A study of molecular diagnosis and treatment for chronic cardiac allograft rejection.

本研究分为三个部分，描述如下。1 .我们利用实时定量RT-PCR技术测量了65种不同免疫基因在发生同种异体心脏移植血管病变（CAV）的再移植大鼠心脏中的表达动态。该再移植模型为研究慢性排斥反应的分子机制提供了更好的机会，因为所鉴定基因的表达模式与慢性排斥反应的发生之间的相关性比其他动物模型更为严格。综上所述，除了激活T细胞产生的细胞因子（IFN-γ和Fas配体）、血管平滑肌细胞生长因子（PDGF-β、TGF-β）在cav进展中的作用外，还指出了选择性趋化因子/受体系统（包括IP10-CXCR3、RANTES-CCR5和MCP1-CCR2）的重要性。将本发明的实时RT-PCR技术应用于重症肌无力的临床情况研究，为胸腺瘤细胞MHCⅱ类抗原表达异常抑制ⅱ类反激活因子基因在重症肌无力发病机制中的作用提供了更多的数据。这一结果也表明了该方法在临床诊断包括cav在内的多种疾病的可行性。3：采用β -肾上腺素能受体基因和肝细胞生长因子基因进行实验性基因治疗。在这两种情况下，通过将任何一种基因引入体内的整个心脏，缺血再灌注损伤都减轻了。由于CAV是一种全身性免疫抑制的局部疾病，因此可以想象，该技术将为临床治疗CAV提供途径。临床心脏移植在日本刚刚起步，本研究开发的技术不可能按原计划应用于慢性排斥反应的临床诊断。尽管如此，本研究为临床CAV的诊断和治疗提供了一些新的方法。少

This study was performed as three parts described as below.1 : We have measured the dynamics of 65 different immune gene expressions using real-time quantitative RT-PCR in retransplanted rat hearts developing cardiac allograft vasculopathy (CAV). This retransplantation model provided better opportunities to investigate the molecular mechanism, because the correlation between the occurrence of chronic rejection and the expression pattern of the identified gene was more stringent than those in other animal models. In summary of this part of the study, the importance of selective chemokine/receptor systems including IP10-CXCR3, RANTES-CCR5, and MCP1-CCR2 was indicated in addition to the implication of cytokines produced by activated T cells (IFN-γ and Fas ligand), growth factors for vascular smooth muscle cells (PDGF-β, TGF-β) in the progression of CAV.2 : The developed technology using real-time RT-PCR was applied to the investigation of Myasthenia Gravis in clinical situation and provid … More ed the data suggesting the implication of suppressed class II transactivator gene-expression of dysregulation of MHC class II antigen on thymoma cells in pathogenesis of the disease. This results also indicated the feasibility of this method in clinical diagnosis of various diseases includeing CAV.3 : Experimental gene therapy was performed using beta-adrenergic receptor gene and hepatocyte growth factor gene. In both cases, ischemia-reperfusion injury was attenuated by the introduction of either gene into a whole heart in vivo. It is conceivable that the technique should provide the way to treat CAV in clinical situation, because CAV is a local disorder under systemic immunosuppression.Clinical heart transplantation in Japan has just began and it was impossible to apply the technique developed in this study on the clinical diagnosis of chronic rejection as originally planed. Nonetheless, this study provided several new methods applicable to the diagnosis and the treatment of CAV in clinical situation. Less