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Novel Chemotactic-Antigen DNA Vaccine against Cancer 新型肿瘤趋化抗原基因疫苗

Novel Chemotactic-Antigen DNA Vaccine against Cancer 新型肿瘤趋化抗原基因疫苗. Shuren Zhang 张叔人. Cancer Institute, Hospital Chinese Academy of Medical Sciences Peking Union Medical Collage. Regulation of Cytokines secretion by secondary lymphoid tissue chemokine (SLC).

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Novel Chemotactic-Antigen DNA Vaccine against Cancer 新型肿瘤趋化抗原基因疫苗

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  1. Novel Chemotactic-Antigen DNA VaccineagainstCancer 新型肿瘤趋化抗原基因疫苗 Shuren Zhang 张叔人 Cancer Institute, Hospital Chinese Academy of Medical Sciences Peking Union Medical Collage

  2. Regulation of Cytokines secretion by secondary lymphoid tissue chemokine (SLC) Intratumoral SLC administration promotes Th1 cytokine and antiangiogenic chemokine release and a decline in immunosuppressive mediators Th1 cytokine : IFN-g, GM-CSF, IL-12, MIG, and IP-10 Immunosuppressive mediators: PGE2, IL-10, and TGF-b angiogenic factor: VEGF The Journal of Immunology, 2000, 164: 4558–4563. Steven M. Dubinett

  3. Ag Her2,TERT PSA,PAP CEA,HPV SLC CMV Fc Enzyme Cut Sites Chemotactic-Antigen DNA Vaccine (CADV) 趋化抗原基因疫苗

  4. B DC Th MHC-II CCR7 FcR CTL MHC-I Tumor T CCR7 CMV SLC Ag Fc Effect Mechanisms of CADV Gene transfer and expression Recruiting DC and T cell ; catching Ag by ligands; Ag presentation T and B cell activation, anti-Tumor Effects Injection or Gene gun EP Antigen presentation by DCs through ligand-mediated internalization can be enhanced up to 10,000-fold over fluid-phase antigen pinocytosis

  5. Mouse Model of CADV HPV16E7 Her2/P53 CMV mSLC Fc

  6. Mouse Model of CADV (pSLC-E7-Fc) E7

  7. RT-PCR and Western Blotting Analysis (A) RT-PCR detection of B16F10 cells transfected with various recombinant plasmids. DNA marker (M),Lane 1 (vector control), Lane 2 (SLC fragment), Lane 3 (Sig-E7 fusion fragment), Lane 4 (SLC-E7 fusion fragment),Lane 5 (Sig-Fc fusion fragment), Lane 6 (SLC-Fc fusion fragment), Lane 7 (Sig-E7-Fc fusion fragment), Lane 8 (SLC-E7-Fc fusion fragment). (B) Western blotting analysis of B16F10 transfectants. SLC-E7-Fc fusion protein (50kD), E7-Fc fusion protein (38kD),SLC-E7 fusion protein (22kD).

  8. Chemotaxis Assay by Chemotaxis Chamber pSLC-E7-Fc pcDNA

  9. Group Tumor free pC pSLC-Fc psig-E7 pSLC-E7 psig-E7-Fc pSLC-E7-Fc 0/8 (0%) 0/8 (0%) 4/8 (50%) 3/8 (38%) 3/8 (38%) 8/8 (100%) Therapeutic immunization with pSLC-E7-Fc induces C3 tumor regression HPV16-E7 as tumor antigen. E7+C3 tumor cell subcutaneous inoculated in C57BL/6 mice on day0. The tumor bearing mouse immunized by gene gun with different gene vaccine on day 6, day 13 and day 20.

  10. Therapeutic immunization with pSLC-E7-Fc induces TC-1 tumorregression HPV16-E7 as tumor antigen. E7+TC-1 tumor cell subcutaneous inoculated in C57BL/6 mice on day0. The tumor bearing mouse immunized by gene gun with different gene vaccine on day 15, day 22 and day 28.

  11. Tumor incidence after TC-1 tumor recharlenged Recharlenge with 1.8 x 105 TC-1, after 90days later of first TC-1 inoculation.

  12. PSA-mPAP-PSM (3P) Her2/P53 CMV SLC Fc Enzyme Cut Sites Prostate Cancer Chemotactic-Antigen DNA Vaccine 前列腺癌趋化抗原基因疫苗

  13. human SLC with Chemotactic Activity for mouse immune cells Human PBMC Mouse spleen cell A and B: Human PBMC; C and D: mouse spleen cell; A and C: Negative control (200×); B and D: SLC-3P-Fc (200×)

  14. Transfection with pSLC-3P-Fc chemoattracts lymphocytesto the immunization site A B C pathology H&E staining of dermis tissues sections from C57BL/6 mice, non-treated mice (A), treated with pC (B) or pSLC-3P-Fc (C). Images are representative of multiple microscopic fields observed in at least three mice per group.

  15. Immunotherapy effects of human pSLC-3P-Fc gene vaccine on mouse model B16-3P tumor cell subcutaneous inoculated in C57BL/6 mice on day0. The tumor bearing mouse immunized by gene gun with different gene vaccine on day 4, day 9 and day 14. 3P: PSA-PAP-PSM; Target: B16-3P;

  16. PBMC HLA-A0201 CADV transfected MLC Restimulate Induction of human CTL against cancer by CADV in vitro Day 0 Day 1 Day 6 Day 13 MLC: Mixed Lymphocyte Culture

  17. SLC-3P-Fc DNA Vaccine induced CTL by MLC Effector:Target=60:1 A0201 + - + + + + + + hPSA - + - - + - - + hPSM - + - - - + - + hPAP - + - - - - + + A: Stimulating cell tansfected by pSLC-3P-Fc B: Stimulating cell tansfected by pcDNA

  18. hTERT PSA,PAP,PSM CMV SLC Fc Enzyme Cut Sites Universal Chemo-Antigen DNA Vaccine (CADV) 广谱趋化抗原基因疫苗

  19. SLC-TERT-Fc DNA Vaccine induced human CTL by MLC The CTLs induced by pSLC-T-Fc-transfected PBMC, pC-transfected PBMC, and non-transfected PBMC were tested for cytolytic activity against MCF-7, SK-BR3, U2OS/T, U2OS, LNCap, PC-3M, SK-OV-3, and T2 .

  20. Assessment of involvement of NK cells, CD4+ and CD8+T cells in antitumor effect CD8 CD4 NK FIGURE 8 – Assessment of involvement of NK cells, CD4+ and CD8+T cells in antitumor effect. Animals were s.c. challenged with 5 x 104 B16/Te cells on day 0, then treated with B16/SLC-Te-Fc or B16/C plus anti-4-1BB (n = 4) as Figure 6a described. Cell subsets were depleted by the injection of GK1.5 (for CD4+ T cells), 2.43 (for CD8+ T cells) ascites fluids (100 ml/mouse) on days 2, 4 and 6, then twice a week before killing. For NK cells depletion, asialo-GM1 antibodies (200 mg/mouse) were injected for 5 consecutive days beginning on day 2, then twice a week before killing. The control Ig was injected in the same way. (a) Abrogation of CTL-mediated cytotoxicity. (b) The s.c. tumor size was measured twice per week.

  21. Evaluation of therapeutic effect on established pulmonary metastases by CADV-tumor + anti-41BB B16/SLC-Te-Fc + anti-4-1BB B16/SLC-Te-Fc + Ig B16/C + anti-4-1BB B16/C +Ig C57BL/6 mice were challenged via tail vein with 1 x 105 B16/Te tumor cells on day 0 (n = 5), and and were treated with CADV modified tumor cells immunization on days 7 and 11 followed by either rat Ig or anti-4-1BB MAbs administration (i.p.) on days 8, 11 and 14. The lung metastatic nodules were counted on day 23. *p < 0.001 versus B16/C + Ig and B16/C + anti-4-1BB by Student paired t-test; **p < 0.05 versus B16/CCL21-Te-Fc 1 Ig by Student paired t-test.

  22. The Advantageous Features of Chemotactic-Antigen DNA Vaccine 1. The CADV could facilitate the co-localization of DC, T, B and NK cells by SLC and induce effective cell-mediated immune response and humoral immune response . 2. CADV fusion protein can be forcefully captured by DC (Fc/FcR). This approach can efficiently increase antigen presentation by DCs, to induce both antigen-specific Th and CTL. 3. Tumor antigen specific memory T cells can be highly induced by CADV to prevent tumor recurrence and metastasis. 4. CADV is a recombinant plasmid DNA. It is suitable for industrialisation. 5. CADV is a vaccine platform. Tumor or non-tumor vaccines can be easily made by this system.

  23. Publications 1、Specific antitumor immune response induced by a novel DNA vaccine composed of multiple CTL and T helper cell epitopes of prostate cancer associated antigen， Immunology Letters 99:85-93,2005 2、Enhancement of Antitumor Immunity by a Novel Chemotactic Antigen DNA Vaccine Encoding Chemokine and ultiepitope of Prostate Tumor Associate Antigens, Immunology， 117: 419-430, 2006 3、Enhancement of DNA vaccine potency by sandwiching antigen-coding gene between secondary lymphoid tissue chemokine (SLC) and IgG Fc fragment genes, Cancer Biology and Therapy，Apr 20;5(4), 2006 4、Enhanced anti-tumor effect against human telomerase reverse transcriptase (hTERT) by vaccination with chemotactic-hTERT gene-modified tumor cell and its combination with anti-4-1BB monoclonal antibodies, International Journal of Cancer119, 2006 5、Induction of protective and therapeutic antitumour: immunity using a novel tumour-associated antigen-specific DNA vaccine,Immunology and Cell Biology84, 1–8 , 2006 6、Potent Systemic Antitumor Immunity Induced by Vaccination with Chemotactic-Prostate Tumor Associated Antigen Gene-Modified Tumor Cell and Blockade of B7-H1，Journal of Clinical Immunology, 27(1):117-130, 2007 7、Synergistic antitumor effect of chemotactic-prostate tumor associated antigen gene-modified tumor cell vaccine and anti-CTLA-4 mAb in murine tumor model，Immunology Letters, 113:90–98, 2007 8、A novel chemotactic-antigen DNA vaccine against cancer, Future Oncology, Apr;4(2):299-303,2008

  24. Thank you ! Lin Chen Sun Wenxin Qin Hanjun Lin Xiaoyan Liu Rongzhi

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