serum ccl2  (Boster Bio)

Journal: bioRxiv

Article Title: Chemokine positioning determines mutually exclusive roles for their receptors in extravasation of pathogenic human T cells

doi: 10.1101/2023.01.25.525561

Figure Lengend Snippet: (A) Culture supernatants collected from the upper and lower wells of transwells with HUVEC monolayers were assayed for CCL2, CCL7 and CCL8 by ELISA. Data are from three experiments and bars indicate means +/− SEM. (B) Confocal microscopy images at ×40 magnification of non-permeabilized and permeabilized TNF-α + IFN-γ-stimulated HUVECs immunostained for CCL20, CCL5, CXCL9, CCL2, CCL7 or CCL8 (green) along with nuclear staining (blue) as described in Material and methods. The scale bars indicate 10 µm. Images are representative of three experiments. (C) CCR6 + CCR2 + cells were either left untreated or treated with CCL2 (100 ng/ml) just before and for 4 min after adding to the flow chambers, and CCL2-treated cells were used with or without treatment with the CCR2 antagonist, BMS22. Data points in the left panel show numbers of cells rolling, arrested and transmigrated, while the middle panel shows arrested cells as a percentage of cells rolling and the right panel shows transmigrated cells as a percentage of cells arresting. Each symbol shows data from cells from one donor, with four experiments using CCL2 and two experiments including the additional treatment with BMS22. Bars indicate means +/− SEM. p values were calculated using paired Student’s t tests, *, p<0.05; **, p<0.01; ***, p<0.001.

Article Snippet: Chemically synthesized human CCL2 and CCL5 were supplied with site-specific biotinylation (Almac).

Techniques: Enzyme-linked Immunosorbent Assay, Confocal Microscopy, Staining

Journal: bioRxiv

Article Title: Chemokine positioning determines mutually exclusive roles for their receptors in extravasation of pathogenic human T cells

doi: 10.1101/2023.01.25.525561

Figure Lengend Snippet: (A) CCL2-CXCL9 chimera sequence with signal peptide sequence shown in green, CCL2-sequence in black and C-terminal GAG-binding sequence of CXCL9 in red (top). Confocal microscopy images of control or CCL2-CXCL9 chimera transduced, TNF-α-stimulated HUVECs immunostained for CCL2 (red) and DAPI (blue) as described in Materials and methods (bottom). (B) Numbers of CCR6 + CCR2 + cells, either untreated or treated with the CCR2 antagonist, BMS22, rolling, arrested and transmigrated on TNF- α -activated HUVECs transduced with either control virus or with virus encoding the CCL2-CXCL9 chimera. Middle panel shows arrested cells as a percentage of cells rolling and in the right panel transmigrated cells as a percentage of cells arresting. Each symbol shows data for cells from one donor, with cells from four donors in each treatment group except for the BMS22-treated cells with control virus-transduced HUVECs, where cells from three donors were used. Bars indicate means +/− SEM. p values were calculated using paired t tests, *, p<0.05; **, p<0.01; ***, p<0.001.

Article Snippet: Chemically synthesized human CCL2 and CCL5 were supplied with site-specific biotinylation (Almac).

Techniques: Sequencing, Binding Assay, Confocal Microscopy, Control, Transduction, Virus

Journal: bioRxiv

Article Title: Chemokine positioning determines mutually exclusive roles for their receptors in extravasation of pathogenic human T cells

doi: 10.1101/2023.01.25.525561

Figure Lengend Snippet: Inflamed endothelial cells upregulate expression of selectins and integrin ligands and secrete various chemokines. Selectin-selectin ligand interactions help capture cells and allow them to roll along the endothelium. For pathogenic Th17 cells, endothelial cell-bound chemokines can then stimulate CCR6, CCR5 and CXCR3 to activate integrins for binding to intercellular adhesion molecules and mediate firm arrest. Secreted chemokines that fail to adhere to endothelial cells, such as CCL2 and other CCR2 ligands, are diluted by blood flow to create a transendothelial gradient that induces TEM.

Article Snippet: Chemically synthesized human CCL2 and CCL5 were supplied with site-specific biotinylation (Almac).

Techniques: Expressing, Binding Assay

Journal: Frontiers in Immunology

Article Title: Heterologous prime-boost cellular vaccination induces potent antitumor immunity against triple negative breast cancer

doi: 10.3389/fimmu.2023.1098344

Figure Lengend Snippet: ICD inducers improve the release of immunogenic mediators from TNBC cells. Cytokine and chemokine levels from (A) mouse 4T1 and human (B) BT-549 cell line culture supernatants were quantified by ELISA following treatment of with IC50 concentrations of ICD inducers or infected with 10 MOI of VSVd51 for 24h. The results were compared to non-treated cells. ELISA was performed using supernatant pooled from 3 independent experiments, where *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; (n.s., no significance).

Article Snippet: ELISA kits for detecting TNFα, IL-10, IL-1β, CCL2, CCL4, CCl5 (all Peprotech), TGFβ Elisa (Invitrogen), PGE2 (Cayman), IFN1β (R&D Systems) were performed according to manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Infection