For GVHD models, mice were irradiated with 12 Gy (BDF1) or 10 Gy (B6) twelve hours prior to adoptive transfer of MACS isolated splenic and LN pan T cells (Miltenyi Biotec) and TCD-BM (MACS thy1.2-microbeads as previously described (22 (link)). For the B6 to BDF1 GVHD/CTL mice were not irradiated and 2.5 × 107 total splenocytes/mouse were adoptively transferred. For CD25 depletion, pan T cells were isolated with CD25-biotin (BD biosciences) and anti-biotin microbeads (Miltenyi), and passed over LD depletion columns (Miltenyi). 200 ug/mouse of MH5A or control Ab was injected via tail vein, and mice were monitored daily for all experiments.
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Adoptive T Cell Transfer GVHD Model
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Other organizations : Yale University
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Variable analysis
independent variables
- Radiation dose (12 Gy for BDF1 mice, 10 Gy for B6 mice)
- Adoptive transfer of MACS isolated splenic and LN pan T cells and TCD-BM
- Depletion of CD25+ cells from pan T cells
- Injection of MH5A antibody or control antibody
- Survival and health monitoring of mice
- Time between irradiation and adoptive transfer (12 hours)
- Cell types used for adoptive transfer (MACS isolated splenic and LN pan T cells, TCD-BM)
- Method of CD25+ cell depletion (CD25-biotin and anti-biotin microbeads, LD depletion columns)
- Dose of antibody injection (200 ug/mouse)
- Positive control: B6 to BDF1 GVHD/CTL mice without irradiation and 2.5 × 10^7 total splenocytes/mouse adoptively transferred
- Negative control: Control antibody injection
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