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Blood, Vol. 111, Issue 5, 2919-2928, March 1, 2008
Prevention of acute graft-versus-host disease by blocking T-cell entry to secondary lymphoid organs
Blood Beilhack et al.
111: 2919
Supplemental materials for: Beilhack et al
Files in this Data Supplement:
- Figure S1. Skin and liver are targets in lymph node and Peyer’s patch deficient B6.LTα−/− but not in splenectomized B6.LTα−/− recipients (JPG, 76.3 KB)
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Upper panel: Immunofluorescence microscopy of myeloablative conditioned recipients on day+6 after aHCT. (A) C57Bl/6 wild type (B6.WT) recipients and (B) C57Bl/6-Lymphotoxin alpha−/− (B6.LT −/−) recipients show infiltration of CD90.1 positive (Alexa-488, green fluorescent) allogeneic donor T lymphocytes (white arrowheads). (C) In contrast, the skin of splenectomized B6.LT−/− recipients (B6.LT−/−SplEx) does not show infiltration by allogeneic donor T lymphocytes. Superficial epidermal layers cause characteristic autofluorescence. Nuclear counterstaining was performed with DAPI (blue, 400×). Lower panel: H&E staining of liver samples on day+6 after aHCT. (D) B6.WT recipients and (E) B6.LT−/− recipients display severe damage of bile ducts (black arrowheads), with loss of epithelia and lymphocyte infiltration, indicating acute liver GVHD (Grade 1-2). (F) In contrast, the liver of B6.LT−/−SplEx recipients shows normal bile ducts (black arrowheads), without lymphocytic infiltration. H&E, 400×.
- Figure S2. Delayed skin infiltration after transfer of in vivo primed alloreactive T-cells into conditioned secondary allogeneic recipients irrespective of the original priming site (JPG, 65.4 KB)
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In contrast to intestinal and hepatic recruitment of alloreactive T cells within 18 hours after secondary transfer, skin infiltration occurred days later. In vivo BLI imaging demonstrates bioluminescence signals from ears depicting skin infiltration by allogeneic luc+ T-cells. Previously we have described delayed skin GVHD target manifestation as compared to intestinal GVHD in primary aHCT recipients, which would be in accordance with delayed skin infiltration in secondary aHCT recipients. However, secondary migration phenomena resulting in skin infiltration cannot be ruled out because days passed from the injection of the in vivo primed cells to their explicit appearance in the skin. Importantly, the delayed skin infiltration by allogeneic luc+ T cells occurred regardless of the initial priming site (mesenteric lymph nodes, peripheral lymph nodes or spleen).
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