimulated (M0) or stimulated with M-CSF and IL-10 (TAMs). ( A ) The proportion of macrophages expressing M1/M2 membrane markers (CD32, CD64, CD80, TLR2, CD163, CD36 and CD206) was determined by flow cytometry after three days of co-culture with SKOV3-R2 + cells. ( B ) The release of cytokines (IL1β, IL12, TNFα, IL6, IFNγ, IL10) and chemokines (CCL2, CCL4, CCL5, CXCL9 and CXCL10) in the culture medium was determined by AlphaLISA after three days of co-culture with SKOV3-R2 + cells. Data shown (boxplots) are the results from three different experiments (performed with three different healthy donors). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. p values were determined using one-way ANOVA analysis followed by Tukey’s multiple comparisons test.

Article Snippet: CD36-PEVio770 , Miltenyi , 130-110-742 , REA770 , .

Techniques: Labeling, Cell Culture, Derivative Assay, Expressing, Membrane, Flow Cytometry, Co-Culture Assay

Journal: Cell

Article Title: A blood atlas of COVID-19 defines hallmarks of disease severity and specificity

doi: 10.1016/j.cell.2022.01.012

Figure Lengend Snippet:

Article Snippet: CD36 (5-271)-155Gd , Fluidigm , Cat# 3155012B, RRID: AB_2756286.

Techniques: Mass Cytometry, Flow Cytometry, Recombinant, Staining, Selection, Antibody Labeling, Labeling, Isolation, Sample Prep, Luminex, Quantitative Proteomics, Generated, Gene Expression, Clone Assay, Marker, Expressing, Mass Spectrometry, Derivative Assay, RNA Sequencing, Sequencing, Illumina Sequencing, Software, Variant Assay

Journal: Respiratory Research

Article Title: An investigation of the resolution of inflammation (catabasis) in COPD

doi: 10.1186/1465-9921-13-101

Figure Lengend Snippet: Representative examples of different macrophage immunostaining intensities (absent/moderate vs. intense staining) for CD44, CD36, VEGF and TGFβ. For further explanations, see text.

Article Snippet: Formalin-fixed paraffin-embebbed tissue sections (3 μm thick) were immunostained with the following monoclonal mouse antibodies: anti-human CD44, Phagocytic Glycoprotein-1, clone DF1485 (Dako, Glostrup, Denmark); anti-human VEGF, clone VG1 (Dako, Glostrup, Denmark); anti-human TGFbeta (AbDSerotec, Oxford, UK); anti-human CD36, clone SMO, (AbDSerotec, Oxford, UK).

Techniques: Immunostaining, Staining

Journal: Respiratory Research

Article Title: An investigation of the resolution of inflammation (catabasis) in COPD

doi: 10.1186/1465-9921-13-101

Figure Lengend Snippet: Individual and mean (bars) values of the proportion of macrophages with intense staining for CD44 , CD36, VEGF and TGFβ in patients with COPD, smokers with normal spirometry and non-smokers. (S = current smokers; EX-S = former smokers; NS = non-smokers). For further explanations, see text.

Article Snippet: Formalin-fixed paraffin-embebbed tissue sections (3 μm thick) were immunostained with the following monoclonal mouse antibodies: anti-human CD44, Phagocytic Glycoprotein-1, clone DF1485 (Dako, Glostrup, Denmark); anti-human VEGF, clone VG1 (Dako, Glostrup, Denmark); anti-human TGFbeta (AbDSerotec, Oxford, UK); anti-human CD36, clone SMO, (AbDSerotec, Oxford, UK).

Techniques: Staining

Journal: Scientific Reports

Article Title: Endothelial Cell Phenotypes are Maintained During Angiogenesis in Cultured Microvascular Networks

doi: 10.1038/s41598-018-24081-z

Figure Lengend Snippet: Illustration of CD36 labeling in ex vivo cultured microvascular networks and stimulated in vivo microvasculature. Comparison between Day 3 ( Ex Vivo ) ( A–C ) and Day 3 ( In Vivo ) ( D–F ) microvascular networks revealed diminishing CD36 labeling along the length of the capillary sprout becoming absent at the tip. Arrows identify examples of CD36-negative capillary sprouts. Scale bars = 200 µm. ( G , H ) Capillary sprouts exhibited a substantial decrease in CD36 labeling compared to larger network vessels ( > 10 µm) in Day 3 ( Ex Vivo ) and Day 3 ( In Vivo ) tissues. White and black bars represent “Sprouts” and “Networks” respectively for Day 3 ( Ex Vivo ) and Day 3 ( In Vivo ) groups. The *** indicates a significant difference of p < 0.001 by Mann-Whitney U test.

Article Snippet: The following primary antibodies were used: UNC5b (1:100; Abcam; Cambridge, MA), VEGFR-2 (1:50; Santa Cruz Biotechnology; Dallas, TX), Alexa-568 Phalloidin (1:50; Invitrogen; Carlsbad, CA), CD36 (1:100; Novus Biologicals; Littleton, CO), and PECAM (1:200; BD Pharmingen; San Jose, CA).

Techniques: Labeling, Ex Vivo, Cell Culture, In Vivo, Comparison, MANN-WHITNEY

Journal: The Journal of Immunology Author Choice

Article Title: CD36-Mediated Hematoma Absorption following Intracerebral Hemorrhage: Negative Regulation by TLR4 Signaling

doi: 10.4049/jimmunol.1400054

Figure Lengend Snippet: Reduced hematoma absorption and aggravated neurologic deficits in CD36-deficient patients with ICH. (A) Typical cranial CT images of CD36-normal (n = 11) and CD36-deficient (n = 11) ICH patients. Original magnification ×10. (B) Comparison of hematoma volumes in the CD36-deficient and CD36-normal groups at admission and 7 d after the onset of ICH. *p < 0.05 versus the CD36-normal group. (C) Decreased absorption rate in the CD36-deficient patients relative to the normal group. **p < 0.01 versus the CD36-normal group. (D) NIHSS scores and (E) mRS scores for ICH patients. *p < 0.05 versus the CD36-normal group, **p < 0.01 versus the CD36-normal group.

Article Snippet: The membranes were incubated with mouse anti-human CD36 (for human blood, 1:400; Wako Chemicals USA, Richmond, VA) or rabbit anti-mouse CD36 (for perihematomal tissues and cells, 1:400; Abcam) at 4°C overnight; GAPDH (1:400; Santa Cruz Biotechnology, Dallas, TX) was used as a loading control.

Techniques:

Journal: The Journal of Immunology Author Choice

Article Title: CD36-Mediated Hematoma Absorption following Intracerebral Hemorrhage: Negative Regulation by TLR4 Signaling

doi: 10.4049/jimmunol.1400054

Figure Lengend Snippet: Enhanced CD36 expression in perihematomas following ICH. (A) Detection of CD36 mRNA expression in mice subjected to ICH using real-time quantitative RT-PCR. The data are expressed as fold increases relative to naive animals. **p < 0.01 versus sham, n = 6. (B) Detection of CD36 protein expression in mice subjected to ICH using Western blot. **p < 0.01 versus sham, n = 6. (C) Detection of CD36 expression in astrocytes, neurons, and microglia in the perihematomal tissues of mice using flow cytometry at 3 d after ICH. The left panel is the representative flow cytometry plot and the lower right panel is the percentage of CD36+ cells in neurons, astrocytes, and microglia. Microglia were marked by CD45intCD11b+, whereas neurons were identified as β-III tubulin+, and astrocytes were identified as GFAP+. **p < 0.01 versus sham, ##p < 0.01 versus astrocytes or neurons, n = 3. (D) Detection of CD36 expression in human perihematomal tissues of ICH patients using fluorescence immunohistochemistry; CD36 expression was labeled with a CD36 Ab (red), astrocytes, neurons, and microglia were labeled with GFAP (green), Neun (green), and Iba-1 (green), respectively. The merged images of the overlay of CD36 together with astrocytes, neurons, and microglia were shown as yellow, and the nuclei were stained with DAPI (blue). The arrows indicate positive cells. Scale bars, 80 μm.

Article Snippet: The membranes were incubated with mouse anti-human CD36 (for human blood, 1:400; Wako Chemicals USA, Richmond, VA) or rabbit anti-mouse CD36 (for perihematomal tissues and cells, 1:400; Abcam) at 4°C overnight; GAPDH (1:400; Santa Cruz Biotechnology, Dallas, TX) was used as a loading control.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Flow Cytometry, Fluorescence, Immunohistochemistry, Labeling, Staining

Journal: The Journal of Immunology Author Choice

Article Title: CD36-Mediated Hematoma Absorption following Intracerebral Hemorrhage: Negative Regulation by TLR4 Signaling

doi: 10.4049/jimmunol.1400054

Figure Lengend Snippet: CD36-mediated hematoma absorption in mice with ICH. (A) Serial coronal sections of mouse brain tissues 5 d after the onset of ICH. Original magnification ×1.5. (B) Comparison of hematoma volumes between WT and CD36−/− mice. The data in the left and right panels are the Image-Pro Plus and hemoglobin detection results, respectively. **p < 0.01 versus the WT group, n = 6. (C) The NDS of WT and CD36−/− mice at 1, 3, 5, and 7 d after the onset of ICH. *p < 0.05 versus the WT group at the corresponding time points, n = 6. (D) Brain water content and (E) mRNA expression of inflammatory factors 3 d after the onset of ICH. The data in (E) were assessed by real-time quantitative RT-PCR and were expressed as fold increases relative to naive animals. *p < 0.05 versus the WT group; **p < 0.01 versus the WT group, n = 6. Cereb, cerebellum; Cont BG, contralateral basal ganglia; Cont CX, contralateral cortex; Ipsi BG, ipsilateral basal ganglia; Ipsi CX, ipsilateral cortex.

Article Snippet: The membranes were incubated with mouse anti-human CD36 (for human blood, 1:400; Wako Chemicals USA, Richmond, VA) or rabbit anti-mouse CD36 (for perihematomal tissues and cells, 1:400; Abcam) at 4°C overnight; GAPDH (1:400; Santa Cruz Biotechnology, Dallas, TX) was used as a loading control.

Techniques: Expressing, Quantitative RT-PCR

Journal: The Journal of Immunology Author Choice

Article Title: CD36-Mediated Hematoma Absorption following Intracerebral Hemorrhage: Negative Regulation by TLR4 Signaling

doi: 10.4049/jimmunol.1400054

Figure Lengend Snippet: CD36-mediated engulfment of RBCs by microglia. (A) Observation of the phagocytosis of RBCs (CFSE-labeled, green) by microglia (Alexa Fluor 594–labeled, red) using confocal microscopy. The nuclei were stained with Hoechst. Serial sections along the z-axis were acquired and compiled as images. The x–z-axis is below, and the y–z-axis is on the right. The arrows indicate the RBCs engulfed by microglia. Scale bars, 40 μm. Detection of changes in the phagocytic ability of microglia using flow cytometry is shown. Microglia were stained with anti-mouse CD11b-PE and RBCs were stained with CFSE. (B) Representative flow cytometry plot (left panel) and the percentage of PE+CFSE+ microglia that had engulfed RBCs (right panel). (C) Average fluorescence intensity of CFSE in the PE+CFSE+ microglia. (D) The number of nonphagocytosed RBCs in the supernatant. **p < 0.01 versus the WT microglia group, n = 6 for all graphs.

Article Snippet: The membranes were incubated with mouse anti-human CD36 (for human blood, 1:400; Wako Chemicals USA, Richmond, VA) or rabbit anti-mouse CD36 (for perihematomal tissues and cells, 1:400; Abcam) at 4°C overnight; GAPDH (1:400; Santa Cruz Biotechnology, Dallas, TX) was used as a loading control.

Techniques: Labeling, Confocal Microscopy, Staining, Flow Cytometry, Fluorescence

Journal: The Journal of Immunology Author Choice

Article Title: CD36-Mediated Hematoma Absorption following Intracerebral Hemorrhage: Negative Regulation by TLR4 Signaling

doi: 10.4049/jimmunol.1400054

Figure Lengend Snippet: TLR4 signaling downregulates CD36 expression and increases hematoma absorption. (A) Detection of CD36 expression in perihematomal tissues 3 d after the onset of ICH using Western blot. **p < 0.01 versus the WT group, n = 6. (B) Detection of microglia CD36 expression using flow cytometry. **p < 0.01 versus the WT group, n = 6. (C) Five days after the onset of ICH, the hematoma volume was smaller in MyD88−/− and TLR4−/− mice than in WT mice. Measurement of hematoma volume using Image-Pro Plus (left panel) and hemoglobin detection (right panel) are shown. **p < 0.01 versus the WT group, n = 6. Detection of changes in the phagocytic capacity of microglia using flow cytometry is depicted. Microglia were stained with anti-mouse CD11b-PE, and RBCs were stained with CFSE. (D) Representative flow cytometry plot (left panel) and the percentage of PE+CFSE+ microglia that had engulfed RBCs (right panel); (E) average fluorescence intensity of CFSE in PE+CFSE+ microglia; (F) number of nonphagocytosed RBCs in the supernatant. **p < 0.01 versus the WT microglia group, ##p < 0.01 versus the groups with anti-CD36 Abs, n = 6 for all flow cytometry graphs.

Article Snippet: The membranes were incubated with mouse anti-human CD36 (for human blood, 1:400; Wako Chemicals USA, Richmond, VA) or rabbit anti-mouse CD36 (for perihematomal tissues and cells, 1:400; Abcam) at 4°C overnight; GAPDH (1:400; Santa Cruz Biotechnology, Dallas, TX) was used as a loading control.

Techniques: Expressing, Western Blot, Flow Cytometry, Staining, Fluorescence

Journal: The Journal of Immunology Author Choice

Article Title: CD36-Mediated Hematoma Absorption following Intracerebral Hemorrhage: Negative Regulation by TLR4 Signaling

doi: 10.4049/jimmunol.1400054

Figure Lengend Snippet: Effects of TNF-α, IL-1β, and IL-10 on CD36 expression and phagocytosis in microglia. (A) Detection of changes in microglial CD36 protein levels using Western blot. *p < 0.05 versus the vehicle group, **p < 0.01 versus the vehicle group, n = 6. (B) Detection of microglial CD36 expression using flow cytometry. **p < 0.01 versus the vehicle group, n = 6. Detection of changes in the phagocytic ability of microglia using flow cytometry. Microglia were stained with anti-mouse CD11b-PE, and RBCs were stained with CFSE. (C) Representative flow cytometry plot (left panel) and the percentage of PE+CFSE+ microglia that had engulfed RBCs (right panel). **p < 0.01 versus the vehicle group, n = 6. (D) Average fluorescence intensity of CFSE in PE+CFSE+ microglia. **p < 0.01 versus the vehicle group, n = 6. (E) Number of nonphagocytosed RBCs in the supernatant. *p < 0.05 versus the vehicle group, **p < 0.01 versus the vehicle group, n = 6.

Article Snippet: The membranes were incubated with mouse anti-human CD36 (for human blood, 1:400; Wako Chemicals USA, Richmond, VA) or rabbit anti-mouse CD36 (for perihematomal tissues and cells, 1:400; Abcam) at 4°C overnight; GAPDH (1:400; Santa Cruz Biotechnology, Dallas, TX) was used as a loading control.

Techniques: Expressing, Western Blot, Flow Cytometry, Staining, Fluorescence

Journal: The Journal of Immunology Author Choice

Article Title: CD36-Mediated Hematoma Absorption following Intracerebral Hemorrhage: Negative Regulation by TLR4 Signaling

doi: 10.4049/jimmunol.1400054

Figure Lengend Snippet: TAK-242 (TAK) increases CD36 expression and promotes hematoma absorption. (A) Detection of CD36 protein expression using Western blot. **p < 0.01 versus the vehicle group, #p < 0.05 versus the TAK group, ##p < 0.01 versus the TAK group, n = 6. (B) Detection of microglia CD36 expression using flow cytometry. **p < 0.01 versus the vehicle group, ##p < 0.01 versus the TAK group, n = 6. Detection of changes in the phagocytic capacity of microglia using flow cytometry. Microglia were stained with anti-mouse CD11b-PE, and RBCs were stained with CFSE. (C) A representative flow cytometry plot (left panel) displaying the percentage of PE+CFSE+ microglia (right panel) that had engulfed RBCs. **p < 0.01 versus the vehicle group, ##p < 0.01 versus the TAK group, n = 6. (D) The average fluorescence intensity of CFSE in PE+CFSE+ microglia. **p < 0.01 versus the vehicle group, ##p < 0.01 versus the TAK group, n = 6. (E) The number of nonphagocytosed RBCs in the supernatant. **p < 0.01 versus the vehicle group, ##p < 0.01 versus the TAK group, n = 6. (F) Detection of microglial catalase expression using real-time quantitative RT-PCR. The data are expressed as fold increases relative to the vehicle group. **p < 0.01, n = 6. (G) The changes in H2O2 content in the supernatant were measured. The results are presented as fold changes relative to vehicle group. **p < 0.01, n = 6. (H) Changes in hematoma volume in mice at 5 d after the onset of ICH. Measurement of hematoma volume using Image-Pro Plus (left panel) and detection of hemoglobin (right panel). **p < 0.01 versus the ICH plus vehicle group, #p < 0.05 versus the ICH plus TAK plus TNF-α group, n = 6.

Article Snippet: The membranes were incubated with mouse anti-human CD36 (for human blood, 1:400; Wako Chemicals USA, Richmond, VA) or rabbit anti-mouse CD36 (for perihematomal tissues and cells, 1:400; Abcam) at 4°C overnight; GAPDH (1:400; Santa Cruz Biotechnology, Dallas, TX) was used as a loading control.

Techniques: Expressing, Western Blot, Flow Cytometry, Staining, Fluorescence, Quantitative RT-PCR