Journal: Virology Journal

Article Title: Herpes simplex virus type 2 infection increases human immunodeficiency virus type 1 entry into human primary macrophages

doi: 10.1186/1743-422X-8-166

Figure Lengend Snippet: HSV-2 ability to infect human monocyte cell line U937 relies on their differentiation level . A) HSV-2 replication in undifferentiated U937 cells. The U937 cell line was infected with two different MOI of HSV-2, as reported in the graph legend. Vero cells were infected as a control. HSV-2 titre in the cellular supernatants was measured by plaque assay, at different times post-infection (p.i.). B) Effect of TPA treatment of U937 cells on CD14 expression. The effect of TPA treatment on U937 cell differentiation state was analyzed by CD14 FACS analysis. The percentage of positive cells is reported. C) HSV-2 replication in TPA-treated U937 cells. Undifferentiated or TPA-treated U937 cells were infected with HSV-2 (MOI of 1 PFU/cell). HSV-2 titre in the cellular supernatant was measured by plaque assay. In all cases, the reported values represent the mean of four independent experiments. The error bars represent the standard deviation.

Article Snippet: Briefly, 1 × 10 6 cells were harvested and directly incubated for one hour in cold PBS containing 1:100 (v/v) of an anti-human CD14 primary antibody (Li StarFISH).

Techniques: Infection, Control, Plaque Assay, Expressing, Cell Differentiation, Standard Deviation

Journal: Poultry Science

Article Title: Research note: Turkey ( Meleagris gallopavo ) semen CD14 + macrophages – management, key features, and classification – a pilot study with optimized protocols ☆

doi: 10.1016/j.psj.2025.105397

Figure Lengend Snippet: The visualization of turkey macrophages using immunofluorescence (IF) staining. A. A representative turkey semen CD14 + type 1 macrophages (CD14 + iNOS + , akin CD14 + Mф1); B. A representative turkey semen CD14 + type 2 macrophages (CD14 + IL10 + , akin CD14 + Mф2); C. A representative small mononucleated semen CD14 + Mф with an absence of vacuoles; d-F. Hypothetical process of multinucleated giant cell formation from two mononucleated small semen CD14 + Mф; G. Multinucleated giant semen CD14 + Mф with internalized sperm cells. Full arrows indicate vacuoles, and empty arrows indicate selected sperm. Images were captured using a confocal Laser Scanning Microscope (LSM800; Carl-Zeiss, Germany) equipped with an Airyscan super-resolution module (40 × /1.2 NA W c-apochromat) supported by Zen Blue Pro 2.6 software (Carl-Zeiss; Germany).

Article Snippet: Next, cells were triple-washed with PBS and covered with primary mouse anti-CD14 antibodies (orb434888, Biorbyt, United Kingdom, 1:50 in PBS).

Techniques: Immunofluorescence, Staining, Laser-Scanning Microscopy, Software

Journal: Poultry Science

Article Title: Research note: Turkey ( Meleagris gallopavo ) semen CD14 + macrophages – management, key features, and classification – a pilot study with optimized protocols ☆

doi: 10.1016/j.psj.2025.105397

Figure Lengend Snippet: A. Turkey semen CD14 + Mф measured by flow cytometry I-III: The representative cytogrammes show the gating strategy for identifying the different CD14 + Mф subpopulations. B. I-X: Consecutive photographs (10-min intervals) of turkey semen Mф phagocyting zymosan particle imaged using fluorescence microscopy with an Axio Observer. Z1/7 fluorescence microscope (Carl Zeiss, Inc., Oberkochen, Germany) equipped with ZEN 2.3 blue edition software (Carl Zeiss). C. Turkey semen Mф 24 h after seeding (magnification 200 ×), and D. after culture washing (magnification 400 ×). The high incidence of sperm in the cell pool decreases when cultured in high-glucose DMEM and after cell washing. Full arrows indicate selected Mф and empty arrows indicate sperm.

Article Snippet: Next, cells were triple-washed with PBS and covered with primary mouse anti-CD14 antibodies (orb434888, Biorbyt, United Kingdom, 1:50 in PBS).

Techniques: Flow Cytometry, Fluorescence, Microscopy, Software, Cell Culture

Journal:

Article Title: GM-CSF Regulates a PU.1-Dependent Transcriptional Program Determining the Pulmonary Response to LPS

doi: 10.1165/rcmb.2006-0174OC

Figure Lengend Snippet: GM-CSF, via PU.1, regulates mRNA levels for multiple components of the TLR-4 signaling pathway in AMs. mRNA transcripts encoding both positive and negative signaling components of the TLR-4 signaling pathway in cultured alveolar macrophages were detected by RT-PCR. Note the absence of transcripts for CD14, RP105, and IRAK-M in mAM cells.

Article Snippet: Briefly, primary antibodies included rat anti-murine CD14 (PharMingen, San Diego, CA) diluted 1:200, rabbit anti-mouse IRAK-M (Chemicon International, San Diego, CA) diluted 1:1,000, or goat anti-murine β-actin antibody (Santa Cruz, Santa Cruz, CA) diluted 1:200.

Techniques: Cell Culture, Reverse Transcription Polymerase Chain Reaction

Journal:

Article Title: GM-CSF Regulates a PU.1-Dependent Transcriptional Program Determining the Pulmonary Response to LPS

doi: 10.1165/rcmb.2006-0174OC

Figure Lengend Snippet: GM-CSF, via PU.1, regulates multiple components of TLR-4 signaling pathway in AMs. (A) Evaluation of RP105 on cultured alveolar macrophage cell lines by flow cytometry. Cells were immunostained with PE-labeled, anti-RP105 (black line) or isotype control (gray shading) antibodies and evaluated by flow cytometry. (B) RP105 levels on AMs were quantified by determining the difference between the mean fluorescence of respective cell lines immunostained with anti-RP105 antibody minus the isotype control antibody. RP105 was readily detectible on MH-S and mAMPU.1+ cells and absent on mAM cells (n = 3 determinations/cell line; P < 0.001 [Kruskal-Wallis ANOVA on ranks]). (C) Evaluation of CD14 and IRAK-M on cultured alveolar macrophage cell lines by Western blot analysis.

Article Snippet: Briefly, primary antibodies included rat anti-murine CD14 (PharMingen, San Diego, CA) diluted 1:200, rabbit anti-mouse IRAK-M (Chemicon International, San Diego, CA) diluted 1:1,000, or goat anti-murine β-actin antibody (Santa Cruz, Santa Cruz, CA) diluted 1:200.

Techniques: Cell Culture, Flow Cytometry, Labeling, Fluorescence, Western Blot

Journal: PeerJ

Article Title: Main histological parameters to be evaluated in an experimental model of myocardial infarct treated by stem cells on pigs

doi: 10.7717/peerj.7160

Figure Lengend Snippet: The number of macrophages (arrow) in the peri-cicatricial area of MI+ASC heart (A) is similar to MIC group (B). Immunohistochemistry anti-CD14, PAP. 400 ×. M2 macrophages (arrows) are in greater proportion in MI+ASC group (C) than in MIC (D). Immunohistochemistry anti-CD163 PAP. 400 ×.

Article Snippet: The primary antibodies used were anti-CD14 antibody (MyBioSource, MBS-2027456, 1/500), anti-CD163 antibody (Serotec, MCA242GA, 1/200), muscle specific actin monoclonal antibody (Novocastra, A7811, 1/100), Desmin Monoclonal Antibody (Novocastra, DE-R11, 1/50), Connexin 43 mouse monoclonal antibody (Cell Signaling Technology, cst-3512, 1/50), and HIF1-α antibody (Gene Tex, GTX 30105, 1/1000).

Techniques: Immunohistochemistry

Journal: PLoS ONE

Article Title: Glatiramer Acetate Increases Phagocytic Activity of Human Monocytes In Vitro and in Multiple Sclerosis Patients

doi: 10.1371/journal.pone.0051867

Figure Lengend Snippet: A–D. Immunocytochemistry showed distinct phagocytosis of polystyrene beads (green) by monocytes treated with 31.25 µg/ml glatiramer acetate (GA) for 24 h (red, labelled with anti-CD14 antibody) as compared to medium, human serum albumin (HA), and mannitol (Mt) vehicle control. Scale bar: 50 µm. E. Representative z-stack series of confocal microscopic images demonstrate that the particles were completely internalised and not merely attached to the outer membrane. Scale bar: 10 µm.

Article Snippet: After 30 minutes incubation with fluorescent latex beads (FluoresbriteTM YG carboxylate microspheres; 1 μm diameter; Polysciences, Eppelheim, Germany) at 37°C, cover slips were washed three times and cells were stained with the CD14 antibody (primary: rabbit anti-human IgG, Abcam, Cambridge, UK; secondary: goat anti-rabbit IgG, AbD Serotec, Düsseldorf, Germany) at 1∶100 dilution in DMEM+.

Techniques: Immunocytochemistry, Control, Membrane

Journal: PLoS ONE

Article Title: Glatiramer Acetate Increases Phagocytic Activity of Human Monocytes In Vitro and in Multiple Sclerosis Patients

doi: 10.1371/journal.pone.0051867

Figure Lengend Snippet: A–B. Gated monocytes in a forward vs. side scatter dot-plot analysis of peripheral blood mononuclear cells. Monocytes were then gated according to their surface expression of CD14 and CD16. Flow cytometric analysis of phagocytosis revealed that CD14 ++ CD16 + monocytes (gate III) engulfed polystyrene beads more effectively than the other subsets (gate I and II). CI–III. The mean fluorescence intensities represent the amount of incorporated fluorescent latex particles phagocytosed by 3×10 5 cells. D. Increase in the percentage of CD14 + CD16 + monocytes after 4 h and 8 h of treatment with 31.25 µg/ml glatiramer acetate (GA) in MACS isolated monocytes. Data are expressed as mean percentages ± SEM of three independent experiments. Significant effects vs. controls are indicated by asterisks (*p<0.05, **p<0.01, and ***p<0.001 using Bonferroni's Multiple Comparison Test) as determined by one-way ANOVA. E. Slight but not significant increase of CD16 expression after GA treatment.

Article Snippet: After 30 minutes incubation with fluorescent latex beads (FluoresbriteTM YG carboxylate microspheres; 1 μm diameter; Polysciences, Eppelheim, Germany) at 37°C, cover slips were washed three times and cells were stained with the CD14 antibody (primary: rabbit anti-human IgG, Abcam, Cambridge, UK; secondary: goat anti-rabbit IgG, AbD Serotec, Düsseldorf, Germany) at 1∶100 dilution in DMEM+.

Techniques: Expressing, Fluorescence, Isolation, Comparison

Journal: PLoS ONE

Article Title: Glatiramer Acetate Increases Phagocytic Activity of Human Monocytes In Vitro and in Multiple Sclerosis Patients

doi: 10.1371/journal.pone.0051867

Figure Lengend Snippet: Mean fluorescence intensities of CD36, CD210, and TIM-3 (± SEM) of three independent experiments after a treatment period of 24 h according to the different monocyte subsets, i.e. CD14 + CD16 − and CD14 + CD16 + monocytes.

Article Snippet: After 30 minutes incubation with fluorescent latex beads (FluoresbriteTM YG carboxylate microspheres; 1 μm diameter; Polysciences, Eppelheim, Germany) at 37°C, cover slips were washed three times and cells were stained with the CD14 antibody (primary: rabbit anti-human IgG, Abcam, Cambridge, UK; secondary: goat anti-rabbit IgG, AbD Serotec, Düsseldorf, Germany) at 1∶100 dilution in DMEM+.

Techniques: Fluorescence

Journal: PLoS ONE

Article Title: Glatiramer Acetate Increases Phagocytic Activity of Human Monocytes In Vitro and in Multiple Sclerosis Patients

doi: 10.1371/journal.pone.0051867

Figure Lengend Snippet: The MFI displays the amount of incorporated beads phagocytosed by 3×10 5 cells. Data are expressed as means of the mean fluorescence intensities (MFI) ± SEM of three independent experiments. Pre-treatment of monocytes with anti-CD14, anti-CD16, anti-CD32, anti-CD210, and anti-TIM3 antibodies reduced glatiramer acetate (GA) induced phagocytosis of polystyrene beads. The anti-CD36 and, at the highest concentration, the anti-IL10 antibody most effectively suppressed phagocytosis. Significant effects vs. GA 31.25 µg/ml are indicated by asterisks (*p<0.05, **p<0.01, and ***p<0.001 using Bonferroni's Multiple Comparison Test) as determined by one-way ANOVA. B–C. The anti-CD36 antibodies of two different clones show a diverse impact on the phagocytosis of polystyrene beads by untreated cells, whereas both antibodies effectively decreased the GA induced phagocytosis.

Article Snippet: After 30 minutes incubation with fluorescent latex beads (FluoresbriteTM YG carboxylate microspheres; 1 μm diameter; Polysciences, Eppelheim, Germany) at 37°C, cover slips were washed three times and cells were stained with the CD14 antibody (primary: rabbit anti-human IgG, Abcam, Cambridge, UK; secondary: goat anti-rabbit IgG, AbD Serotec, Düsseldorf, Germany) at 1∶100 dilution in DMEM+.

Techniques: Fluorescence, Concentration Assay, Comparison, Clone Assay

Journal: ERJ Open Research

Article Title: The effects of exogenous lipid on THP-1 cells: an in vitro model of airway aspiration?

doi: 10.1183/23120541.00026-2016

Figure Lengend Snippet: PMA/THP-1 lipid uptake. PMA/THP-1 cells were incubated with 10% vol/vol Calogen for 24 h with and without inhibitors and were stained with ORO to detect lipid accumulation. a) Untreated cells were compared with b) cells treated with Calogen alone and c) in the presence of cytochalasin D and d) CD36 monoclonal blocking antibody. Inset images are magnified representative regions of each treatment. e) Cells treated with Calogen were also visualised under TEM. Scale bar=1 μm. f, g) Lipid droplets were identified and magnified to reveal dark stained remnants of lipid encased in membrane-bound vesicles, as indicated by the arrows. Scale bars=500 nm (f) and 100 nm (g).

Article Snippet: The plate was centrifuged (350 g , 3 min, 4 °C) and washed twice with PBS before incubating (30 min at 4 °C) with mouse monoclonal primary antibodies (CD11b, CD14, CD16, CD23, CD24, CD32, CD36, CD64, CD80, CD86, CD163, CD206 (20 μg·mL −1 , AbD Serotec, Oxford, UK) or CD32 [ ]).

Techniques: Incubation, Staining, Blocking Assay, Membrane