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Identification and characterization of two genes (MIP-1 beta, VE-CADHERIN) implicated in acute rejection in human heart transplantation : Use of murine models in tandem with cDNA arrays
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Edité par CCSD ; American Heart Association -
International audience. 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-1 beta (MIP-1 beta), 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-1 beta and downregulation of VE-cadherin may strongly participate in human acute heart rejection.
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