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. 2008 Mar 17;105(12):4796–4801. doi: 10.1073/pnas.0712051105

Fig. 3.

Fig. 3.

SAHA augmented induction of chimerism in old BALB/c mice. Old (>16 weeks) BALB/c mice were conditioned with anti-CD3 alone or anti-CD3 and SAHA and transplanted with C57BL/6 donor BM (2 × 106) and CD4+ T-depleted spleen cells (4 × 106). The recipients were monitored for clinical signs of GVHD daily and body weight weekly. (A) At 8 weeks after HCT, blood mononuclear cells were stained for H-2b (donor marker), TCRαβ, B220, and Mac-1/Gr-1. The percentages of donor T, B, and macrophage cells are shown. One representative sample of 12 mice in each group is shown. (B) Body weight change curves of HCT recipients conditioned with anti-CD3 alone (nonchimeric) or anti-CD3 plus SAHA (chimeric). There were 12 mice in each group. Mean ± SE is shown at each time point. (C) Yield of residual DAPI− live T cells in spleen of mice conditioned with anti-CD3 or anti-CD3 plus SAHA 9 days after anti-CD3 injection at the day of HCT. (D) Sorted T cells (0.2 × 106) from the spleen of conditioned BALB/c mice 9 days after anti-CD3 injection were stimulated with C57BL/6 dendritic cells (0.1 × 106) for 5 days. The proliferation was measured with 3H-TdR incorporation, and the stimulation index was calculated by formula: {[cpm of responder × stimulator] − [cpm of responder alone]} ÷ [cpm of responder alone]. Mean ± SE of four mice in each group is shown. (E) At 5 days after HCT, donor-type spleen cells from congenic C57BL/6 (CD45.1) and CFSE-labeled host-type spleen cells from BALB/c were injected into recipients conditioned with anti-CD3 alone or anti-CD3 plus SAHA. Then, 18 h later, spleen cells were harvested and stained with anti-CD45.1. Staining is shown in CD45.1 versus CFSE. One representative FACS pattern of four mice in each group is shown. The mean ± SE of CD45.1+ or CFSE+ cells in recipients conditioned with anti-CD3 alone versus recipients conditioned with anti-CD3 plus SAHA were 1.21 ± 0.07% versus 1.80 ± 0.08% or 0.35 ± 0.03% versus 0.20 ± 0.01%. (F) The ratio of residual CD45.1+ donor-type cells versus CFSE+ host-type cells were calculated, and mean ± SE of 4 recipients in each group is shown.