1、腺相關病毒介導PSA基因轉染DC的實驗研究尤長宣 羅榮城 梁衛(wèi)江南方醫(yī)科大學南方醫(yī)院腫瘤中心The Oncology Department of Nanfang Hospital, the Southern Medical University, Guangzhou Adaptive and Innate Immune Systems Skin & Mucous Membranesrapidly regenerating surfaces, peristaltic movement, mucociliary escalator, vomiting, flow of urine/tears, c

2、oughingCellular and Humoral Defenseslysozyme, sebaceous/mucous secretions, stomach acid, commensal organisms, complement proteins, phagocytosis, natural killer cellsInvasion& infectionBarriersInnate immunityAdaptive immunityInflammationCellular and Humoral DefensesAntibodies, cytokines, helper T cel

3、ls, cytotoxic T cells（CTL）MK-0069-01Ag /MHC Class IIAg/ MHC Class ICD4ActivationCytokinesCD8TARGET LYSIS Activation（CTL）Dendritic cell- MHC Class I restricted- antigen specificAntigen presentationTumor antigenAntigen positive tumor cellDendritic cells express a variety of cell surface molecules whic

4、h 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 (

5、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. Dendritic Cell Properties 前列腺癌細胞大量表達PSAPSA是前列腺細胞內的一種特異性蛋白；前列腺癌標志物選擇PSA作為前列腺癌目標抗原Loading dendritic cells with the an

6、tigenProtein/peptide loadingGene delivery loading Disadvantages of Protein/peptide loadingT1/2 of proteins/peptides is very short. All are rapidly lost.2) Most protocols for generating cytotoxic T lymphocytes（CTL） take 3-5 weeks.Retroviruses (Retrov) (8kb, RNA, enveloped) 1) Transgene inactivation b

7、y DNA methylation2) 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 resp

8、onse-can not infect non-dividing cells（）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 rAAV infection Chromium release assayprimingday 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15GM-CSFIL-4IL-2 and IL-7Mixed with T-cells or PBLTNF-Structure of experimentsday 3day5day 7CTL Day 551Cr釋放法檢測CTL對靶細胞殺傷的抗原特異性51Cr釋放法檢測CTL對靶細胞殺傷的MHC I 類限制性 研究結論成功構