antibodies against cxcl10  (R&D Systems)

Journal: bioRxiv

Article Title: Microengineered transplantation of human solid tumors for in vitro studies of CAR T immunotherapy

doi: 10.1101/2025.04.25.650709

Figure Lengend Snippet: a, Conceptual diagram illustrating pairwise mapping of directional ligand-receptor interactions between different cell types in the engineered tumor model. b,c, Chord diagrams of cell pair-specific ligand-receptor interactions with adjusted p-values < 0.05 and total mean expression > 0.35. d, Visualization of interactions between CAR T cells (CAR Ts) and endothelial cells (Endos) mediated by CXCR3/DPP4-CXCL10/CXCL11. e, Violin plots comparing the expression of these mediators across all cell types. f, Experimental timeline for CAR T cell infusion and drug treatment. g, Representative confocal micrographs of single tumors treated with CAR T cells and different concentrations of LAF237. Blood vessels are not shown in these images. Scale bars, 250 μm. h, Representative images of CAR T cell-infused tumors at Day 26. Scale bars, 200 µm. i, Quantification and comparison of tumor area (top) and CAR T cell-occupied hydrogel area (bottom). Shaded in grey is a period of CAR T cell infusion and daily LAF237 treatment. Data are presented as mean ± SEM (n ≥ 3). j,k, Quantification of CXCL10 and CXCL11 levels ( j ) and the activity of DPP4 ( k ) in device effluent. Shaded in grey is a period of CAR T cell infusion and daily LAF237 treatment. Data are presented as mean ± SEM in ( j ) and as mean ± SD in ( k ) (n ≥ 3). CAR T cells derived from three healthy donors were tested in this study.

Article Snippet: The membranes were blocked with 5% skim milk for 1 hour at room temperature, followed by overnight incubation at 4°C with primary antibodies against CXCL10 (MAB266-100, R&D systems).

Techniques: Expressing, Comparison, Activity Assay, Derivative Assay

Journal: Cells

Article Title: Induction of CXCL10-Mediated Cell Migration by Different Types of Galectins

doi: 10.3390/cells10020274

Figure Lengend Snippet: Galectins induce CXCL10 in a cell type-specific manner. ( A ) SDS-polyacrylamide gel electrophoresis of recombinant human galectins (rhGal) stained with Coomassie Blue. Molecular weights of protein standards are indicated on the left. The hemagglutination assay was performed using serial two-fold dilutions of the recombinant protein. The bottom row contains protein incubated with β-lactose, a competitive carbohydrate inhibitor of galectin binding. ( B ) Cultures of human corneal fibroblasts, human corneal epithelial cells, THP-1 monocytes and human umbilical vein endothelial cells (HUVECs) were incubated with 50 µg/mL rhGal-1, -3 and -8 or 50 µg/mL IFN-γ. Quantitative real-time PCR was used to determine the levels of CXCL10 mRNA after 6 h of incubation ( n = 3 independent experiments), whereas ELISA was used to determine the levels of CXCL10 protein in cell culture supernatants after 24 h of incubation ( n = 6 independent experiments). The data represent the mean ± SEM.

Article Snippet: A mouse monoclonal antibody against CXCL10 (1.2 μg/mL; MAB266, R&D Systems) was added to the lower compartment of the chamber in neutralization studies.

Techniques: Polyacrylamide Gel Electrophoresis, Recombinant, Staining, Hemagglutination Assay, Incubation, Binding Assay, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: Cells

Article Title: Induction of CXCL10-Mediated Cell Migration by Different Types of Galectins

doi: 10.3390/cells10020274

Figure Lengend Snippet: Galectins differentially dictate fibroblast-dependent immune cell chemotaxis. ( A ) Recombinant human galectin (rhGal)-1, -3 and -8 were incubated with fibroblasts for 24 h. The conditioned media were pre-cleared with β-lactose Sepharose beads three times to remove galectins prior to chemotaxis assays. The presence of galectins in pre-cleared conditioned media was evaluated by immunoblotting. ( B ) Quantification by flow cytometry of the migration of CFSE-labeled THP-1 and Jurkat cells to conditioned media from fibroblasts treated for 24 h with increasing concentrations of recombinant galectins or BSA ( n = 3 independent experiments performed in duplicate). ( C ) Migration of CFSE-labeled THP-1 and Jurkat cells to conditioned media from fibroblasts treated for 24 h with 50 μg/mL of recombinant galectins or BSA. A CXCL10 neutralizing antibody was added to the conditioned media in neutralization studies ( n = 4–5 independent experiments in duplicate). The data in ( B ) represent the mean ± SEM. The box and whisker plots show the 25 and 75 percentiles (box), the median, and the minimum and maximum data values (whiskers). Significance was determined using Student’s t test. ** p < 0.01.

Article Snippet: A mouse monoclonal antibody against CXCL10 (1.2 μg/mL; MAB266, R&D Systems) was added to the lower compartment of the chamber in neutralization studies.

Techniques: Chemotaxis Assay, Recombinant, Incubation, Western Blot, Flow Cytometry, Migration, Labeling, Neutralization, Whisker Assay

Journal: PLoS ONE

Article Title: Human Primary Adipocytes Exhibit Immune Cell Function: Adipocytes Prime Inflammation Independent of Macrophages

doi: 10.1371/journal.pone.0017154

Figure Lengend Snippet: Panel A shows the analysis of CD4+ cells migration caused by adipocytes-produced chemokines after 10 h incubation. The lower compartment (insert) contained attached adipocytes treated with LPS (200 ng/ml) or untreated, as well as antibodies against MCP-1 and IP-10 (2 µg/ml) to neutralize the effect of these two chemo-attractants. The upper chamber (insert) contained purified and rested CD4+ T cells. The cells were incubated at 37°C for 10 h. The inserts were then removed and the cells migrated toward the adipocytes were collected in culture medium without serum. The number of migrated cells was obtained by cell counting using a Bürker-Türk chamber. This figure represents two independent experiments with similar results (only differ in the number of migrated cells). Each experiment was done in triplicate. Error bars show standard deviations of means for triplicate cultures per condition. The p-value obtained from an independent two-tailed test samples (T-test). P<0.05 was accepted as statistically significant. The controls in the migration assay were 1- culture medium without serum, 2- culture medium serum free with LPS, and culture medium serum free with IgG isotype (this is not shown in Figure 10 A and 10 B , because this was executed separately and the obtained results were the same as culture medium serum free with LPS). Panel B shows the analysis of CD4+ cells migration caused by adipocyte-produced chemokines after 30 h incubation. All cell migration conditions were similar to the conditions described in panel A , except that the cells were incubated at 37°C for 30 h.

Article Snippet: One migration experiment was performed with the plated CD4+ cells (7×10 5 ) suspension on top of a culture insert in the absence of serum and incubated at 37°C for 30 h. Monoclonal antibodies against human MCP-1 and IP-10 (R & D system) were used to neutralize the effect of these two chemoattractants in migration assay.

Techniques: Migration, Produced, Incubation, Purification, Cell Counting, Two Tailed Test