Skip to main content
. 2024 Jul 4;12(7):e008677. doi: 10.1136/jitc-2023-008677

Figure 3.

Figure 3

Anti-hOX40 mIgG1 mAb requires both activating and inhibitory FcγR for activity. hOX40het OT-I cells were transferred into transferred into hOX40KIhom, hOX40KIhomγ chain KO or hOX40KIhom FcγRIIB KO mice and then immunized as in figure 2A. (A) Response to anti-hOX40 mIgG1 (left panel) or anti-hOX40 mIgG2a (right panel) n=4 except isotype mIgG2a in hOX40KIhom where n=5. One mouse was excluded from anti-hOX40 mIgG1 in hOX40KIhom recipients due to a lack of OT-I response hence n=4. Representative of two independent experiments. (B) Peak OT-I responses in blood from hOX40KIhom and hOX40KIhomγ chain KO recipients n=5 except Isotype mIgG1 in both hOX40KIhom and hOX40KIhomγ chain KO where n=4. One of two independent experiments. Splenic analysis of OT-I (C) and CD4+Foxp3+ (D) T cells on day 4 post OVA (5 mg) and antibody (100 µg) challenge. N=4 except for anti-hOX40 mIgG2a in hOX40KI where n=3. One mouse excluded in isotype mIgG2a group in hOX40KIhom due to lack of staining hence n=3. Representative of two independent experiments. (E) OT-I kinetic responses in hOX40KIhom and hOX40KIhom FcγRIIBKO mice in response to anti-hOX40 mIgG1 (left panel) or anti-hOX40 mIgG2a (right panel) n=4 except Isotype mIgG1 in hOX40KIhom and Isotype mIgG2a in hOX40KIhom FcγRIIB KO mice where n=3. Representative of two independent experiments. (F–H) Splenic analysis of OT-I (F) and CD4+Foxp3+ (G) T cells on day 4 post ovalbumin (5 mg) and antibody (100 µg) challenge. N=3 for all hOX40KI recipient groups, n=4 for all hOX40KIhom FcγRIIB KO except for anti-hOX40 mIgG1 and anti-hOX40 mIgG2a treated groups where one mouse was excluded as due to lack of OT-I response hence n=3. One of two independent experiments. (H) Treg fold induction pooled from two independent experiments, n=7 for hOX40KIhom anti-hOX40 mIgG1 and mIgG2a, n=6 for hOX40KIhom FcγRIIB KO anti-hOX40 mIgG1 and mIgG2a. ****p<0.0001, ***p<0.001, **p<0.01, *p<0.05 mean±SEM B–D, and F and G—one-way analysis of variance with Sidak’s multiple comparison. mAb, monoclonal antibody; FcγR, Fc gamma receptors; KI, knock-in; KO, knock-out; Treg, regulatory T cells.