American Society of Hematology

Figure 3

$Figure 3. Removal of donor CD4+ T cells decreases the risk of delayed bortezomib-related lethal toxicity. (A) B6 (H2b) recipients of 15 million BALB/c (H2d) bone marrow and 12 million SCs from donors depleted of CD4+ or CD8+ T cells in vivo, before tissue collection, were treated with 15 μg bortezomib per dose on day +5 and day +16 (n = 6 mice/group) and monitored for survival. Representative data from 1 of 2 independent experiments are shown. (B) BALB/c (H2d) recipients of 10 million FvB (H2q) bone marrow and 2 million SCs from donors depleted of CD4+ or CD8+ T cells in vivo, before tissue collection, were treated with 7.5 μg bortezomib on day +6 (n = 6 mice/group) and monitored for survival. (C) BALB/c (H2d) recipients of 10 million FvB (H2q) bone marrow and 2 million SCs were treated with indicated depleting mAb on day +4 and with 7.5 μg bortezomib per dose on day +6 after BMT (n = 5 mice/group) and monitored for survival. (D,F) B6 recipients of BALB/c cells underwent transplantation as described for panel A. Serum was collected on day +6 after BMT (n = 3-7 mice/group) and analyzed for TNFα and IFNγ. Representative data from 1 of 2 independent experiments are shown as means plus or minus SEM. (E,G) BALB/c recipients of 10 million FvB BMCs and 3 million SCs from donors that received CD4 and/or CD8 depleting antibody before tissue collection and treated as described in panel B. Serum was collected on day +6 after BMT (n = 5 mice/group) and analyzed for TNFα and IFNγ. Results are shown as means plus or minus SEM; representative data from 1 of 2 independent experiments are shown. (H) BALB/c recipients of FvB T cell–depleted bone marrow and 3 million SCs from untreated donors or in vivo CD4+ cell–depleted donors were treated with 10 μg bortezomib or PBS on day +6 and the ileum was collected for RT-qPCR analysis on day +7 (n = 3-6 mice/group). Data are expressed by the ΔΔCt method in which TNFR1 gene expression is normalized to ACTB in individual samples and then relative expression is compared with the mean value from the group receiving unfractionated spleen cells and PBS. Significant increases in TNFR1 gene expression are shown as means (± SEM; P < .05; Student t test).$

Removal of donor CD4⁺ T cells decreases the risk of delayed bortezomib-related lethal toxicity. (A) B6 (H2^b) recipients of 15 million BALB/c (H2^d) bone marrow and 12 million SCs from donors depleted of CD4⁺ or CD8⁺ T cells in vivo, before tissue collection, were treated with 15 μg bortezomib per dose on day +5 and day +16 (n = 6 mice/group) and monitored for survival. Representative data from 1 of 2 independent experiments are shown. (B) BALB/c (H2^d) recipients of 10 million FvB (H2^q) bone marrow and 2 million SCs from donors depleted of CD4⁺ or CD8⁺ T cells in vivo, before tissue collection, were treated with 7.5 μg bortezomib on day +6 (n = 6 mice/group) and monitored for survival. (C) BALB/c (H2^d) recipients of 10 million FvB (H2^q) bone marrow and 2 million SCs were treated with indicated depleting mAb on day +4 and with 7.5 μg bortezomib per dose on day +6 after BMT (n = 5 mice/group) and monitored for survival. (D,F) B6 recipients of BALB/c cells underwent transplantation as described for panel A. Serum was collected on day +6 after BMT (n = 3-7 mice/group) and analyzed for TNFα and IFNγ. Representative data from 1 of 2 independent experiments are shown as means plus or minus SEM. (E,G) BALB/c recipients of 10 million FvB BMCs and 3 million SCs from donors that received CD4 and/or CD8 depleting antibody before tissue collection and treated as described in panel B. Serum was collected on day +6 after BMT (n = 5 mice/group) and analyzed for TNFα and IFNγ. Results are shown as means plus or minus SEM; representative data from 1 of 2 independent experiments are shown. (H) BALB/c recipients of FvB T cell–depleted bone marrow and 3 million SCs from untreated donors or in vivo CD4⁺ cell–depleted donors were treated with 10 μg bortezomib or PBS on day +6 and the ileum was collected for RT-qPCR analysis on day +7 (n = 3-6 mice/group). Data are expressed by the ΔΔCt method in which TNFR1 gene expression is normalized to ACTB in individual samples and then relative expression is compared with the mean value from the group receiving unfractionated spleen cells and PBS. Significant increases in TNFR1 gene expression are shown as means (± SEM; P < .05; Student t test).

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