Figure 6.
Signaling through the CB2R regulates the severity of chronic GVHD. (A-B) The eGFP expression percentage on macrophages derived from the liver, lung, or spleen of normal CB2ReGFP mice (n = 20-23) is shown. Representative dot plots showing eGFP expression on macrophages from these tissue sites is shown in panel B. (C-D) Lethally irradiated Balb/c mice received transplants of B10.D2 BM alone (n = 9) or together with 15 × 106 B10.D2 spleen cells. Animals that received spleen cells were treated with the CB2R antagonist SR144528 (3 mg/kg; n = 24) or a vehicle control (n = 22) for 14 days beginning on the day of transplantation. Cumulative pathological chronic GVHD skin scores of mice 30 to 40 days posttransplantation are depicted in panel C. Representative hematoxylin and eosin–stained sections of the skin on day 35 are shown in panel D. Original magnification is ×100 for photomicrographs. Data are derived from 5 experiments. (E) Immunohistochemical staining depicting CD3 and F4/80 positive cells (brown coloration) in the skin of mice treated with SR144528. (F) mRNA expression of transforming growth factor β (TGF-β), galectin 3 (GAL-3), and α smooth muscle (α-SMA) in the skin of animals 30 to 40 days posttransplantation. Results are from 3 experiments, with 6 to 15 mice per group. (G-J) Lethally irradiated (850 cGy) Balb/c mice received transplants of B6 Rag-1−/− BM alone, B6 Rag-1−/− BM and B6 spleen cells (adjusted to yield an αβ T-cell dose of 0.4 × 106), or CB2R−/− BM and spleen cells (adjusted to yield the same αβ T-cell dose). Representative hematoxylin and eosin– and trichrome-stained sections of the skin on day 45 are shown in panels G and H, respectively. Original magnification is ×100 for photomicrographs. (I) Pathology scores of the skin 40 to 45 days posttransplantation in chronic GVHD mice. Results are from 3 experiments, with 7 to 13 mice per group. (J) mRNA expression of TGF-β, GAL-3, and α-SMA in the skin of animals (n = 8-15 mice per group) 40 to 45 days posttransplantation. Results are from 3 experiments. Data in panels in A, C, F, I, and J are presented as the mean ± standard deviation. *P < .05, **P < .01, ***P < .001, ****P < .0001.

Signaling through the CB2R regulates the severity of chronic GVHD. (A-B) The eGFP expression percentage on macrophages derived from the liver, lung, or spleen of normal CB2ReGFP mice (n = 20-23) is shown. Representative dot plots showing eGFP expression on macrophages from these tissue sites is shown in panel B. (C-D) Lethally irradiated Balb/c mice received transplants of B10.D2 BM alone (n = 9) or together with 15 × 106 B10.D2 spleen cells. Animals that received spleen cells were treated with the CB2R antagonist SR144528 (3 mg/kg; n = 24) or a vehicle control (n = 22) for 14 days beginning on the day of transplantation. Cumulative pathological chronic GVHD skin scores of mice 30 to 40 days posttransplantation are depicted in panel C. Representative hematoxylin and eosin–stained sections of the skin on day 35 are shown in panel D. Original magnification is ×100 for photomicrographs. Data are derived from 5 experiments. (E) Immunohistochemical staining depicting CD3 and F4/80 positive cells (brown coloration) in the skin of mice treated with SR144528. (F) mRNA expression of transforming growth factor β (TGF-β), galectin 3 (GAL-3), and α smooth muscle (α-SMA) in the skin of animals 30 to 40 days posttransplantation. Results are from 3 experiments, with 6 to 15 mice per group. (G-J) Lethally irradiated (850 cGy) Balb/c mice received transplants of B6 Rag-1−/− BM alone, B6 Rag-1−/− BM and B6 spleen cells (adjusted to yield an αβ T-cell dose of 0.4 × 106), or CB2R−/− BM and spleen cells (adjusted to yield the same αβ T-cell dose). Representative hematoxylin and eosin– and trichrome-stained sections of the skin on day 45 are shown in panels G and H, respectively. Original magnification is ×100 for photomicrographs. (I) Pathology scores of the skin 40 to 45 days posttransplantation in chronic GVHD mice. Results are from 3 experiments, with 7 to 13 mice per group. (J) mRNA expression of TGF-β, GAL-3, and α-SMA in the skin of animals (n = 8-15 mice per group) 40 to 45 days posttransplantation. Results are from 3 experiments. Data in panels in A, C, F, I, and J are presented as the mean ± standard deviation. *P < .05, **P < .01, ***P < .001, ****P < .0001.

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