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(Circulation. 2005;111:2636-2644.)
© 2005 American Heart Association, Inc.

Transplantation

Identification and Characterization of Two Genes (MIP-1ß, VE-CADHERIN) Implicated in Acute Rejection in Human Heart Transplantation

Use of Murine Models in Tandem With cDNA Arrays

A.L.S. Roussoulières, MD, PhD; O. Raisky, MD; L. Chalabreysse, MD; G. Dureau, MD; C. Cerutti, PhD; C. Thieblemont, MD, PhD; P. Boissonnat, MD; L. Sebbag, MD, PhD; J.-F. Obadia, MD, PhD; J. Ninet, MD; O. Bastien, MD, PhD; F. Thivolet-Bejui, MD, PhD; J.L. McGregor, PhD, DrBH

From INSERM U331/EA 1582 Faculté de Médecine RTH Laennec, Lyon, France (A.L.S.R., C.C., J.L.M.); Departments of Cardiac Transplantation (A.L.S.R., P.B., L.S., O.B.), Cardiac Surgery (O.R., G.D., J.-F.O., J.N.), and Pathology (L.C., F.T.-B), Hôpital Cardiologique Louis Pradel, Lyon, France; Department of Hematology, Hôpital Lyon Sud, Lyon, France (C.T.); EMIU 0226, Lyon, France (L.S.); and Cardiovascular Division, King’s College, and Thrombosis Research Institute, Genomics and Atherothrombosis, London, UK (J.L.M.).

Correspondence to Ana L.S. Roussoulières, MD, PhD, Hôpital Cardiologique Louis Pradel, Department of Cardiac Transplantation, 28 Avenue du Doyen Lépine, 69003 Lyon, France. E-mail ana.roussoulieres{at}chu-lyon.fr

Received June 8, 2003; de novo received December 10, 2003; revision received January 8, 2005; accepted January 19, 2005.

Background— Genes and mechanisms of action involved in human acute rejection after allogeneic heart transplantation remain to be elucidated. The use of a murine allograft model in tandem with cDNA arrays and quantitative real-time polymerase chain reaction (Q-PCR) can greatly help in identifying key genes implicated in human heart acute rejection.

Methods and Results— Hearts from Balb/c mice were either not transplanted or transplanted heterotopically in the abdomen of Balb/c (isografts) and C57BL/6 (allografts) mice. Histological analysis showed acute rejection only in allografts. Total RNA was extracted from isografts (n=3), allografts (n=4), and not transplanted hearts (n=4); reverse transcribed; and labeled with P32. Each probe was hybridized to cDNA macroarrays. Eight genes were overexpressed and 7 genes were underexpressed in allografts compared with isografts. Macrophage inflammatory protein-1ß (MIP-1ß), an overexpressed gene, and VE-cadherin, an underexpressed gene, were validated by immunohistochemistry and Q-PCR in the murine models. Genes of interest, validated in the 3 murine groups, were then investigated in human heart tissues. Immunohistochemistry and Q-PCR performed on endomyocardial biopsies after heart transplantation showing no rejection (n=10) or grade IB (n=10) or IIIA (n=10) rejection, according to International Society of Heart and Lung Transplantation criteria, confirmed the results obtained from the murine model.

Conclusions— We have demonstrated that the upregulation of MIP-1ß and downregulation of VE-cadherin may strongly participate in human acute heart rejection.

Key Words: genes • immunohistochemistry • polymerase chain reactions • rejection • transplantation