Article first published online: 14 OCT 2012
Copyright © 2012 American Association for the Study of Liver Diseases
Volume 56, Issue 5, pages 1924–1933, November 2012
How to Cite
Höchst, B., Schildberg, F. A., Böttcher, J., Metzger, C., Huss, S., Türler, A., Overhaus, M., Knoblich, A., Schneider, B., Pantelis, D., Kurts, C., Kalff, J. C., Knolle, P. and Diehl, L. (2012), Liver sinusoidal endothelial cells contribute to CD8 T cell tolerance toward circulating carcinoembryonic antigen in mice. Hepatology, 56: 1924–1933. doi: 10.1002/hep.25844
Potential conflict of interest: Nothing to report.
Supported by a grant from the German Cancer Aid (to P. K. and L. D.).
- Issue published online: 31 OCT 2012
- Article first published online: 14 OCT 2012
- Accepted manuscript online: 18 MAY 2012 11:49AM EST
- Manuscript Accepted: 8 MAY 2012
- Manuscript Received: 8 DEC 2011
Additional Supporting Information may be found in the online version of this article.
|HEP_25844_sm_SuppFig1.tif||1100K||Supporting Information Figure 1: Uptake of circulating CEA and OVA in antigen-presenting cells in the spleen. Alexa647-labelled CEA (A) or OVA (B) protein was injected intravenously into C57BL/6 and MR-/- mice. 5 minutes after injection splenocytes were isolated stained for CD11b or CD11c and analyzed by flow cytometry. Mean fluorescence intensity of splenic CD11c+ or CD11b+ from C57BL/6 or MR-/- mice previously injected with CEAAlexa647 (A) or OVAAlexa647 (B). Shown are cumulative results from 3 independent experiments.|
|HEP_25844_sm_SuppFig2.tif||1128K||Supporting Information Figure 2: CEA does not affect antigen presenting cells. (A) LSEC from C57BL/6 or TLR4-/- mice were stimulated with LPS (0.1μg/ml) or with indicated amount of CEA or OVA in vitro. IL-6 secretion was measured by ELISA. Shown are cumulative results from 3 independent experiments.|
|HEP_25844_sm_SuppFig3.tif||1287K||Supporting Information Figure 3: Growth of MC38 tumors not expressing CEA is not affected by CEA-specific T cells. C57BL/6 (A), B7H1-deficient (B) or MR-deficient (C) mice received 2 mg of CEA protein i.v. every second day, were immunized with the pGT64 CEA plasmid or were left untreated. After 2 weeks splenic CD8 T cells from these C57BL/6 (A) B7H1-/- (B) or MR-/- mice (C) were obtained and 5x106 total CD8 T cells were adoptively transferred into Rag2-deficient mice. Three days later, 5x106 MC-38 cells were injected into the left flank. Graphs show tumor growth of MC38 in Rag2-/- mice that received CD8 T cells from C57BL/6 (A), B7H1-/- (B) or MR-/-mice (C).|
|HEP_25844_sm_SuppFig4.tif||769K||Supporting Information Figure 4: Phenotypical tolerant CD8 T cells do not proliferate or degranulate upon stimulation. PBMC from healthy individuals were isolated from blood and sorted for CD8, CD45RO, CD62L and CD25 using FACS-Sort. Isolated T cells labeled with CFSE (A, B) or not (C) and were stimulated with anti-CD3ε/CD28 beads(A-C). Proliferation was measured by CFSE dilution using flow cytometry. A: % of proliferated CD8 T cells. B: CFSE dilution of sorted CFSE-labeled CD8 T cells. C: Surface CD107a expression on sorted CD8 T cells 1h after stimulation. Shown are results from at least 3 independent experiments.|
|HEP_25844_sm_SuppTable1.doc||30K||Supporting Information Table 1.|
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