title: Identification and characterization of two genes (MIP-1beta, VE-CADHERIN) implicated in acute rejection in human heart transplantation: use of murine models in tandem with cDNA arrays.
abstract: 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.
  
  
  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-1beta (MIP-1beta), 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-1beta and downregulation of VE-cadherin may strongly participate in human acute heart rejection.
authors.affiliation: INSERM U331/EA 1582 Faculté de Médecine RTH Laennec, Lyon, France. ana.roussoulieres@chu-lyon.fr
authors.forename: A L S
authors.initials: AL
authors.lastname: Roussoulières
authors.forename: O
authors.initials: O
authors.lastname: Raisky
authors.forename: L
authors.initials: L
authors.lastname: Chalabreysse
authors.forename: G
authors.initials: G
authors.lastname: Dureau
authors.forename: C
authors.initials: C
authors.lastname: Cerutti
authors.forename: C
authors.initials: C
authors.lastname: Thieblemont
authors.forename: P
authors.initials: P
authors.lastname: Boissonnat
authors.forename: L
authors.initials: L
authors.lastname: Sebbag
authors.forename: J-F
authors.initials: JF
authors.lastname: Obadia
authors.forename: J
authors.initials: J
authors.lastname: Ninet
authors.forename: O
authors.initials: O
authors.lastname: Bastien
authors.forename: F
authors.initials: F
authors.lastname: Thivolet-Bejui
authors.forename: J L
authors.initials: JL
authors.lastname: McGregor
mesh_terms: D000818:Animals; D015703:Antigens, CD; D015820:Cadherins / Q000032:analysis / Q000235:genetics*; D054407:Chemokine CCL4; D020869:Gene Expression Profiling; D006084:Graft Rejection / Q000235:genetics*; D016027:Heart Transplantation / Q000009:adverse effects*; D006801:Humans; D007150:Immunohistochemistry; D019402:Macrophage Inflammatory Proteins / Q000032:analysis / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D023421:Models, Animal; D020411:Oligonucleotide Array Sequence Analysis; D014184:Transplantation, Homologous; D014185:Transplantation, Isogeneic; D015854:Up-Regulation; D000099152:Cadherin 5
chemical_list: D015703:Antigens, CD; D015820:Cadherins; D054407:Chemokine CCL4; D019402:Macrophage Inflammatory Proteins; D000099152:Cadherin 5