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Blood, Vol. 113, Issue 10, 2352-2362, March 5, 2009
Interleukin-23 secretion by donor antigen-presenting cells is critical for organ-specific pathology in graft-versus-host disease
Blood Das et al.
113: 2352
Supplemental materials for: Das et al
Files in this Data Supplement:
- Figure S1. GVHD protection associated with transplantation of IL-23−∕− marrow grafts is not murine model-dependent (JPG, 58.9 KB)
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(A). Lethally irradiated FVB mice (1000 cGy) were transplanted with either BM (10 × 106) alone from B6 (■, n=8) or IL-23−∕− (≤, n=5) animals or BM and spleen cells from B6 (●, n=19) or IL-23−∕− mice (○, n=17). Spleen cell inoculums were adjusted so that recipient mice received the same dose of T cells (0.4–0.6 × 106) from either donor. The percent weight change over time of mice from all four groups is depicted. Data are cumulative results from four experiments. (B–E). Spleen cellularity (B), total number of splenic B cells (C), thymus size (D) and absolute number of double positive (CD4+CD8+) thymocytes (E) in mice transplanted with B6 BM alone (n=4, hatched bars), IL-23−∕− BM alone (n=3, gray bars), B6 BM plus spleen cells (n=14, black bars) or IL-23−∕− BM and spleen cells (n=13, white bars). Data are presented as the mean ± SEM. Statistics: * p ≤ 0.05, ** p < 0.01.
- Figure S2. Protection from GVHD is due to absence of donor APC-derived IL-23 secretion (JPG, 13.9 KB)
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Lethally irradiated Balb/c (900 cGy) mice were transplanted with B6 (n=14, black bars) or IL-23−∕− BM (n=15, white bars) (10 × 106) plus purified B6 T cells at a CD4:CD8 ratio of 2:1 (total T cell dose 1.75–2.25 × 105) from B6 animals. Mice were sacrificed 29 days post transplantation and GVHD target tissues (colon, liver and lung) were examined for pathological damage using a semiquantitative scoring system as detailed in Methods. Data are presented as the mean ± SEM and are the cumulative results from three independent experiments. Statistics: ** p ≤ 0.01.
- Figure S3. Attenuation of proinflammatory cytokine production in the colon after transplantation of IL-23−∕− marrow grafts is not a model-dependent phenomenon (JPG, 32.2 KB)
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Lethally irradiated (1000 cGy) FVB mice were transplanted with B6 BM (10 × 106) plus spleen cells (1.5–2 × 106) (n=18, black bars) or IL-23−∕− BM and spleen cells adjusted to yield an equivalent number of T cells (n=18, white bars). Mice were sacrificed 21–29 days post transplantation. Colon tissue from individual mice was cultured overnight in medium as described in Methods and tissue supernatants were assayed for levels of the indicated proinflammatory cytokines by multiplex. Data are presented as mean amount of cytokine divided by the weight of cultured colon tissue ± SEM and are cumulative results from two experiments. Statistics: * p ≤ 0.05, ** p < 0.01.
- Figure S4. Donor-derived CD4+ T cells with an activated/memory phenotype are present in the colon during GVHD (JPG, 53.9 KB)
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(A). Lethally irradiated (900 cGy) Balb/c mice were transplanted with B6 BM (10 × 106) and spleen cells (0.4–0.6 × 106) (black bars) or IL-23−∕− BM (10 × 106) and spleen cells adjusted to yield the same T cell dose (white bars). Mice in each cohort were sacrificed 28–30 days post transplantation. Lymphocytes were isolated from the spleen, liver, or colon of mice (n=4–5/group per experiment) and the absolute number of CD4+ T cells was quantitated. Data are presented as the mean ± SEM and are cumulative results from 4 independent experiments. (B). Representative dot plots demonstrating CD44 and CCR7 expression on gated CD4+ T cells derived from spleen, liver and colon of GVHD animals transplanted with B6 BM and spleen cells 4 weeks post transplantation. Statistics: * p ≤ 0.05.
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