Mouse anti human cd8 apc

For chemotaxis assays, total peripheral blood mononuclear cells (PBMCs) were isolated from human blood and were added to a polycarbonate filter with 5 µm pores (ChemoTx, Neuro Probe Inc., Gaithersburg, MD, USA) at the concentration of 1.10⁶ cells/well. The filter was gently placed on wells that were filled with human aortic conditioned medium. Following 4 hours of migration at 37°C, cells that had migrated through the filter towards the bottom wells were harvested and stained with mouse anti-human CD3-eFluor 700, mouse anti-human CD4-PE-Cy7, mouse anti-human CD8-APC, mouse anti-human CD45RO-PE-CF594, mouse anti-human CD45RA-V450, and mouse anti-human CD19-AlexiaFluor700, all from BD Biosciences. Cells were then analyzed by flow cytometry, as described above.

All samples were acquired on a CytoFLEX S (Beckman Coulter, Indianapolis, IN), and data was analyzed using Kaluza 2.1 Flow Analysis. Software (Beckman Coulter Life Sciences). Staining for cell surface markers was carried out by incubating with antibodies for 30 min on ice. Antibodies involved in this study included Recombinant PE Anti-Mesothelin antibody(Abcam, Grand Island, NY), mouse IgG1 Isotype Control(R&D, Minneapolis, MN, USA), mouse F(ab)2 IgG (H+L) APC-conjugated Antibody (R&D, Minneapolis, MN, USA), human CD155/PVR PE-conjugated Antibody (R&D, Minneapolis, MN, USA), human TIGIT APC-conjugated Antibody (R&D, Minneapolis, MN, USA), CD226 (DNAM-1) Monoclonal Antibody, APC (eBioscience, San Diego, CA), mouse anti-human CD4-PE (BD, San Diego, CA), mouse anti-human CD8-APC (BD, San Diego, CA), mouse anti-human Foxp3-APC (BD, San Diego, CA). APC anti-mouse CD279 (PD-1) Antibody (biolegend, California, USA), PE anti-mouse CD197 (CCR7) Antibody (biolegend, California, USA), APC anti-human CD45RA Antibody (biolegend, California, USA).