Figure 2
Figure 2. KRN7000 induces hyperacute GVHD in multiple models. (A) Irradiated B6D2F1 received whole spleen from G-CSF–mobilized syngeneic B6D2F1 or allogeneic B6 donors, followed by control diluent, C20:2 at 2 μg or 5 μg per mouse, or KRN7000 at 0.5 μg or 2 μg per mouse IP on days +1 and +4. Overall survival by Kaplan-Meier analysis of combined data from 2 experiments (control allo, n = 14; C20:2 2 μg allo, n = 12; C20:2 5 μg allo, n = 16; KRN7000 2 μg allo, n = 10; KRN7000 0.5 μg allo, n = 16; KRN7000 syn, n = 8). **P < .001 for KRN7000 2 μg allo vs C20:2 2 μg and 5 μg allo. *P < .04 for KRN7000 0.5 μg allo vs C20:2 2 μg and 5 μg allo. (B) Irradiated B6 mice received whole or TCD spleen from G-CSF–mobilized BALB/c donors, followed by control diluent, C20:2, or KRN7000 IP on days +1 and +4. Overall survival by Kaplan-Meier analysis and mean ± SE of clinical scores in surviving mice at the end of each experiment (allo groups, n = 20 each; TCD group, n = 12). Combined data from 3 identical experiments. **P < .02 for C20:2 allo vs control and KRN7000 allo. *P < .05 for control allo clinical scores vs C20:2 and KRN7000 allo. (C) Leukemic relapse in irradiated B6 mice receiving whole or TCD spleen from G-CSF–mobilized BALB/c donors with the addition of host-type luciferase expressing EL4 leukemia, followed by control diluent or C20:2 IP on days +1 and +4. Relapse by Kaplan-Meier analysis (allo groups, n = 24 each; TCD group, n = 10). Combined data from 3 identical experiments. *P = .019 for C20:2 allo vs control allo. (D) Representative bioluminescence images of recipients shown in panel C at day 14 after transplant from 2 separate experiments. (E) Irradiated BALB/c mice received whole or TCD spleen from G-CSF–mobilized B10.D2 donors, followed by control diluent, C20:2, or KRN7000 IP on days +1 and +4. Overall survival by Kaplan-Meier analysis and mean (± SE) of clinical scores in surviving mice at the end of each experiment (allo groups, n = 21 each; TCD group, n = 13). Combined data from 3 identical experiments. **P < .02 for KRN7000 allo vs control and C20:2 allo. *P < .05 for C20:2 allo clinical scores vs control allo. (F) Donor T-cell engraftment (Ly9.1neg) at day 50 in transplant recipients from panel E. n = 4-5 in allo groups; n = 3 in the TCD group.

KRN7000 induces hyperacute GVHD in multiple models. (A) Irradiated B6D2F1 received whole spleen from G-CSF–mobilized syngeneic B6D2F1 or allogeneic B6 donors, followed by control diluent, C20:2 at 2 μg or 5 μg per mouse, or KRN7000 at 0.5 μg or 2 μg per mouse IP on days +1 and +4. Overall survival by Kaplan-Meier analysis of combined data from 2 experiments (control allo, n = 14; C20:2 2 μg allo, n = 12; C20:2 5 μg allo, n = 16; KRN7000 2 μg allo, n = 10; KRN7000 0.5 μg allo, n = 16; KRN7000 syn, n = 8). **P < .001 for KRN7000 2 μg allo vs C20:2 2 μg and 5 μg allo. *P < .04 for KRN7000 0.5 μg allo vs C20:2 2 μg and 5 μg allo. (B) Irradiated B6 mice received whole or TCD spleen from G-CSF–mobilized BALB/c donors, followed by control diluent, C20:2, or KRN7000 IP on days +1 and +4. Overall survival by Kaplan-Meier analysis and mean ± SE of clinical scores in surviving mice at the end of each experiment (allo groups, n = 20 each; TCD group, n = 12). Combined data from 3 identical experiments. **P < .02 for C20:2 allo vs control and KRN7000 allo. *P < .05 for control allo clinical scores vs C20:2 and KRN7000 allo. (C) Leukemic relapse in irradiated B6 mice receiving whole or TCD spleen from G-CSF–mobilized BALB/c donors with the addition of host-type luciferase expressing EL4 leukemia, followed by control diluent or C20:2 IP on days +1 and +4. Relapse by Kaplan-Meier analysis (allo groups, n = 24 each; TCD group, n = 10). Combined data from 3 identical experiments. *P = .019 for C20:2 allo vs control allo. (D) Representative bioluminescence images of recipients shown in panel C at day 14 after transplant from 2 separate experiments. (E) Irradiated BALB/c mice received whole or TCD spleen from G-CSF–mobilized B10.D2 donors, followed by control diluent, C20:2, or KRN7000 IP on days +1 and +4. Overall survival by Kaplan-Meier analysis and mean (± SE) of clinical scores in surviving mice at the end of each experiment (allo groups, n = 21 each; TCD group, n = 13). Combined data from 3 identical experiments. **P < .02 for KRN7000 allo vs control and C20:2 allo. *P < .05 for C20:2 allo clinical scores vs control allo. (F) Donor T-cell engraftment (Ly9.1neg) at day 50 in transplant recipients from panel E. n = 4-5 in allo groups; n = 3 in the TCD group.

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