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. 2006 Feb 20;203(2):437–447. doi: 10.1084/jem.20051775

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Copyright © 2006, The Rockefeller University Press

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Figure 3. — Organ-specific detection of antigen presentation by ex vivo T cell proliferation assay. Antigen presentation in spleens, peripheral and mesenteric lymph nodes, bone marrow, and livers (unfractionated and fractionated into CD45⁺ and CD45⁻ cells) was assessed independently using an in vitro T cell proliferation assay 4 wk after transplantation. bm8 recipients of liver transplants from [bm8→B6] bone marrow chimeras (hepatic k^b expression, open graphs) were compared with B6 recipients of liver grafts from [B6→B6] bone marrow chimeras (systemic k^b expression, filled graphs). 4 wk after transplantation, transplant recipients were injected with SIINFEKL peptide, and then cell suspensions from spleens, peripheral and mesenteric lymph nodes, bone marrow, and livers were cocultured with CFSE-stained OT-I cells (CD90.1 background). Organ-specific antigen presentation was determined by dilution of CFSE in OT-I cells. The histograms show OT-I T cells based on their expression of CD90.1. These data demonstrate that the capacity to present the SIINFEKL peptide in the hepatic k^b expression group was exclusive to the CD45⁻ and unfractionated intrahepatic cells of the transplanted livers. Data are representative of three independent experiments with two mice per group, and fractionation of liver cells based on CD45 expression was repeated twice with two independently analyzed animals in each group.