Figure 3.
Conditioning with lower dose CD45-ADC (1.5 mg/kg) is associated with mild, transient hepatotoxicity in Fanca−/− recipients. (A) Unconjugated CD45.2 antibody conditioning could not deplete HSPCs in BM. Female WT B6 mice (10 weeks old) were conditioned with isotype-ADC (0.5 mg/kg) or CD45-ADC (1.5 mg/kg) on d2. A cohort of B6 mice were treated with unconjugated CD45.2 antibody (1 μg/gm per day, clone 104, mouse IgG2a) daily from d6 through d3. Depletion of HSPCs in BM were assessed on d0 by flow cytometry. Untreated B6 mice served as control. Data from 1 experiment are shown (n = 4 mice per group). Data represent mean ± SEM. ∗∗P < .01, ∗∗∗P < .001, and ∗∗∗∗P < .0001 using the 2-tailed Student t test. (B-F) Conditioning with lower dose of CD45-ADC (1.5 mg/kg) caused mild, transient hepatotoxicity in Fanca−/− recipients. Female or male Fanca−/− mice (10- to 12-week-old) were conditioned with isotype-ADC (0.5 mg/kg) or CD45-ADC (1.5 mg/kg) on d2 and then received 40 × 106 donor (H-2b, CD45.1) BM cells on d0. (B) Engraftment of donor cells (CD45.1) in PB of mice that underwent transplant, and PB donor and host cell subsets were analyzed by flow cytometry at 10 weeks post-BMT. (C-D) Blood samples were collected at d1, d6, and d11 post-BMT. In a second experiment, blood samples were also collected at d0 before BM infusion and at d28 and d56 post-BMT. The levels of ALT, AST, albumin, and bilirubin in serum samples were measured. Untreated mice served as control. ∗P < .05, ∗∗P < .01, and ∗∗∗P < .001 vs untreated mice. #; isotype-ADC vs CD45-ADC, P < .05. (E) Blood samples were collected at d1, d6, and d11 post-BMT and levels of tumor necrosis factor-α or IFN-γ in serum samples were measured. Untreated mice served as control. ∗P < .05 and ∗∗P < .01, isotype-ADC vs CD45–ADC-conditioned mice. #; IFN-γ (d11): untreated control vs CD45-ADC, P < .05. (F) Representative mice were euthanized at d12 or d78 post-BMT and harvested organs were processed for hematoxylin and eosin staining and histopathological assessment was performed. For panels B-F, data represent mean ± SEM (n = 5-6 mice per group per time point). ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, and ∗∗∗∗P < .0001 using the 2-tailed Student t test.

Conditioning with lower dose CD45-ADC (1.5 mg/kg) is associated with mild, transient hepatotoxicity in Fanca−/− recipients. (A) Unconjugated CD45.2 antibody conditioning could not deplete HSPCs in BM. Female WT B6 mice (10 weeks old) were conditioned with isotype-ADC (0.5 mg/kg) or CD45-ADC (1.5 mg/kg) on d2. A cohort of B6 mice were treated with unconjugated CD45.2 antibody (1 μg/gm per day, clone 104, mouse IgG2a) daily from d6 through d3. Depletion of HSPCs in BM were assessed on d0 by flow cytometry. Untreated B6 mice served as control. Data from 1 experiment are shown (n = 4 mice per group). Data represent mean ± SEM. ∗∗P < .01, ∗∗∗P < .001, and ∗∗∗∗P < .0001 using the 2-tailed Student t test. (B-F) Conditioning with lower dose of CD45-ADC (1.5 mg/kg) caused mild, transient hepatotoxicity in Fanca−/− recipients. Female or male Fanca−/− mice (10- to 12-week-old) were conditioned with isotype-ADC (0.5 mg/kg) or CD45-ADC (1.5 mg/kg) on d2 and then received 40 × 106 donor (H-2b, CD45.1) BM cells on d0. (B) Engraftment of donor cells (CD45.1) in PB of mice that underwent transplant, and PB donor and host cell subsets were analyzed by flow cytometry at 10 weeks post-BMT. (C-D) Blood samples were collected at d1, d6, and d11 post-BMT. In a second experiment, blood samples were also collected at d0 before BM infusion and at d28 and d56 post-BMT. The levels of ALT, AST, albumin, and bilirubin in serum samples were measured. Untreated mice served as control. ∗P < .05, ∗∗P < .01, and ∗∗∗P < .001 vs untreated mice. #; isotype-ADC vs CD45-ADC, P < .05. (E) Blood samples were collected at d1, d6, and d11 post-BMT and levels of tumor necrosis factor-α or IFN-γ in serum samples were measured. Untreated mice served as control. ∗P < .05 and ∗∗P < .01, isotype-ADC vs CD45–ADC-conditioned mice. #; IFN-γ (d11): untreated control vs CD45-ADC, P < .05. (F) Representative mice were euthanized at d12 or d78 post-BMT and harvested organs were processed for hematoxylin and eosin staining and histopathological assessment was performed. For panels B-F, data represent mean ± SEM (n = 5-6 mice per group per time point). ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, and ∗∗∗∗P < .0001 using the 2-tailed Student t test.

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