Journal: Malaria Journal

Article Title: Real-time measurement of Plasmodium falciparum -infected erythrocyte cytoadhesion with a quartz crystal microbalance

doi: 10.1186/s12936-016-1374-7

Figure Lengend Snippet: Quartz coating with CD36 using membranes of melanoma cells and CSA via PLL. a Overlay picture of a C32 melanoma cell stained for DNA (DAPI, blue ) and CD36 (FITC, green ) showing a homogenous spread of CD36 on the surface and the nucleus in the centre of the cell; b fluorescence picture of isolated and homogenized cell membranes of C32 melanoma cells immobilized to a PLL-coated quartz and stained with a CD36 antibody (FITC, green ); c overlay picture of a quartz coated with PLL/CSA showing an even distribution of fluorescence signal, indicating a homogenous coating of CSA to the quartz. The quartz was stained with an anti-CSA antibody (FITC, green )

Article Snippet: C32-melanoma cells expressing large amounts of CD36 were obtained from the American Type Culture Collection (ATCC) and cultured at 5 % carbon dioxide and 37 °C using DMEM supplemented with 50 μg/ml gentamycin, 10 % FCS, 2 mM l -glutamine, and 1 % MEM non-essential amino acid solution (NEAA).

Techniques: Staining, Fluorescence, Isolation

Journal: Malaria Journal

Article Title: Real-time measurement of Plasmodium falciparum -infected erythrocyte cytoadhesion with a quartz crystal microbalance

doi: 10.1186/s12936-016-1374-7

Figure Lengend Snippet: Overview of QCM experiments measuring attachment of cells to receptors by frequency shifts and subsequent microscopic count. Median, interquartile range and single measurements of frequency shifts measured by the QCM platform F id g et Type 1 ( upper panel ) and subsequent microscopic count of the number of attached cells per microscopic field ( lower panel ) for either P. falciparum iRBCs of FCR3-CSA strain (n = 5) and red blood cells (RBC, n = 3) added to CSA receptors (CSA), or for iRBCs of FCR3-CD36 strain (n = 6) and RBCs (n = 4) added to CD36 receptors. Results show that iRBCs (FCR3 parasites) bind to the respective receptor, reflected by a dampening of the frequency and a higher count data in the microscope whereas RBCs do not bind. CSA chondroitin sulfate A, PLL poly- l -lysin

Article Snippet: C32-melanoma cells expressing large amounts of CD36 were obtained from the American Type Culture Collection (ATCC) and cultured at 5 % carbon dioxide and 37 °C using DMEM supplemented with 50 μg/ml gentamycin, 10 % FCS, 2 mM l -glutamine, and 1 % MEM non-essential amino acid solution (NEAA).

Techniques: Microscopy

Journal: Malaria Journal

Article Title: Real-time measurement of Plasmodium falciparum -infected erythrocyte cytoadhesion with a quartz crystal microbalance

doi: 10.1186/s12936-016-1374-7

Figure Lengend Snippet: Example QCM signals and pictures obtained for CD36. Measurements were carried out on the two-channel sensor platform F id g et Type 1. a The samples (iRBCs and RBCs) were injected after achievement of a stable baseline followed by a short stop-flow interval. Adhesion of iRBCs (FCR3-CD36) led to a decrease of the signal. The low frequency shift obtained with RBCs shows an unspecific drift due to the alterations in the viscosity of the fluid. Frequency shifts after 2.5 h (time frame indicated by vertical dotted lines ) are −83 Hz for RBCs and −361 Hz for iRBCs indicating the attachment of iRBCs to CD36 on the quartz; b and c pictures of corresponding quartzes stained with DAPI after measurement in the QCM platform confirming the results. After injection of iRBCs it can be clearly seen in ( b ) that iRBCs bind to the attached C32 cell membranes expressing CD36 on the quartz. In contrast in ( c ), after the experiment with RBCs only very few RBCs are detected on the sensor surface and only cell membranes can be seen

Article Snippet: C32-melanoma cells expressing large amounts of CD36 were obtained from the American Type Culture Collection (ATCC) and cultured at 5 % carbon dioxide and 37 °C using DMEM supplemented with 50 μg/ml gentamycin, 10 % FCS, 2 mM l -glutamine, and 1 % MEM non-essential amino acid solution (NEAA).

Techniques: Injection, Viscosity, Staining, Expressing

Journal: Cancers

Article Title: MCF-7 Drug Resistant Cell Lines Switch Their Lipid Metabolism to Triple Negative Breast Cancer Signature

doi: 10.3390/cancers13235871

Figure Lengend Snippet: BCC CM enhances FABP4, FABP5 and CD36 mRNA levels and increases their releasement to media. ( A )Graph representation of FABP4, FABP5 and CD36 transcripts in mature adipocytes at different conditions for 24 and 48 h of treatment. There is a clear and significant increase of all three transcripts in those mature adipocytes that have been culture with BCC line CM. Interestingly, FABP4 transcript in mature adipocytes culture with MCF-7 CM is not affected unlike the rest of conditions and transcripts. One-way ANOVA where a p < 0.05 was considered statistically significant. ( B ) Media from control adipocytes is compared with media from adipocytes cultured with MDA-MB-231 and MCF-7 cell lines conditioned media, for 24 h where a p -value < 0.05 is statistically significant (* < 0.05, ** < 0.005, *** < 0.0005, **** < 0.0001).

Article Snippet: Levels of mRNA were assessed using LightCycler96 device (Roche) with the Taqman probes for respective genes acquired from Life technologies CD36 (Mm00432403_m1), FABP4 (Mm00445878_m1), FABP5 (Mm00783731_s1), TBP (Mm01277042_m1).

Techniques: Control, Cell Culture

Journal: PLoS ONE

Article Title: Oxidation by Neutrophils-Derived HOCl Increases Immunogenicity of Proteins by Converting Them into Ligands of Several Endocytic Receptors Involved in Antigen Uptake by Dendritic Cells and Macrophages

doi: 10.1371/journal.pone.0123293

Figure Lengend Snippet: ( A ) Uptake of fluorescently-labelled proteins by CHO cells transfected with human CD36 as compared to non-transfected cells. ( B ) Binding of anti-CD36 mAb and control mouse IgA to CD36-transfected and non-transfected CHO cells, determined by cellular ELISA. ( C ) Binding of rCD36 to plate-adsorbed proteins. ( D ) Effects of 10 or 100 μg/ml of indicated, soluble ligands on rCD36 binding to plate-adsorbed OVA-Cl. ( E ) Binding of polyclonal anti-mouse CD36 Ab and control goat IgG to BM-DC, splenic DC and PEM. A representative histogram of Ab binding to BM-DC is displayed on the left graph. ( F ) Binding of anti-mouse SR-A mAb and control rat IgG2b to BM-DC, splenic DC and PEM. A representative histogram of Ab binding to BM-DC is shown on the left graph. ( G ) Binding of SR-A present in lysates of PEM to plate-adsorbed proteins. ( H ) Effects of anti-SR-A 2F8 mAb and AcLDL on the uptake of fluorescently-labelled proteins by BM-DC. Shown are results of single experiments, each representative of at least 3 similar experiments performed (A-D, G, H) or averages +SEM from 4–6 independent experiments (E, F). The data were analysed with the unpaired (A-C, G) or one-sample (H) Student’s t-test or with ANOVA, followed by the Tukey-Kramer post-test (D). *, p < 0.05; ND, not done.

Article Snippet: Rat anti-mouse scavenger receptor A (SR-A) mAb (clone 2F8) was obtained from AbD Serotec; mouse anti-mouse CD36 mAb (CRF D-2712) from Hycult Biotech; mouse IgA isotype control mAb (M18-254), rat IgG2b isotype control mAb (A95-1), rat anti-mouse CD11b mAb (M1/70) and phycoerythrin (PE)-streptavidin conjugate from BD Biosciences; rat IgG2a isotype control mAb (54447), normal goat IgG, polyclonal goat anti-mouse CD36, anti-mouse LOX-1 (lectin-type oxidised LDL receptor-1), anti-human SREC-I (scavenger receptor expressed by endothelial cells-I) Ab and PE-conjugated rat anti-mouse LOX-1 mAb (214012) from R&D Systems; polyclonal goat anti-mouse SREC-I, anti-mouse RAGE (receptor for advanced glycation end products), anti-mouse stabilin-1 and rabbit anti-mouse-stabilin-1 Ab from Santa Cruz Biotechnology; PE-conjugated donkey anti-goat IgG Ab from SouthernBiotech; rat anti-mouse CD206/mannose receptor mAb (MR5D3) and PE-conjugated goat anti-rat IgG Ab from BioLegend; horseradish peroxidase (HRP)-conjugated rabbit anti-mouse IgA, F(ab’)2 fragments of goat anti-rat IgG and donkey anti-goat IgG Ab from Rockland.

Techniques: Transfection, Binding Assay, Enzyme-linked Immunosorbent Assay

Journal: Clinical and Experimental Immunology

Article Title: Investigating the role of proinflammatory CD16 + monocytes in the pathogenesis of inflammatory bowel disease

doi: 10.1111/j.1365-2249.2010.04177.x

Figure Lengend Snippet: The peripheral blood CD14+ CD16+ monocyte population is increased in active Crohn's disease. (a) Peripheral blood mononuclear cells (PBMC) were stained for CD14, CD16 and CD36 and analysed by flow cytometry. Cells were pre-gated for high forward-scatter (FSC), side-scatter (SSC) and CD36 expression, and monocytes separated by CD14/CD16 expression. The lower panels show representative dot plots from a healthy volunteer, and Crohn's disease patients with quiescent [Crohn's disease activity index (CDAI)<150] and active (CDAI>150) inflammation. (b) The ratio of CD14+ CD16+ monocytes was increased significantly in Crohn's disease patients compared to healthy controls. *P = 0·01 (two-tailed Student's t-test with Welch's correction). (c) Further categorization of patients revealed that the CD14+ CD16+ population was increased in active Crohn's disease compared to healthy controls, but not quiescent disease or patients with ulcerative colitis. *P < 0·05 (Bonferroni's post-test following one-way analysis of variance).

Article Snippet: Slides were then incubated at 4°C overnight with the following primary antibodies: mouse anti-CD14 (BioLegend, San Diego, CA, USA), rat anti-CD16 (Santa Cruz Biotechnology, Santa Cruz, CA, USA) and rabbit anti-CD36 (Novus Biologicals, Littleton, CO, USA).

Techniques: Staining, Flow Cytometry, Expressing, Activity Assay, Two Tailed Test

Journal: Clinical and Experimental Immunology

Article Title: Investigating the role of proinflammatory CD16 + monocytes in the pathogenesis of inflammatory bowel disease

doi: 10.1111/j.1365-2249.2010.04177.x

Figure Lengend Snippet: The number of CD14+ CD16+ monocytes is enhanced significantly in the lamina propria of Crohn's disease patients. (a) Monocytic lineage of CD14+ CD16+ cells in the colonic mucosa was confirmed by three-channel immunofluorescence microscopy for CD14 (green), CD16 (red) and CD36 (blue). No or limited CD36 staining was observed in CD14+ CD16– cells (i) and CD14– CD16+ cells (ii). In contrast, CD14+ CD16+ cells stained strongly for CD36 (iii, white pseudocolour). Colonic crypts are indicated with dashed lines. Lower panels, magnification of the boxed cells shown above. (b) Representative images of colonic mucosal sections from non-inflammatory bowel disease (IBD) control patients, non-inflamed and inflamed tissue from Crohn's disease patients, and non-inflamed ulcerative colitis samples. CD14+ CD16+ monocytes are shown in yellow pseudocolour, with nuclei in blue. (c) The number of CD14+ CD16+ cells per high-power field (HPF) was significantly increased in Crohn's disease mucosa, and enhanced further in actively inflamed tissue. Each data point represents the average of five or more HPF per patient sample. *P < 0·001 versus control, †P < 0·001 versus IBD – not inflamed [Bonferroni's post-test following one-way analysis of variance (anova)]. (d) The ratio of CD14+ CD16+ cells was increased significantly in Crohn's disease patients compared to controls. *P < 0·001 (Bonferroni's post-test following one-way anova). (e) The increase in total CD14+ CD16+ monocytes was confirmed by flow cytometry. Mononuclear cells were isolated from control and Crohn's disease colonic mucosa as described in the Materials and methods, pre-gated for forward-scatter (high), side-scatter (high) CD36+, and separated by CD14/CD16 expression. A dramatic increase of CD14+ CD16+ monocytes was seen in IBD tissue (red box). Plots are representative of three patient samples per group. (f) The tumour necrosis factor (TNF)-α mean fluorescence intensity (MFI, arbitrary units) of CD14+ CD16+ monocytes was significantly higher compared to CD14+ CD16– cells in the same field of vision. The graph shows the result from 14 HPF taken from three Crohn's disease patient samples. *P < 0·001 (two-tailed Student's t-test).

Article Snippet: Slides were then incubated at 4°C overnight with the following primary antibodies: mouse anti-CD14 (BioLegend, San Diego, CA, USA), rat anti-CD16 (Santa Cruz Biotechnology, Santa Cruz, CA, USA) and rabbit anti-CD36 (Novus Biologicals, Littleton, CO, USA).

Techniques: Immunofluorescence, Microscopy, Staining, Flow Cytometry, Isolation, Expressing, Fluorescence, Two Tailed Test