Figure 2.
CD45-ADC conditioning at lower dose (1.5 mg/kg, d2) facilitated complete donor chimerism in a minor–mismatch transplant model. (A-C) Female or male Fanca−/− mice (10- to 12-week-old) were conditioned with isotype-ADC (0.5 mg/kg) or CD45-ADC (0.5 or 1.5 mg/kg) on d2. Depletion of HSPCs in BM (A), and hematopoietic cells and adaptive immune cells in spleen (B) and thymus (C) were assessed by flow cytometry on day 0. Untreated mice served as control. (A) Pooled data from 2 experiments are shown (n = 7-8 mice per group). For panels B-C, experiments were performed twice, and data from 1 representative experiment are shown (n = 3-4 mice per group). For panels A-C, data represent mean ± SEM. ∗P < .05, ∗∗P < .01, and ∗∗∗P < .001 using the 2-tailed Student t test. (D-I) Female or male Fanca−/− mice (10- to 16-week-old) were conditioned with isotype-ADC (0.5 mg/kg, n = 7; or 1.5 mg/kg, n = 12) or CD45-ADC (1.5 mg/kg, n = 20) on d2 and received 40 × 106 donor (H-2b, CD45.1) BM cells on d0. (D) Engraftment of donor cells (CD45.1) in the PB of mice who underwent transplantation were analyzed at 5, 10, and 25 weeks post-BMT by flow cytometry, and multilineage PB donor chimerism was analyzed at 25 weeks post-BMT (E). For panels D-E, data represent mean ± SEM. ∗P < .05 and ∗∗∗∗P < .0001 using the 2-tailed Student t test. Pooled data from 2 experiments are shown. (F-I) Recipients were killed (n = 4-6 mice per group) at 30 weeks post-BMT and donor (CD45.1+) chimerism in BM (F,I), thymus (G), and spleen (H) were analyzed by flow cytometry. Experiments were performed twice, and data from 1 representative experiment are shown. Data represent mean ± SEM. ∗∗P < .01, ∗∗∗P < .001, and ∗∗∗∗P < .0001 using the 2-tailed Student t test. (J-K) WT B6 (H-2b, CD45.2) mice received lethal TBI (900 cGy) on d1. CD45.1+ (B6) donor BM cells were isolated at 26 weeks post-BMT from primary Fanca−/− recipients conditioned with CD45-ADC (1.5 or 3 mg/kg). For adoptive transfer into secondary recipients, BM cells (10 × 106) from primary recipients were infused into lethally irradiated WT B6 recipients (n = 7-8 mice per group) on d0. CD45.1+ (B6) donor BM cells isolated from naïve mice and infused (10 × 106) into lethally irradiated WT B6 recipients (n = 8 mice) served as control. (J) Engraftment of donor cells (CD45.1) in PB of secondary WT recipients were analyzed at 5, 10, and 18 weeks post-BMT and multilineage PB donor chimerism was analyzed at 18 weeks post-BMT (K). For panels J-K, data represent mean ± SEM. ∗P < .05, ∗∗P < .01, and ∗∗∗P < .001 using the 2-tailed Student t test.

CD45-ADC conditioning at lower dose (1.5 mg/kg, d2) facilitated complete donor chimerism in a minor–mismatch transplant model. (A-C) Female or male Fanca−/− mice (10- to 12-week-old) were conditioned with isotype-ADC (0.5 mg/kg) or CD45-ADC (0.5 or 1.5 mg/kg) on d2. Depletion of HSPCs in BM (A), and hematopoietic cells and adaptive immune cells in spleen (B) and thymus (C) were assessed by flow cytometry on day 0. Untreated mice served as control. (A) Pooled data from 2 experiments are shown (n = 7-8 mice per group). For panels B-C, experiments were performed twice, and data from 1 representative experiment are shown (n = 3-4 mice per group). For panels A-C, data represent mean ± SEM. ∗P < .05, ∗∗P < .01, and ∗∗∗P < .001 using the 2-tailed Student t test. (D-I) Female or male Fanca−/− mice (10- to 16-week-old) were conditioned with isotype-ADC (0.5 mg/kg, n = 7; or 1.5 mg/kg, n = 12) or CD45-ADC (1.5 mg/kg, n = 20) on d2 and received 40 × 106 donor (H-2b, CD45.1) BM cells on d0. (D) Engraftment of donor cells (CD45.1) in the PB of mice who underwent transplantation were analyzed at 5, 10, and 25 weeks post-BMT by flow cytometry, and multilineage PB donor chimerism was analyzed at 25 weeks post-BMT (E). For panels D-E, data represent mean ± SEM. ∗P < .05 and ∗∗∗∗P < .0001 using the 2-tailed Student t test. Pooled data from 2 experiments are shown. (F-I) Recipients were killed (n = 4-6 mice per group) at 30 weeks post-BMT and donor (CD45.1+) chimerism in BM (F,I), thymus (G), and spleen (H) were analyzed by flow cytometry. Experiments were performed twice, and data from 1 representative experiment are shown. Data represent mean ± SEM. ∗∗P < .01, ∗∗∗P < .001, and ∗∗∗∗P < .0001 using the 2-tailed Student t test. (J-K) WT B6 (H-2b, CD45.2) mice received lethal TBI (900 cGy) on d1. CD45.1+ (B6) donor BM cells were isolated at 26 weeks post-BMT from primary Fanca−/− recipients conditioned with CD45-ADC (1.5 or 3 mg/kg). For adoptive transfer into secondary recipients, BM cells (10 × 106) from primary recipients were infused into lethally irradiated WT B6 recipients (n = 7-8 mice per group) on d0. CD45.1+ (B6) donor BM cells isolated from naïve mice and infused (10 × 106) into lethally irradiated WT B6 recipients (n = 8 mice) served as control. (J) Engraftment of donor cells (CD45.1) in PB of secondary WT recipients were analyzed at 5, 10, and 18 weeks post-BMT and multilineage PB donor chimerism was analyzed at 18 weeks post-BMT (K). For panels J-K, data represent mean ± SEM. ∗P < .05, ∗∗P < .01, and ∗∗∗P < .001 using the 2-tailed Student t test.

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