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Volume 60, Issue 3, Pages 1173-1181 (September 2001) Prolonged cold preservation augments vascular injury independent of renal transplant immunogenicity and function Duska Dragun, Uwe Hoff, Joon-Keun Park, Yan Qun, Wolfgang Schneider, Friedrich C. Luft, Hermann Haller Kidney International Volume 60, Issue 3, Pages 1173-1181 (September 2001) DOI: 10.1046/j.1523-1755.2001.0600031173.x Copyright © 2001 International Society of Nephrology Terms and Conditions

Figure 1 Renal function parameters of rats with different cold ischemia times 24 hours after transplantation. (A) Serum creatinine concentration. (B) Serum urea concentration. Both parameters expressed in mg/dL as means ± SEM. Kidney International 2001 60, 1173-1181DOI: (10.1046/j.1523-1755.2001.0600031173.x) Copyright © 2001 International Society of Nephrology Terms and Conditions

Figure 2 Acute tubular necrosis (ATN) score in renal grafts exposed to different cold ischemia times 24 hours after transplantation. All results are expressed as means ± SEM. The incremental increases between 2 and 12 hours are significant at every step. Kidney International 2001 60, 1173-1181DOI: (10.1046/j.1523-1755.2001.0600031173.x) Copyright © 2001 International Society of Nephrology Terms and Conditions

Figure 3 Effect of cold preservation times and subsequent reperfusion injury on amount of different leukocyte populations and corresponding leukocyte integrins. Data are expressed as a number of positively labeled cells counted in 15 randomly chosen fields of view in sections of grafts exposed to different cold ischemia times. (A) Number of infiltrating neutrophils (▪; granulocytes). (B) Number of LFA-1+ cells (▪). (C) Number of ED-1+ monocytes/macrophages (▪). (D) Number of VLA-4+ cells (▪). All results are expressed as means ± SEM. *P ≤ 0.001. NK, native kidney. Kidney International 2001 60, 1173-1181DOI: (10.1046/j.1523-1755.2001.0600031173.x) Copyright © 2001 International Society of Nephrology Terms and Conditions

Figure 4 Effect of cold preservation times and subsequent reperfusion injury on amount of recipient immunocompetent antigen-presenting cells (APCs) that infiltrate the graft. (A) Number of MHC class II-positive cells (▪). (B) Number of OX-62 dendritic cells (▪). (C) Number of ED-2+ macrophages with dendritic phenotype (▪). All results are expressed as means ± SEM. NK, native kidney. Kidney International 2001 60, 1173-1181DOI: (10.1046/j.1523-1755.2001.0600031173.x) Copyright © 2001 International Society of Nephrology Terms and Conditions

Figure 5 Expression and distribution of PECAM-1 24 hours after transplantation in: isografts with (A) 2 hours, (B) 6 hours, (C) 12 hours, and (D) 24 hours of cold ischemia. Original magnification ×200. Kidney International 2001 60, 1173-1181DOI: (10.1046/j.1523-1755.2001.0600031173.x) Copyright © 2001 International Society of Nephrology Terms and Conditions

Figure 6 Immunostaining for VCAM-1 24 hours after transplantation in renal isografts exposed to (A) 2 hours and (B) 6 hours cold preservation. Original magnification ×400. Kidney International 2001 60, 1173-1181DOI: (10.1046/j.1523-1755.2001.0600031173.x) Copyright © 2001 International Society of Nephrology Terms and Conditions

Figure 7 Immunostaining for tissue factor 24 hours after transplantation in renal isografts. Vascular expression in grafts exposed to (A) two hours and (C) six hours cold preservation. Intraglomerular and peritubular expression in grafts exposed to (B) two hours and (D) six hours cold preservation. Original magnification ×400. Kidney International 2001 60, 1173-1181DOI: (10.1046/j.1523-1755.2001.0600031173.x) Copyright © 2001 International Society of Nephrology Terms and Conditions