1 / 23

HBsAg 阳性肝细胞的膜表面 HBsAg 抗原的检测

Parvogen. International. HBsAg 阳性肝细胞的膜表面 HBsAg 抗原的检测. Hep3B. HBsAg 阳性肝细胞. 91.3%. 93.5%. HepG2. 2.3%. Parvogen. International. Map of AAV Genome. HBsAb Fd. Cap. Parvogen. International. pHelper 质粒的改建. E4. Rep. E2A. 启动子. pHelper. VAI. Ori.

quinn-robinson
Download Presentation

HBsAg 阳性肝细胞的膜表面 HBsAg 抗原的检测

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Parvogen International HBsAg阳性肝细胞的膜表面HBsAg抗原的检测 Hep3B HBsAg阳性肝细胞 91.3% 93.5% HepG2 2.3%

  2. Parvogen International Map of AAV Genome

  3. HBsAb Fd Cap Parvogen International pHelper质粒的改建 E4 Rep E2A 启动子 pHelper VAI Ori

  4. A series of human cells infected by tropic AAV/AFPp-GFP virus （24 hours) Parvogen International HBsAg阳性 HBsAg阴性 Hep3B 肝细胞肝癌细胞 HepG2 肝细胞 HBsAg阴性 PBMC Hela LNcap Hs578T H2125 Lovo

  5. A series of human cells infected by tropic AAV/ALBp-GFP virus （24 hours) Parvogen International HBsAg阳性 HBsAg阴性 Hep3B 肝细胞肝癌细胞 HepG2 肝细胞 HBsAg阴性 PBMC Hela LNcap Hs578T H2125 Lovo

  6. Parvogen International Tropic AAV/ALBp-HLA-A2 virus infection （24 hours) HBsAg阳性肝细胞 HBsAg阴性肝细胞 未感染 感染 感染 5.6% 91.1% 1.4%

  7. Parvogen International Tropic AAV/AFPp-HLA-A2 virus infection （24 hours) HBsAg阳性Hep3B细胞 HBsAg阴性HepG2细胞 未感染 感染 感染 87.8% 4.7% 10.7%

  8. Uninfected cells AAV/HLA-2-infected cells uninfected cells AAV/HLA-2-infected cells Uninfected cells AAV/HLA-2-infected cells Parvogen International HLA-A2-expressed Primary Hepatic Carcinoma Cells Killed by the rAAV/AFP-infected-DC Pulsed CTL AAV/AFPp-HLA-A2- infected cells uninfected cells 90.1% 0.01% 90.1% 0.01% % killing 50 40 30 20 10 0

  9. Section II Parvogen International AAV-Rep78 function: interaction with HBV or AFP • Inhibit oncogene expression • Inhibit cervical cancer • Inhibit Human papillomavirus p97 promoter • Inhibit E6 & E7 oncogene • Enhance recombinant AAV production Can Rep78 inhibit HBV or AFP expression?

  10. Parvogen International Expression of HBsAg inhibited by tropic AAV/AFPp-Rep78 virus infection

  11. Parvogen International 68.9% Expression of HBeAg inhibited by tropic AAV/AFPp-Rep78 virus infection in HBV-positive PHC 感染前 24小时 48小时 72小时 96小时 84.6% 53.9% 10.7% 32.9%

  12. Parvogen International Expression of AFP inhibited by tropic AAV/ALBp-Rep78 virus infection

  13. Parvogen International Elevation of albumin by tropic AAV/ALBp-Rep78 virus infection in Hep3B cells 未感染前 48小时 72小时 96小时 96.6 50.8% 45.6% 22.3% 12.6%

  14. Parvogen International AAV Rep78与HBV基因的相互作用具有剂量依赖效应 Enh I X基因启动子 Enh II 前C/C启动子 Rep78 (μg):0 0.2 0.5 0 0.2 0.5 0 0.2 0.5 0 0.2 0.5

  15. Parvogen International Inhibition of HBV-C and –X Transcription by Rep78 C+Rep78 C X+Rep78 X+GST X 1.0ug 1.0ug

  16. 甲胎蛋白启动子 Parvogen International Section III Functions of expressed interferon in rAAV-transfeted hepatic cells

  17. ALBp ALBp AFPp IFN-γ基因 IFN-γ基因 IFN-α基因 Poly A Poly A Poly A TR TR TR TR TR TR rAAV/Albp-IFN-γ rAAV/Albp-IFN-γ rAAV/Albp-IFN-α Parvogen International 构建携带干扰素基因的嗜肝性重组腺相关病毒

  18. Parvogen International 52.9% 77.2% 52.9 25.7% 12.1% rAAV在肝细胞中表达IFN-α抑制HBsAg和HBeAg的表达 rAAV在肝细胞中表达IFN-a对HBeAg表达的影响 0小时 48小时 72小时 96小时

  19. Parvogen International rAAV在感染的黑猩猩中表达人IFN-a AAV/Albp-IFN-α Virus, 1011 copies Control AAV

  20. Parvogen International Expressed IFN-γ promote expression of MHC class I on rAAV-infected Hep3B cells IFN-γ MHC class I \ \ % 2.5% 感染前 感染AAV/AFPp -IFN-γ病毒后 52.3% 33.7% 12 hr 78.0% 48.5% 48 hr 85.2% 76.0% 96 hr

  21. Parvogen International rAAV在肝细胞中表达的IFN-γ增强抗原特异性，MHC class I限制性CTL的杀伤活性 70 60 50 40 30 20 10 0 CTL杀伤率 A B C D E F G H HBsAg+肝细胞：B: HepG2 C: +anti-HLA I D:+INF-r Hep3B: F: HBsAg+肝细胞 G:+anti-HLA I H:+INF-r

  22. Parvogen International Conclusions • Generate rAAV with liver cell-specific promoters. • Gernerate liver-tropic rAAV. • Recover MHC class I expression by transfection of liver-tropic rAAV. • AAV Rep78 may become new anti-HBV infection or hepatic cancer drug. • Expressed INF-αcan inhibit HBV expression in the rAAV-transfected liver cells. • The liver-tropic AAV carrying IFN-Αcan express in animal models. • Expressed IFN-γcan promote expression of the HLA class I antigen in the rAAV-transfected liver cells.

  23. Thank You

More Related