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. 2021 Jul 27;14(8):734. doi: 10.3390/ph14080734

Figure 4.

Figure 4

Antibodies modulate the suppressive activity of PCL- and OVA-Mac-EVs. (A) CHS effector T cells were incubated for 30 min at 37 °C with PCL-Mac-EVs, where indicated pre-incubated with anti-TNP IgG antibodies. Afterwards, CHS effector T cells were adoptively transferred to naive recipients (n = 5 per group) that were immediately challenged with hapten to elicit CHS reaction, measured as ear swelling 24 h later. (B) CHS effector T cells were incubated for 30 min at 37 °C with PCL-Mac-EVs, where indicated pre-incubated with anti-CD9 IgG antibodies. Afterwards, CHS effector T cells were adoptively transferred to naive recipients (n = 5 per group) that were immediately challenged with hapten to elicit CHS reaction, measured as ear swelling 24 h later. (C) OT-II mouse OVA-Mac-EVs were coated onto latex beads, stained with fluoresceinated antibodies against I-A/I-E molecules, and analyzed with flow cytometry. (D) DTH effector T cells from OVA-immunized OT-II mice were incubated for 30 min at 37 °C with OT-II mouse OVA-Mac-EVs, where indicated pre-incubated with anti-OVA-323 IgG antibodies, and then were adoptively transferred to naive wild type recipients (n = 5 per group) that 24 h later were challenged with OVA to elicit DTH reaction, measured as ear swelling 24 h later. (E) After measuring of 24-h DTH ear swelling, actively immunized C57BL/6 mice (n = 5 per group) were administered intraperitoneally with OT-II mouse OVA-Mac-EVs alone or preincubated with anti-OVA-323 IgG antibodies, and subsequent DTH ear swelling was measured up to 120 h after challenge. Data are expressed as delta ± SEM. One-way or two-way ANOVA with post hoc RIR Tukey test; * p < 0.05, ** p < 0.01, *** p < 0.005, **** p < 0.001, # p < 0.05, ## p < 0.01, ### p < 0.005, #### p < 0.001.