以重组腺相关病毒为基础的肝脏疾病的基因治.pptVIP

* * * * * * * * * 以重组腺相关病毒为基础的肝脏疾病的基因治疗 张大志1, 2，周军1,3，尤红1,4 ，周卫平1, 2，夏建川1,3 ，Paul L. Hermonat1, Kahm Maha11, Jay L. Mehta11, 刘勇1 1. Gene Therapy Center; University of Arkansas for Medical Sciences. 72205, USA 2. 重庆医科大学附属第二医院感染科, 中国 3.中山大学肿瘤医院生物治疗中心, 中国 4.首都医科大学北京友谊医院肝病研究中心, 中国 AAV Gene Therapy Infect dividing and non-dividing cells. Adeno-Associated Virus (AAV) A helper dependent. Non-enveloped. Parvovirus, linear 4.7 kb, ss DNA. Non-pathogenic. Chromosomal integration. Long term expression in vivo. Low immune response. Recovery: rAAV-Pulsed Dendritic Cells to Elicit CTL Killing Activity against Hepatic Diseases Section I TARGET LYSIS - MHC Class I Restricted - Antigen Specific AAV-DC治疗原理 Ag /MHC Class II Ag/ MHC Class I Activation Activation CD4 Target cells: tumor cells CD8 Dendritic Cells (DC) rAAV infection 52% 48% 48% 43% K-cells A Liver B C Breast D Lung AAV/PSA-pulsing of DC results in a high killing of primary prostate cancer cells (A, B, C & D patients) The PSA-specific CTL Activity in Animal Model One Lung Cancer (adeocarcinoma) Case Treated with the CTL Stimulated by AAV/CEA and AAV/Her2 Virus-infected DC for 2 years This patient has survived for almost 5 years. Failure of CTL Killing Activity against primary hepatic carcinoma cells 0 10 20 30 40 50 60 70 80 90 100 AAV/α-FP target - Anti- MHC K562 HCC Cells (HBV+ , α-FP+) Why ? AAV/HBV-S Low Expression of MHC Class I (HLA I)Molecules in Primary Hepatic Carcinoma Cells Chronic active hepatitis with piecemeal necrosis and parenchymal inflammation Histologic features of HCV Cirrhosis Hepatocellular carcinoma 激活的抗原特异性，MHC Class I限制性CTL攻击靶细胞机制 AAV/GFP infection rAAV-GFP in mouse Lymphocyte (60-80%) not granular Not monocyte 利用AAV载体恢复肝细胞的HLA I类抗原表达 重组腺相关病毒载体 TR TR p5 ,CMVp启动子 HLA I-A2或报道 基因(GFP, Luci.) Poly A 白蛋白 启动子 甲胎蛋白启动子 ALBp: 白蛋白启动子 AFPp: 甲胎蛋白启动子 Generation of Recom