腺相关病毒介导PSA基因转染DC的实验研究PPT.ppt

腺相关病毒介导PSA基因 转染DC的实验研究;Adaptive and Innate Immune Systems ;Ag /MHC Class II;Dendritic cells express a variety of cell surface molecules which play key roles in their interactions with T cells. The MHC molecules present antigenic fragments (peptides) to T cells in a form that can be recognized in a highly specific fashion by the appropriate T cell receptor, which is unique for every T cell and every antigen. The costimulatory molecules (CD80 and CD86) provide a second-signal that amplifies the antigen recognition process. Accessory molecules like ICAMs and LFAs enhance the physical interaction between T cells and DCs. ;前列腺癌细胞大量表达PSA;Gene delivery loading;Vectors for Gene delivery Retroviruses (Retrov) (8kb, RNA, enveloped) Adenoviruses (Ad) (35kb, DNA, non- enveloped) Adeno-Associated Virus (AAV) (5kb, DNA, non- enveloped) ;Retroviruses (Retrov) (8kb, RNA, enveloped) 1) Transgene inactivation by DNA methylation． 2) Causes cancer: proviral chromosomal integration activates proto-oncogene ． 3) Retrovirus vectors are easily destroyed, easily inactivated. ;　Adenoviruses (Ad) (35kb, DNA, 　　　　　　　non-enveloped) -are highly immunogenic, causing inflammation and cell death，that is the non-therapeutic immune response -can not infect non-dividing cells（Ｇ０） ;Adeno-Associated Virus (AAV) (5kb, DNA, non-enveloped) non-pathogenic low immune response chromosomal integration long term expression in vivo infect dividing and non-dividing cells ;;Ag /MHC Class II; Protocol of Dendritic Cells Generation;rAAV infection ;day 3 day5 day 7;;;;;;;;51Cr释放法检测CTL对靶细胞杀伤的抗原特异性;51Cr释放法检测CTL对靶细胞杀伤的MHC I 类限制性 ;研究结论 成功构建了AAV/PSA质粒，其能够高效转染DC； AAV/PSA基因成功整合入DC染色体内； DC内成功表达PSA蛋白；; AAV/PSA转染DC后，仅1周即可诱导得到大量、高活性的CTL； 所获得CTL对PSA阳性的前列腺癌细胞具有特异、高效的细胞毒杀伤效应。 ;Thank you!