Influence and interaction of PMap with monocytes.
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CCL2 (50 ng/μl) was used as a positive control.
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Chemokine (C-C motif) receptor 2 (CCR2) ligand promotes proliferation of human and murine blood fibrocytes.
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Human blood fibrocytes were stimulated with human chemokine (C-C motif) ligand 2 (hCCL2) or platelet-derived growth factor -AB and murine blood fibrocytes (pooled blood from n = 10 mice) were stimulated with mCCL2, mCCL12 or…
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Human blood fibrocytes were stimulated with human chemokine (C-C motif) ligand 2 (hCCL2) or platelet-derived growth factor -AB and murine blood fibrocytes (pooled blood from n = 10 mice) were stimulated with mCCL2, mCCL12 or PDGF-AB.
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A–D: typical responses of neuritis-treated C-fiber axons to CCL2 and TNF-α application (A: CCL2 silent; B: CCL2 ongoing; C: TNF-α silent; D: TNF-α ongoing).
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E: interspike-interval plot for a C-fiber axon with baseline ongoing activity that shows the transient response to CCL2 application.
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Duffy antigen co-localizes with caveolin-1 but not with clathrin.
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The DIH cells were stimulated with or without CCL2 100 ng/ml for 120 minutes.
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Degradation of MCPs
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Recombinant human CCL2, CCL7, CCL8 and CCL13 possessing an N-terminal glutaminyl residue were dissolved in 25 mM Tris/HCl pH 7.6 at a final concentration of 10 µg/ml.
- MCPs were either pre-incubated with recombinant human QC (0.6 µg/mlet…
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Recombinant human CCL2, CCL7, CCL8 and CCL13 possessing an N-terminal glutaminyl residue were dissolved in 25 mM Tris/HCl pH 7.6 at a final concentration of 10 µg/ml.
- MCPs were either pre-incubated with recombinant human QC (0.6 µg/mlet al, 1-76) was either pre-incubated with recombinant human QC and subsequently incubated with human plasma or incubated with human plasma without QC.
- All samples were analysed using Maldi-TOF-MS.
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Cytokine and Chemokine Measurement
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TNFα, TNFβ, IFNγ, IL-10, IL-12,IL-17 (all&, ), IL-5 and MCP-1 were measured in human tonsil cell culture media using enzyme-linked immunosorbent assay (ELISA).
- Murine cytokine and chemokines were measured in infected serum or thigh homogenate using…
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TNFα, TNFβ, IFNγ, IL-10, IL-12,IL-17 (all&, ), IL-5 and MCP-1 were measured in human tonsil cell culture media using enzyme-linked immunosorbent assay (ELISA).
- Murine cytokine and chemokines were measured in infected serum or thigh homogenate using a mouse cytokine Luminex® 20-plex panel and analyzed on a Bio-Rad Bio-Plex 200 system.
- The cytokines and chemokines measured were as follows; TNFα,GMCSF, FGF, VEGF, IL-1α, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12, IL-13, IL-17, MIP-1α, MCP-1, IFNγ, IP-10, MIG, and KC.
- For analysis, samples below the lowest level of detected were assigned a value half of the lowest measurable value.
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Dendritic cells are largely perivascular indicating transmigration to CCL2 producing experimental autoimmune encephalomyelitis lesion areas.
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In all fields, CCL2 staining (green) was closely associated with myelin basic protein staining (red).
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Human lung cancer cells produce variable amounts of CCL2 and show sensitivity to cytocidal activity mediated by CD8+ T cells genetically engineered to express WT1-specific TCR.
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ELISA assay revealed that 10 human lung cancer cell lines examined produced various amounts of CCL2 in the culture supernatant.
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Drugs
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TMZ, SAHA and the JNK-inhibitor SP600125 were primarily dissolved in DMSO and diluted in water.
- For in vivo studies, SAHA was dissolved in DMSO/ethanol/PBS (10:5:85) and administered in a volume of 10 mL/kg.
- Human CCL2/MCP-1 monoclonal neutralizing antibody…
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TMZ, SAHA and the JNK-inhibitor SP600125 were primarily dissolved in DMSO and diluted in water.
- For in vivo studies, SAHA was dissolved in DMSO/ethanol/PBS (10:5:85) and administered in a volume of 10 mL/kg.
- Human CCL2/MCP-1 monoclonal neutralizing antibody [
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Cell Culture and Treatments
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In experiments investigating CXC-chemokine signaling, cells were incubated in serum-free, phenol-red free RMPI 1640 or medium'>DMEM medium for 16h prior to exposure to 3nM recombinant human (rh)-CXCL8, 100ng/ml rh-CXCL12 or 100ng/ml rh-CCL2 .
- For inhibition…
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In experiments investigating CXC-chemokine signaling, cells were incubated in serum-free, phenol-red free RMPI 1640 or medium'>DMEM medium for 16h prior to exposure to 3nM recombinant human (rh)-CXCL8, 100ng/ml rh-CXCL12 or 100ng/ml rh-CCL2 .
- For inhibition of chemokine signaling, cells were treated with RS102895 (CCR2 antagonist, 10nM, , ), AMD3100 (CXCR4 antagonist, 25μg/ml) or a CXCR1/2-derived pepducin (x1/2pal-i3, 300nM, , ).
- Control cells were treated with the appropriate vehicle or NT-pepducin (x1/2pal-con) sequence as required.
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