In Press, 1995, Transplantation Proceedings copyright by Appleton and Lange.

Molecular Activation Markers in Rejection Diagnosis

von Willebrand E, Häyry P, Taskinen E.

Transplantation Laboratory and Fourth Department of Surgery, University of Helsinki, Finland

 

Reprint requests to:	Dr. E. von Willebrand
			Transplantation Laboratory
			University of Helsinki
			PO Box 21 (Haartmaninkztu 3)
			SF 00014 Helsinki, Finland

Supported by the Finnish Academy of Science.

 

INTRODUCTION

The characteristic early findings in acute cellular rejection of human kidney allografts are generation of inflammation, appearance of activated lymphoid cells and, later activation of monocytes into macrophages. Concomitantly activation markers appear on the lymphoid cells, expression of intercellular adhesion molecules on different graft components is increased and Class II antigens are upregulated also on graft parenchymal cells. The appearance of IL-2 receptor expressing cells and an increase in Class II positive lymphocytes in the graft have been the most common measurable markers of lymphoid activation. The induction of these activation markers is well established in acute cellular rejection [1-3].

Normal kidneys and stable grafts express varying amounts of adhesion molecules on endothelial cells of capillaries and larger vessels [4-6].The relationship between ICAM-1 and Class II induction in acute rejection of renal transplants has been demonstrated previously [7, 8], but the relationship between other adhesion molecule and activation marker induction has not been studied. The expression of cellular adhesion molecules; intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), endothelial leucocyte adhesion molecule-1 (ELAM-1), platelet endothelial cell adhesion molecule (PECAM-1) is regulated by several cytokines, which are produced early during lymphoid and monocytic activation [1-3]

We have studied the induction of lymphoid activation markers CD25 (IL2 receptors), CD69 (AIM, early lymphoid activation), HLA ClassII molecules and the monocytic marker CD68 and their relationship to the expression of cellular adhesion molecules; intercellular adhesion molecule-1(ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), platelet-endothelial cell adhesion molecule-1 (PECAM-1) and endothelial leucocyte adhesion molecule-1 (ELAM-1), in renal transplants with acute rejection and compared the findings to transplants with no rejection.

 

MATERIALS and METHODS

The expression of activation markers CD25 (IL2 receptors) on lymphoid cells, CD69 (AIM, early lymphoid activation), HLA ClassII molecules and the monocytic marker CD68, and the expression of adhesion molecules ICAM-1, VCAM-1, PECAM-1 and ELAM-1, was evaluated in 15 renal transplants with acute rejection and in 10 transplants with no rejection. The number of CD2 positive cells was also analyzed. Diagnosis of rejection was based on clinical findings, aspiration cytology and on histological biopsies. All rejections were early, occurring during the first month posttransplant. Evaluation of activation markers and adhesion molecules was done from frozen sections of histological biopsies using indirect immunoperoxidase staining with monoclonal antibodies. The positively stained cells were calculated in all different components of the graft per high power visual field. In this study the reactions of the interstitial cells, this including both infiltrating interstitial inflammatory cells and capillary endothelial cells, have been analyzed.

 

RESULTS AND DISCUSSION

[ Table 1.] demonstrates the expression of activation markers and adhesion molecules during rejection and no rejection. In the 15 grafts with acute rejection there was interstitial inflammation, with increased CD2 lymphocytes, 40 ±23, and induction of activation markers, IL2R positive cells 12 ±5, CD69 lymphocytes 5 ±4, CD68 monocytes 70 ±8 and ClassII positive interstitial cells, lymphocytes and monocytes, 60 ±41. There was also strong induction of all adhesion molecules on the interstitial cells; ICAM-1, VCAM-1 and PECAM-1 were 97 ±20, 80 ±38, 69 ±16 respectively and ELAM-1 was induced to 12 ±9.

In the 10 grafts with no rejection, there were no interstitial infiltrates, CD2 lymphocytes low 3 ±3, and no induction of lymphoid activation markers, no IL2-receptors and no CD69 lymphocytes. Also monocytes and ClassII positive cells were low, 4 ±3 and 9 ±9, respectively per high power visual field. ICAM-1, VCAM-1 and PECAM-1 expressing cells were low: 13 ±10, 16 ±13, 16 ±13. ELAM-1 expression was very low, 1 ±1. The differences between the two groups were all statistically significant, p. <0.005.

The major target of acute rejection in kidney allografts, is the vascular endothelium, especially of the intertubular capillaries. The expression of vascular adhesion molecules on the cell membrane are involved with the early phase of lymphoid activation and serve as adhesion receptors for lymphocytes and other inflammatory cells [1]. Cellular adhesion molecules and their ligands also have an important role in leucocyte extravasation and in the development of cellular infiltrates in different tissues, also in transplants. This study demonstrates, that in acute rejection molecular activation markers are induced on infiltrating inflammatory cells and are related to a strong induction of adhesion molecules on different components of the kidney graft, and that these phenomena can be used in rejection diagnosis.

 

REFERENCES

  1. Hall BM, Duggin GG, Philips J, Bishop GA, Horvarth JS, Tiller DJ. Lancet 1984; II: 247.

     

  2. Hancock WW, Gee D, De Moerloose P, et.al.. Transplantation 1985; 39: 430.

     

  3. von Willebrand E, Häyry P. Transplant Proc 1987; 19:1644.

     

  4. Springer TA. Nature 346;425,1990.

     

  5. Brockmeyer C, Ulbrecht M, Schendel DJ, et al. Transplantation 55;610,1993.

     

  6. Faull RJ, Russ GR. Transplantation 48:226,1989.

     

  7. von Willebrand E, Loginov R, Salmela K, et al. Transplant Proc 25:870,1993.

     

  8. von Willebrand E, Krogerus L, Salmela K, et al. Transplant Proc 27;917,1995.

     


 

TABLES

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Table 1: Activation markers and adhesion molecules in rejection diagnosis.

 
  CD2 CD25 CD69 CD68 ClassII IC ICAM VCAM PECAM ELAM
NOR 3 ±3 0 ±0 0 ±0 4 ±3 9 ±9 13 ±10 16 ±13 16 ±13 1 ±1
REJ 40 ±23* 12 ±5* 5 ±4* 70 ±8* 60 ±41* 97 ±20* 80 ±38* 69 ±16* 12 ±9*
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* p<0.005

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Last Modified: April 03, 1996 11:35:02 AM