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Blood, Vol. 109, Issue 9, 3830-3838, May 1, 2007
Effective graft depletion of MiHAg T-cell specificities and consequences for graft-versus-host disease
Blood de Witte et al.
109: 3830
Supplemental materials for: de Witte et al, Vol 109, Issue 9, 3830-3838
Files in this Data Supplement:
- Figure S1. Comparative analysis of H60-specific T-cell responses by intracellular IFNγ staining or Kb-H60 tetramer staining (PDF, 124 KB) -
Splenocytes from female B6 mice were either depleted with Kb-H4 and Kb-H60 tetramers or mock depleted. Subsequently, 25 × 106 T cells plus 5 × 106 T-cell–depleted bone marrow cells were transferred into lethally irradiated male BALB.B recipients (n = 6 in both groups). Blood was sampled at day 10 after HSCT, and antigen-specific CD8+ T-cell responses were determined ex vivo by intracellular IFN staining (top panels in panel A; circles in panel B) or by staining with H60-tetramers for each individual mouse (bottom panels in panel A; diamonds in panel B).
- Figure S2. Nonlinear expansion of antigen-specific T cells during in vitro restimulation (PDF, 68.1 KB) -
T cells (25 × 106) plus 5 × 106 T-cell–depleted bone marrow cells were transferred into lethally irradiated male BALB.B recipients. Splenocytes were harvested at day 24 after transplantation, after which the amount of H60-specific T cells was determined to be 27%, by intracellular IFN staining following a 5-hour stimulation with H60 peptide. These B6 → BALB.B splenocytes were mixed with splenocytes from naive B6 mice, and the cells were restimulated in vitro for 14 days with CD8+ T-cell–depleted irradiated male BALB.B target splenocytes. The resulting cell cultures were tested for reactivity against H60 by intracellular IFN staining following a 5-hour stimulation with H60 peptide. The x-axis represents the percentage of H60-specific T cells present at the start of culture (day 0); the y-axis represents the percentage of H60-specific T cells present after in vitro restimulation (day 14).
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