[Correction made after online publication December 27, 2012: the author names have been updated.]
Blockade of P-Selectin Is Sufficient to Reduce MHC I Antibody-Elicited Monocyte Recruitment In Vitro and In Vivo
Article first published online: 27 DEC 2012
© Copyright 2012 The American Society of Transplantation and the American Society of Transplant Surgeons
American Journal of Transplantation
Volume 13, Issue 2, pages 299–311, February 2013
How to Cite
Valenzuela, N. M., Hong, L., Shen, X.-D., Gao, F., Young, S. H., Rozengurt, E., Kupiec-Weglinski, J. W., Fishbein, M. C. and Reed, E. F. (2013), Blockade of P-Selectin Is Sufficient to Reduce MHC I Antibody-Elicited Monocyte Recruitment In Vitro and In Vivo. American Journal of Transplantation, 13: 299–311. doi: 10.1111/ajt.12016
- Issue published online: 28 JAN 2013
- Article first published online: 27 DEC 2012
- Manuscript Accepted: 24 OCT 2012
- Manuscript Revised: 19 OCT 2012
- Manuscript Received: 19 JUN 2012
- NIH. Grant Number: U01AI077821
- National Heart, Lung and Blood Institute. Grant Number: RO1 HL 090995
- Ruth L. Kirschstein National Research Service Award. Grant Number: T32HL69766
Figure S1: HLA I antibody binding to human aortic endothelial cells. Human aortic endothelial cells were stained with W6/32 (1 μg/mL). Antibody binding was detected by staining with goat antimouse-FITC secondary antibody. Representative flow cytometric histograms are shown, where the dashed line is unstained control and the dark line is HLA I antibody.
Figure S2: Dose-dependent response of P-selectin induction by HLA I crosslinking. (A) HAEC were treated with HLA I antibody at 1 μg/mL, 5 μg/mL or 10 μg/mL, or isotype control mIgG at 1 μg/mL for 30 min. (B) HAEC were treated with mIgG or HLA I antibody at 5 μg/mL for the indicated times. Cell surface P-selectin was stained with PE-conjugated anti-P-selectin. Graph shows (A) mean fold increase in P-selectin positive cells normalized to mIgG from multiple independent measurements (n=13) or (B) percentage of cells gated positive for P-selectin from one representative experiment out of three. (C) A representative histogram of P-selectin staining on unstimulated (gray fill) or HLA I antibody-activated (bold line) endothelial cells is shown.
Figure S3: Long-term exposure to HLA I antibodies does not alter endothelial expression of E-selectin. HAEC were treated with HLA I antibody or isotype control mIgG (5 μg/mL), or TNFα (10 pg/mL) for the indicated times. Cell surface E-selectin was stained with PE-conjugated anti-E-selectin. Results are expressed as median fluorescence intensity of E-selectin staining from one representative experiment out of two similar.
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