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NDT Advance Access published online on October 25, 2005
Nephrology Dialysis Transplantation, doi:10.1093/ndt/gfi229
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© The Author [2005]. Published by Oxford University Press on behalf of ERA-EDTA. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org
Received July 13, 2005
Accepted September 27, 2005

Original Articles

Glomerular crescent-related biomarkers in a murine model of chronic graft-versus-host disease

Shuk-Man Ka 1, Abdalla Rifai 2, Jan-Hen Chen 3, Chao-Wen Cheng 1, Hao-Ai Shui 4, Herng-Sheng Lee 5, Yuh-Feng Lin 6, Lai-Fa Hsu 5, and Ann Chen 5*

1 Graduate Institute of Life Sciences, Taiwan, ROC
2 Department of Pathology, Rhode Island Hospital, RI, USA
3 Biochip R&D Center, Taiwan, ROC; School of Density, Taiwan, ROC
4 Graduate Institute of Medical Sciences, National Defense Medical Center, Taiwan, ROC
5 Department of Pathology, Taiwan, ROC
6 Department of Internal Medicine, Tri-Service General Hospital, Taiwan, ROC

* To whom correspondence should be addressed.
Ann Chen, E-mail: doc31717{at}ndmctsgh.edu.tw



  Abstract

Background. We examined the alterations in gene expression associated with the development of crescentic glomerulonephritis in murine chronic graft-versus-host disease, a model for human systemic lupus erythematosus.

Methods. The disease was induced in (C57BL/6 x DBA/2) F1 hybrids by injection of DBA/2 lymphocytes leading to deposition of auto-antibodies in the glomeruli, and a lupus type of nephritis morphologically. After extensive crescent formation at week 9 of disease, cDNA microarray analysis was performed and highly expressed genes were evaluated as molecular markers by real-time reverse transcription-polymerase chain reaction (RT-PCR), in situ hybridization, immunohistochemistry and immunoassay of urine proteins.

Results. Six genes, secreted acidic cysteine-rich glycoprotein (Sparc), thymosin beta 10 (Tmsb10), S100 calcium-binding protein A6 (S100a6), annexin A2 (Anxa2), osteopontin (OPN) and lipocalin 2 (Lcn2), were quantified by real-time RT-PCR in laser microdissected glomeruli in a time course manner. Sparc was detected early before the onset of proteinuria and continued to increase throughout the course of the disease. The expression of Tmsb10, S100a6 and Anxa2 coincided with heavy proteinuria. By week 9, OPN and Lcn2 were highly expressed. The expression of proteins encoded by these genes was predominant in the glomerular crescent. The protein levels of Sparc, OPN and Lcn2 in urine were significantly elevated.

Conclusions. These findings implicate these six genes in the development of glomerular crescents. More importantly, detection of Sparc, OPN and Lcn2 in urine may mean that these molecules could serve as important biomarkers for non-invasive diagnosis of glomerular crescents.

Keywords: Anxa2; crescentic lupus nephritis; laser microdissection; S100a6; Sparc; Tmsb10.
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