Journal: Oncotarget

Article Title: Clinical relevance of circulating mucosal-associated invariant T cell levels and their anti-cancer activity in patients with mucosal-associated cancer

doi: 10.18632/oncotarget.11187

Figure Lengend Snippet: Purified T cells (5 × 10 5 cells/well) were added to upper chambers of Transwell plates. rhCCL20 (200 ng/ml), rhCXCL16 (200 ng/ml), mixture of rhCCL20 and rhCXCL16 (200 ng/ml), rhCCL21 (200 ng/ml), and PBS as a negative control were added to lower chambers. Plates were incubated for 90 minutes at 37°C. Migrated cells were stained with APC-Alexa Fluor 750-conjugated anti-CD3, FITC-conjugated anti-TCR γδ, APC-conjugated anti-TCR Vα7.2 and PE-Cy5-conjugated anti-CD161 mAbs and then counted using Neubauer chamber and flow cytometer. Migration index was calculated by the number of cells that migrated in the presence of each chemokine divided by the number of cells that migrated in the presence of PBS. The migration index of MAIT cells was normalized to the migration index of T cells. Values are expressed as the mean ± SEM. Data were obtained from 5 HCs. *, P < 0.05, **, P < 0.005 by paired t test.

Article Snippet: Recombinant human (rh) CCL20 (200 ng/ml; PeproTech, London, UA), rhCXCL16 (200 ng/ml; PeproTech), mixture of rhCCL20 and rhCXCL16 (200 ng/ml), rhCCL21 (200 ng/ml; PeproTech) and PBS were added to lower chambers containing serum free RPMI1640 media.

Techniques: Purification, Negative Control, Incubation, Staining, Flow Cytometry, Migration

Journal: PLoS ONE

Article Title: A humanized monoclonal antibody against the endothelial chemokine CCL21 for the diagnosis and treatment of inflammatory bowel disease

doi: 10.1371/journal.pone.0252805

Figure Lengend Snippet: (A) Inhibition of migration in transwell chemotaxis assays of helper T cells (CD3 + /CD4 + ) towards 1200 ng/ml rhCCL21 by the 16 humanized clones, referred to as V1 to V16, used at 100 μg/ml. All versions of humanized clones tested inhibited T cell migration towards CCL21 to some degree. However, V6 was the most potent at inhibiting T cell migration towards CCL21 (arrow). Data are the mean ± SD of triplicate wells. One-way ANOVA with post-hoc Tukey’s multiple comparison tests were performed for statistical analysis. All groups had a p<0.0001 when compared to control. (B-D) Confirmation of V6 inhibition of CD3 + /CD4 + /CD8 - /CCR7 + /CD45RA + /CD27 + Naïve Th-cell chemotaxis towards 100, 500, 800, 1200, and 1600 ng/ml rhCCL21 using CD3 + peripheral blood naïve Th-cells from 3 normal human donors. Data are the mean ± SD of triplicate wells.

Article Snippet: Costar 24-well transwell plates with 6.5 mm diameter inserts with 5.0 μm pores (Sigma-Aldrich, St. Louis, MO) were prepared by placing 650 μl of prewarmed RPMI with 0.5% BSA that contained 0, 1200 ng/mL rhCCL21 (R&D Systems, Minneapolis, MN) or 1200ng/mL rhCCL21 pretreated for 1 hour with sample clones #7–39 in the bottom well and allowing plates to acclimate at 37°C for half an hour prior to chemotaxis assay.

Techniques: Inhibition, Migration, Chemotaxis Assay, Clone Assay, Comparison