Figure 1
Figure 1. Vitamin A metabolism is upregulated during acute GVHD. (A) Lethally irradiated B10.BR recipients were injected with 107 T-cell–depleted (TCD) BM cells and 1.5 × 107 splenocytes from fully MHC-mismatched B6 mice. On days 7 and 14 after BMT, tissues from lung, liver, small intestine, and colon were harvested and analyzed using a B16-DR5 assay (n = 4/group). The fold change was calculated by dividing the value of the non-GVHD group or GVHD group by the value of naive mouse group. (B) Lethally irradiated B10.BR recipients injected with 107 wt B6 TCD-BM cells and 3 × 106 purified RARE-luc B6 T cells. On days 7 and 21 after BMT, tissues from liver, small intestine, and colon were harvested and analyzed using a LC-(MS)-MS assay (n = 4/group). (C) Lethally irradiated B10.BR recipients were injected with 107 BM cells and 1.5 × 107 B6 splenocytes from wt B6 mice. Recipient mice (n = 4/group) were sacrificed on day 14 along with 4 naive B10.BR control mice, and sections from frozen tissue blocks were analyzed for expression of CD11c+ (green), CD45+ (blue), and either retinaldehyde dehydrogenase 1 (RALDH1) (red) or RALDH2 (red). The boxed area in (C) is indicated at higher magnification in (C′, C″). Arrows points out triple labeling (CD11c/CD45/RALDH1 or CD11c/CD45/RALDH2). Data shown are representative of 4 mice/group. Fluorescence was detected using an Olympus FluoView 1000 BX2 Upright confocal laser scanning microscope. (Original magnification ×400.) (D) Lethally irradiated B6-Ly5.2/Cr or B10.BR recipients were injected with 107 BM cells and 1.5 × 107 splenocytes from fully MHC-mismatched B6 mice. Lamina propria lymphocytes from the small intestine and lymphocytes from the MLNs were isolated on day 14 and donor (CD45.1− or H2kb+) CD11c+ and CD11b+ cells were evaluated for ALDH enzymatic activity by measuring the Aldefluor mean fluorescence intensity (MFI) (n = 4/group). (E) Lethally irradiated B6 or B10.BR recipients were transplanted with 107 BM cells and 3 × 106 purified RARE-luc B6 T cells. RARE-luc T cells were quantified by emitted photons over the total body area and within individual organs at serial time points after BMT (n = 3-4/group). *P > .05; **P > .01; and ***P > .001.

Vitamin A metabolism is upregulated during acute GVHD. (A) Lethally irradiated B10.BR recipients were injected with 107 T-cell–depleted (TCD) BM cells and 1.5 × 107 splenocytes from fully MHC-mismatched B6 mice. On days 7 and 14 after BMT, tissues from lung, liver, small intestine, and colon were harvested and analyzed using a B16-DR5 assay (n = 4/group). The fold change was calculated by dividing the value of the non-GVHD group or GVHD group by the value of naive mouse group. (B) Lethally irradiated B10.BR recipients injected with 107 wt B6 TCD-BM cells and 3 × 106 purified RARE-luc B6 T cells. On days 7 and 21 after BMT, tissues from liver, small intestine, and colon were harvested and analyzed using a LC-(MS)-MS assay (n = 4/group). (C) Lethally irradiated B10.BR recipients were injected with 107 BM cells and 1.5 × 107 B6 splenocytes from wt B6 mice. Recipient mice (n = 4/group) were sacrificed on day 14 along with 4 naive B10.BR control mice, and sections from frozen tissue blocks were analyzed for expression of CD11c+ (green), CD45+ (blue), and either retinaldehyde dehydrogenase 1 (RALDH1) (red) or RALDH2 (red). The boxed area in (C) is indicated at higher magnification in (C′, C″). Arrows points out triple labeling (CD11c/CD45/RALDH1 or CD11c/CD45/RALDH2). Data shown are representative of 4 mice/group. Fluorescence was detected using an Olympus FluoView 1000 BX2 Upright confocal laser scanning microscope. (Original magnification ×400.) (D) Lethally irradiated B6-Ly5.2/Cr or B10.BR recipients were injected with 107 BM cells and 1.5 × 107 splenocytes from fully MHC-mismatched B6 mice. Lamina propria lymphocytes from the small intestine and lymphocytes from the MLNs were isolated on day 14 and donor (CD45.1− or H2kb+) CD11c+ and CD11b+ cells were evaluated for ALDH enzymatic activity by measuring the Aldefluor mean fluorescence intensity (MFI) (n = 4/group). (E) Lethally irradiated B6 or B10.BR recipients were transplanted with 107 BM cells and 3 × 106 purified RARE-luc B6 T cells. RARE-luc T cells were quantified by emitted photons over the total body area and within individual organs at serial time points after BMT (n = 3-4/group). *P > .05; **P > .01; and ***P > .001.

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