Possible Implication of Fcγ Receptor-Mediated Trogocytosis in Susceptibility to Systemic Autoimmune Disease

Figure 2

Dynamism of molecules transferred to neutrophils via FcγR-mediated trogocytosis. Heparinized peripheral blood samples (100 μL) were made to react with 0.1 μg/mL of PE-labeled anti-CD8 Ab for 30 min at room temperature. PE-labeled mouse IgG1 was used as an isotype-matched control. After depletion of erythrocytes, the cells were incubated in RPMI-1640 medium with 10% FBS for 0–8 hours at 37°C. Subsequently, the cells were resuspended in 100 μL of PBS and then made to react with 0.1 μg of PECy5-labeled anti-CD8 Ab for 20 min at room temperature. After removal of unbounded Ab, the cells were resuspended in 100 μL of PBS followed by reaction with 0.1 μg of FITC-labeled anti-CD15 Ab for 20 min at room temperature and then served for FCM. The PECy5-labeled anti-CD8 Ab used in this experiment could recognize a different epitope from that recognized by the PE-labeled anti-CD8 Ab. Experiments were repeated at least 3 times. Similar results were reproduced.