jkstn The Journal of the Korean Society for Transplantation J Korean Soc Transplant 1598-1711 The Korean Society for Transplantation 10.4285/jkstn.2010.24.4.298 jkstn-24-298 Original Article Expression of E-cadherin, Heat Shock Protein 47, Transforming Growth Factor β1 and C4d in Chronic Allograft Nephropathy KiWon ChunM.D. 1 JunHo ParkM.D. 1 JinCheol JungM.D. 1 DooJin KimM.D. 1 SungGil ParkM.D. 1 JooSeop KimM.D. 1 EunSuk NamM.D. 2 SukJa HyunR.N. 3 LeeSamuelM.D. 1 Departments of Surgery, Kang-dong Sacred Heart Hospital, Hallym University College of Medicine, Seoul, Korea Pathology, Kang-dong Sacred Heart Hospital, Hallym University College of Medicine, Seoul, Korea Transplantation Center, Kang-dong Sacred Heart Hospital, Hallym University College of Medicine, Seoul, Korea 책임저자：이삼열, 서울시 강동구 성내길 150한림대학교 강동성심병원 외과, 134-010 Tel: 02-2224-2222, Fax: 02-2224-2570 E-mail: slee@hallym.or.kr 01 2010 10 12 2010 24 4 298 305 Copyright © 2010 The Korean Society for Transplantation 2010 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract Background

Chronic allograft nephropathy (CAN), which causes graft failure, is related to tubular atrophy and interstitial fibrosis. E-cadherin is a well-known epithelial marker and heat shock protein (HSP)-47 is a collagen-specific molecular chaperone that regulates collagen synthesis. Transforming growth factor (TGF)-β1, a profibrotic cytokine, downregulates E-cadherin and induces expression of mesenchymal markers in an in vitro model. C4d expression is considered a poor prognostic marker for graft survival. This study evaluated the relationship between the expression of E-cadherin, HSP47, TGF-β1, and C4d with the prognosis for CAN.

 Methods

Between March 1991 and August 2007, we performed renal allograft biopsies on 42 recipients with deteriorating renal function. CAN was diagnosed according to the chronic allograft damage index (Banff classification). Renal allograft biopsies were examined for the expression of E-cadherin, HSP47, TGF-β1, or C4d by immunohistochemistry. The HSP47, TGF-β1, and E-cadherin staining was scored semiquantitatively by analyzing ten different fields of cortical interstitium and tubules. Biopsies with endothelial C4d staining in peritubular capillaries (≥25%) were designated as C4d-positive.

 Results

Of 42 recipients, 17 (40.5%) were in the graft survival group (GS) and 25 (59.5%) were in the graft failure group (GF). E-cadherin expression in tubular cells of the GS was much higher than that of the GF (94.1% vs 52%, P=0.04). HSP47 expression in tubular cells and interstitium in the GF was much higher than that in the GS (84% vs 35.3%, P=0.001). TGF-β1 expression in tubular cells and interstitium in the GF was much higher than that in the GS (72% vs 23.5%, P=0.02).

 Conclusions

E-cadherin, HSP47, and TGF-β1 expression was strongly correlated with the CAN prognosis.

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E-cadherin expression in Implantation biopsy vs Functioning group (CAN) vs non functioning group (CAN), arrow (high expression), arrow head (low expression) (×100). (A) Implantation biopsy, high expression of E-cadherin. (B) CAN, Functioning group mild loss of expression of E-cadherin. (C) CAN, non functioning group entire loss of expression. Abbreviation: CAN, chronic allograft nephropathy.

HSP47 expression in Implantation biopsy vs Functioning group (CAN) vs non functioning group (CAN), arrow (high expression), arrow head (low expression) (×100). (A) Implantation biopsy, no expression of HSP47. (B) CAN, functioning group, low expression in tubule. (C) CAN, non functioning group, high expression in tubule and interstitium. Abbreviations: HSP, heat shock protein; CAN, chronic allograft nephropathy.

TGF-ß1 expression in implantation biopsy vs functioning group (CAN) vs non functioning group (CAN), arrow (high expression), arrow head (low expression) (×100). (A) Implantation biopsy, no expression of TGF-ß1. (B) CAN, Functioning group mild expression in interstitium. (C) CAN, non functioning group high expression in tubule & inerstitium. Abbreviations: TGF, transforming growth factor; CAN, chronic allograft nephropathy.

Correlation of E-cadherin E-cadherin, HSP-47, TGF-β1, C4d expression and graft survivals after transplantation. (A) E-cadherin.(B) HSP-47. (C) TGF-b1. (D) C4d. Abbreviations: HSP, heat shock protein; TGF, transforming growth factor.

Banff 97 diagnostic categories for renal allograft biopsies-update

chronic/sclerosing allograft nephropathy Fibrosing changes in the allograft with or without features of true alloimmune injury to the graft may coincide with categories 2, 3 and 4.
Grade Histologic finding
Grade I (mild) Mild interstitial fibrosis and tubular atrophy without (a) or with (b) specific changes suggesting chronic rejection
Grade II (moderate) Moderate interstitial fibrosis and tubular atrophy (a) or (b)
Grade III (severe) Severe interstitial fibrosis and tubular atrophy and tubular loss (a) or (b)

Patients characteristics in post-transplant chronic allograft nephropathy

Variables n=42
Gender (male/female) 28/14
Donor (living/deceased) 28/14
Recipient Age (year) 41.6±10.2
Donor Age (year) 35.7±11.9
Immunosuppressant (FK/CyA) 30/12
HLA mismatch (A, B, DR) 2.3±0.9
Timing of biopsy post KT (month) 66.6±47.2
Allograft rejection episode (%) 19.0 (8/42)
Creatinine at biopsy (mmol/L) 2.85±1.4
Significant proteinuria (＞0.5 g/day) 12/30
Graft survival rate (%) 40.5 (17/42)

Abbreviations: CyA, cyclosporin A; HLA, human leukocyte anti-gen; KT, kidney transplantation.

Correlation between Banff score and functioning graft/ graft loss

Banff score Biopsy P
Functioning graft (n=17) Graft loss (n=25)
Chronic allograft 6 11 0.573
　 glomerulopathy (CG)      
Chronic interstitial fibrosis 4 12 0.107
　 (CI)      
Chronic tubular atrophy 4 18 0.02
　 (CT)      
Chronic vascular fibrous 5 15 0.051
intimal thickening (CV)      

Correlation between markers and proteinuria, serum creatinine

Marker Proteinuria P Cr a P
(+) (−)
E-cadherin (+) 11 2 0.96 2.75 0.40
  (−) 20 9   3.04  
HSP47 (+) 11 16 0.019 3.05 0.477
  (−) 1 14   2.46  
TGF-ß1 (+) 5 20 0.136 3.10 0.380
  (−) 7 10   2.40  
C4d (+) 3 9 0.746 4.14 ＜0.05
  (−) 9 21   2.37  

Abbreviations: HSP, heat shock protein; TGF, transforming growth factor.

Value at biopsy.