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. 2023 Nov 29;15(1):2281763. doi: 10.1080/19420862.2023.2281763

Figure 2.

Figure 2.

CIT-013 does not bind blood cells and does not affect neutrophil anti-microbial activity other than NETosis.

(a) Binding of HiLyteTM Fluor 488-conjugated isotype control Ab (HL488-cIgG) or HL488-CIT-013 to erythrocytes, thrombocytes, T cells, B cells, monocytes, and neutrophils was measured using flow cytometry. Neutrophils stimulated with A23187 for 120 min (netting neutrophils) were used as positive control. (b) Neutrophil phagocytosis of fluorescent pHrodoTM Green S. aureus BioparticlesTM Conjugates in the absence (No Ab) or presence of cIgG, CIT-013, or Cyto D (actin polymerization inhibitor as a positive control inhibitor of phagocytosis). (c) ROS production by neutrophils stimulated with Ig-opsonized heat-killed S. aureus in the absence or presence of cIgG, CIT-013, or DPI (NADPH oxidase inhibitor as a positive control inhibitor for ROS production) (n = 6). Statistics were performed on the area under the curve. (d) CD11b membrane degranulation marker on neutrophils stimulated with LPS in the absence or presence of cIgG, CIT-013, or Cyto D (positive control inhibitor of degranulation) at t = 180 min. *P<.05, ***P<.001, and ****P<.0001, two-tailed paired t test (a), two-tailed unpaired t test (b), and RM one-way ANOVA using Dunnett’s multiple comparisons test (c). RFU = relative fluorescence units; MFI = mean fluorescence intensity.