Polyreactive Antibodies Developing Amidst Humoral Rejection of Human Kidney Grafts Bind Apoptotic Cells and Activate Complement
Article first published online: 6 AUG 2013
© Copyright 2013 The American Society of Transplantation and the American Society of Transplant Surgeons
American Journal of Transplantation
Volume 13, Issue 10, pages 2590–2600, October 2013
How to Cite
Porcheray, F., Fraser, J. W., Gao, B., McColl, A., DeVito, J., Dargon, I., Helou, Y., Wong, W., Girouard, T. C., Saidman, S. L., Colvin, R. B., Palmisano, A., Maggiore, U., Vaglio, A., Smith, R. N. and Zorn, E. (2013), Polyreactive Antibodies Developing Amidst Humoral Rejection of Human Kidney Grafts Bind Apoptotic Cells and Activate Complement. American Journal of Transplantation, 13: 2590–2600. doi: 10.1111/ajt.12394
- Issue published online: 24 SEP 2013
- Article first published online: 6 AUG 2013
- Manuscript Accepted: 20 JUN 2013
- Manuscript Revised: 5 JUN 2013
- Manuscript Received: 14 JAN 2013
- Fahd and Nadia Alireza's Research Fund
- Roche Organ Transplantation Research Foundation
- National Institute of Health, National Institute of Diabetes, Digestive and Kidney Diseases. Grant Number: DK083352
Additional supporting information may be found in the online version of this article at the publisher's web-site.
Figure S1: (A) The reactivity of the supernatants from the 107 immortalized B cell clones to apoptotic cells was assessed by flow cytometry using an anti IgG/M FITC-conjugated secondary antibody. The background reactivity of the secondary antibody tovapoptotic cells is also reported (upper left corner histogram). Supernatants considered positive are boxed. (B) Staining of viable jurkat cells with a commercially available anti-human class I antibody (clone w6/32).
Figure S2: Correlation between polyreactivity and reactivity to apoptotic cells. Cumulative reactivity index to dsDNA, Insulin and lysate of HEK293 cells (x-axis) for all 107 B cell antibodies are plotted with reactivity index to apoptotic cells for the same clones (y-axis). Statistical analysis is based on a two-tailed nonparametric spearman's test.
Figure S3: Polyreactive antibody reactivity to phosphatidylserine (PS) and lysophosphatidylcholine (LPC). Cell culture supernatants from the six polyreactive clones as well as from the nonpolyreactive clone 3D4, were tested by ELISA for their reactivity to PS (A) or LPC (B).
Figure S4: Cause of end stage renal failure (ESRF) and serum reactivity to apoptotic cells (A), purified IgG reactivity to apoptotic cells (B) and C4d binding after purified IgG opsonization of apoptotic cells (C). Values obtained with samples from patients who developed ESRF because of autoimmune diseases are labeled in red.
Figure S5: Correlation between sampling time post transplantation and serum IgG reactivity to apoptotic cells (upper), purified IgG reactivity to apoptotic cells (middle) and C4d binding to apoptotic cells (lower). Left panels: Correlation for non-AMR samples. Right panels: Correlation for AMR samples.
Figure S6: Increase in IgG serum reactivity to apoptotic cells at time of AMR. Available samples from five kidney transplant recipients collected before and at time of AMR were assessed for IgG reactivity to apoptotic jurkat cells (upper panel). Variations in serum IgG concentration were measured for the same samples at the same time points (lower panel).
Table S1: Causes of ESRF
Table S2: HLA antibodies status of the patients
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