胃癌特异性纳米疫苗的制备及其生物学活性的研究
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摘要
胃癌是严重威胁我国人民健康的恶性肿瘤,目前对晚期胃癌的治疗手段主要有手术切除和化疗,但均不能有效解决肿瘤转移,复发等问题。免疫治疗是肿瘤治疗的新方法。随着现代细胞和分子生物学的发展,已经基本明确机体对肿瘤的免疫识别和免疫调节的过程。在此基础上,研究特异性肿瘤疫苗正逐渐成为肿瘤生物治疗的重要手段。但由于缺乏特异抗原和有效的抗原递呈,以及MHC的限制和肿瘤异质性等原因,使现有肿瘤疫苗的使用范围和效果相当有限。MG7—Ag是本研究所在八十年代发现的特异性较高的胃癌标志物,抗原分析表明其抗原决定簇位于糖链上。由于糖抗原免疫原性弱,纯化、制备困难,且不易诱导T细胞免疫反应,用分子模拟多肽替代糖抗原进行免疫治疗是解决问题的方法之一,并在动物模型中取得很好的效果。本研究所用噬菌体随机肽库筛选得到了MG7—Ag的模拟表位,抗体竞争结合抑制实验证实该模拟表位可
Background Gastric cancer is a common malignancy in manycountries in the world, especially in China. Because of lowefficiency of the conventional treatment, the prognosis ofpatients with gastric cancer is still poor. Cancer vaccine is aneffective method for cancer therapy that was demonstrated by alot of preclinical and clinical research. However, thedevelopment of gastric cancer vaccine was hampered by the lackof specific tumor antigen. MG7-Ag, discovered by our institute,is a new marker of gastric cancer. The epitope of MG7~Ag was foundto locate in a sugar chain, which suggested that the MG7-Ag wasa kind of carbohydrate antigen. The mimicry peptide of MG7-Ag wasobtained previously by screening the phage display librarytechnology in our lab. In vivo experiment showed that the mimicrypeptide could mimic the primary antigen effectively, whichindicated that mimicry peptide of MG7-Ag was a suitable candidatemolecule for the development of gastric cancer vaccine.
CpG oligodeoxynucleotides (CpG ODN) is a kind of new high efficient immune adjuvant. Recent research has shown: CpG DNA and/or synthetic CpG oligodeoxynucleotides (CpG ODN) having potent adjuvant activity for a wide range of antigens, It can induce both strong innate and specific immunity and shows promising adjuvant effect for tumor antigen. Besides, CpG ODN can be used alone to activate the innate immunity and trigger a tumor-specific immune response. Of particular interest is their improved activity when closely associated with the antigen.Most of antigen mimitopes are composed of 8-12 peptides, which are degraded easily in vivo and require vector to protect them, so as to enhance the ability of antigen presentation and activate specific immune response. Nanoemulsion is a hypoviscosity, single optically isotropic and thermodynamically stable liquid solution with smaller diameter of 0.1-100nm,, Composed of biological materials, it also has reduced toxicity and is more efficiency. It is potentially becoming a focus point as a delivery formulation for peptides and DNA drug. Nanoemulsion has many advantages as following: (1)It' s thermodynamically stable and can be sterilized by fileration, it also is easilly to be prepared and stored. (2)It can increase the dissolving capacity of different drugs which are difficult to be dissolved in water or in lipid solutions. It is hypoviscosity so there is no pain when be used as injection solution. ?It' s good dispersity make it easy to be absorbed by human body and so improve
the drug' s efficiency. (4)It can protect the drugs which are easilly hydrolysised, and so prolong the function time of the drug. (5)It has target characteristics and can deliver the drug to defferent organs through controlling the diameter of the nanoemulsion. (6)It can schlep ectogenic peptides into cytoplasm through endocytosis and break through the MHC limit through cross-presentation.Another approach to improve the immunogenicity of the peptides vaccine is through combination of multiple epitopes. Multi-epitope vaccines induces specific immune response aganist each epitope in the vaccine. And the potency of repeated-epitope vaccine could be improved because the epitope' s copy is increased and Thl response is enhanced. When several epitopes are combined together, flanking sequence will affect the potency of the vaccine. Therefore, it is crucial to insert spacers between epitopes. Proper spacer can prvent mutual interference between epitopes and unspecific binding with MHC molecules. What more, it can also improve the synergy funcion between epitopes, enhance the binding of epitopes and MHC molecules and partially prevent the hydrolysis of protease to peptides.The purpose of this study is to construct a nanovaccine co-encapsulated with a gastric cancer specific antigen MG7-Ag mimotope peptides and adjuvant CpG ODN 1645 using new nanotechnology as nanoemulsion and evaluate its immunocompetence.
Nanovaccine was prepared using magnetic ultrasound methods. BALB/c mice were immunized and the in vivo effectiveness was evaluated using tumor challenge assay. Part I : Study on preparation and characteristic of nanoemulsion. Objective To study the way of preparation of nanoemulsion with proper diameter and stable charateristics. Methods Plot phase diagram was used to study the composing of nanoemulsion and nanoemulsion was prepared using magnetic and ultrasonic technique. Its chacteristic was tested by centrifuging, dye and electron microscope, etc. MTT assay was used to assay its cytotoxicity. Results We successefully prepared the nanoemulsion according the phase diagram. Dye assay showed that it was water in oil (W/0) with average diameter of 20~30nm, high stability and low cytotoxicity. Conclusion The emulsion we prepared fit the standard of nanoemulsion. It was stable and has ideal diameter. Part II: Preparation of gastric cancer specific nanovaccine and study of its immunocompetence Objective To develop a nanovaccine coencapsulated with one MG7~Ag mimotope and CpG adjuvent using new nano-technology and valuate its immune efficacy using animal experiment. Methods Encapsulate synthesized MG7-Ag mimotope peptide and CpG ODN into nanoemulsion using magnetic and ultrasonic technique. HPLC was used to determine encapsulation efficiency. BALB/c mice were immunized intravenously with nanovaccine. Serum titer of MG7 antibody was determined by ELISA assay. MTT, ELISPOT was performed to test the cytotoxicity of
spleen cells. The protective and tumor inhibiting effect of the nanovaccine was evaluated by tumor cell challenge assay. Results MG7 — Ag nanovaccine was successfully constructed with high stability and encapsulation efficiency. It could induce both cellular and humour immune response to MG7-Ag in mice. Tumor cell challenge showed that the tumor masses formed in the mice immunized with coencapsulation nanovaccine became markedly smaller than those formed in the mice of control groups and the tumor inhibition rate of co-encapsulation group is obiviously high than other groups. Conclusion Nanovaccine based on the coencapsulation of one MG7-Ag mimotope and CpG is immunogenic which can induce specific immunity response against tumor in mice. And the vaccine is partially protective. Partlll Preparation of gastric cancer specific nanovaccine and study of its immunocompetence Objective To develop a nanovaccine coencapsulated with multiple MG7-Ag mimotopes and CpG adjuvent using new nano-technology and compare its immune efficacy with one mimotope nanovaccine using animal experiment. Methods Encapsulate synthesized multiple MG7 mimotope peptide and CpG ODN into nanoemulsion using magnetic and ultrasonic technique. Dialyse method was used to sterilize, HPLC to determine encapsulation efficiency. BALB/c mice were immunized with multi-epitope nanovaccine. ELISA assay was used to determine serum titer of MG7 antibody. MTT, ELISPOT was performed to test the cytotoxicity of spleen cells. The protective and tumor
Background Gastric cancer is a common malignancy in manycountries in the world, especially in China. Because of lowefficiency of the conventional treatment, the prognosis ofpatients with gastric cancer is still poor. Cancer vaccine is aneffective method for cancer therapy that was demonstrated by alot of preclinical and clinical research. However, thedevelopment of gastric cancer vaccine was hampered by the lackof specific tumor antigen. MG7-Ag, discovered by our institute,is a new marker of gastric cancer. The epitope of MG7~Ag was foundto locate in a sugar chain, which suggested that the MG7-Ag wasa kind of carbohydrate antigen. The mimicry peptide of MG7-Ag wasobtained previously by screening the phage display librarytechnology in our lab. In vivo experiment showed that the mimicrypeptide could mimic the primary antigen effectively, whichindicated that mimicry peptide of MG7-Ag was a suitable candidatemolecule for the development of gastric cancer vaccine.
CpG oligodeoxynucleotides (CpG ODN) is a kind of new high efficient immune adjuvant. Recent research has shown: CpG DNA and/or synthetic CpG oligodeoxynucleotides (CpG ODN) having potent adjuvant activity for a wide range of antigens, It can induce both strong innate and specific immunity and shows promising adjuvant effect for tumor antigen. Besides, CpG ODN can be used alone to activate the innate immunity and trigger a tumor-specific immune response. Of particular interest is their improved activity when closely associated with the antigen.Most of antigen mimitopes are composed of 8-12 peptides, which are degraded easily in vivo and require vector to protect them, so as to enhance the ability of antigen presentation and activate specific immune response. Nanoemulsion is a hypoviscosity, single optically isotropic and thermodynamically stable liquid solution with smaller diameter of 0.1-100nm,, Composed of biological materials, it also has reduced toxicity and is more efficiency. It is potentially becoming a focus point as a delivery formulation for peptides and DNA drug. Nanoemulsion has many advantages as following: (1)It' s thermodynamically stable and can be sterilized by fileration, it also is easilly to be prepared and stored. (2)It can increase the dissolving capacity of different drugs which are difficult to be dissolved in water or in lipid solutions. It is hypoviscosity so there is no pain when be used as injection solution. ?It' s good dispersity make it easy to be absorbed by human body and so improve
the drug' s efficiency. (4)It can protect the drugs which are easilly hydrolysised, and so prolong the function time of the drug. (5)It has target characteristics and can deliver the drug to defferent organs through controlling the diameter of the nanoemulsion. (6)It can schlep ectogenic peptides into cytoplasm through endocytosis and break through the MHC limit through cross-presentation.Another approach to improve the immunogenicity of the peptides vaccine is through combination of multiple epitopes. Multi-epitope vaccines induces specific immune response aganist each epitope in the vaccine. And the potency of repeated-epitope vaccine could be improved because the epitope' s copy is increased and Thl response is enhanced. When several epitopes are combined together, flanking sequence will affect the potency of the vaccine. Therefore, it is crucial to insert spacers between epitopes. Proper spacer can prvent mutual interference between epitopes and unspecific binding with MHC molecules. What more, it can also improve the synergy funcion between epitopes, enhance the binding of epitopes and MHC molecules and partially prevent the hydrolysis of protease to peptides.The purpose of this study is to construct a nanovaccine co-encapsulated with a gastric cancer specific antigen MG7-Ag mimotope peptides and adjuvant CpG ODN 1645 using new nanotechnology as nanoemulsion and evaluate its immunocompetence.
Nanovaccine was prepared using magnetic ultrasound methods. BALB/c mice were immunized and the in vivo effectiveness was evaluated using tumor challenge assay. Part I : Study on preparation and characteristic of nanoemulsion. Objective To study the way of preparation of nanoemulsion with proper diameter and stable charateristics. Methods Plot phase diagram was used to study the composing of nanoemulsion and nanoemulsion was prepared using magnetic and ultrasonic technique. Its chacteristic was tested by centrifuging, dye and electron microscope, etc. MTT assay was used to assay its cytotoxicity. Results We successefully prepared the nanoemulsion according the phase diagram. Dye assay showed that it was water in oil (W/0) with average diameter of 20~30nm, high stability and low cytotoxicity. Conclusion The emulsion we prepared fit the standard of nanoemulsion. It was stable and has ideal diameter. Part II: Preparation of gastric cancer specific nanovaccine and study of its immunocompetence Objective To develop a nanovaccine coencapsulated with one MG7~Ag mimotope and CpG adjuvent using new nano-technology and valuate its immune efficacy using animal experiment. Methods Encapsulate synthesized MG7-Ag mimotope peptide and CpG ODN into nanoemulsion using magnetic and ultrasonic technique. HPLC was used to determine encapsulation efficiency. BALB/c mice were immunized intravenously with nanovaccine. Serum titer of MG7 antibody was determined by ELISA assay. MTT, ELISPOT was performed to test the cytotoxicity of
spleen cells. The protective and tumor inhibiting effect of the nanovaccine was evaluated by tumor cell challenge assay. Results MG7 — Ag nanovaccine was successfully constructed with high stability and encapsulation efficiency. It could induce both cellular and humour immune response to MG7-Ag in mice. Tumor cell challenge showed that the tumor masses formed in the mice immunized with coencapsulation nanovaccine became markedly smaller than those formed in the mice of control groups and the tumor inhibition rate of co-encapsulation group is obiviously high than other groups. Conclusion Nanovaccine based on the coencapsulation of one MG7-Ag mimotope and CpG is immunogenic which can induce specific immunity response against tumor in mice. And the vaccine is partially protective. Partlll Preparation of gastric cancer specific nanovaccine and study of its immunocompetence Objective To develop a nanovaccine coencapsulated with multiple MG7-Ag mimotopes and CpG adjuvent using new nano-technology and compare its immune efficacy with one mimotope nanovaccine using animal experiment. Methods Encapsulate synthesized multiple MG7 mimotope peptide and CpG ODN into nanoemulsion using magnetic and ultrasonic technique. Dialyse method was used to sterilize, HPLC to determine encapsulation efficiency. BALB/c mice were immunized with multi-epitope nanovaccine. ELISA assay was used to determine serum titer of MG7 antibody. MTT, ELISPOT was performed to test the cytotoxicity of spleen cells. The protective and tumor
引文
1. Hewitt HB, Black ER, Walder AS. A critique of the evidence for active host defence against cancer, based on personal studies of 27 rnurine tumours of spontaneous origin. Br J Cancer. 1976; 33(3): 241-259.
2. Burnett FM. Immunological aspects of malignant disease. Lancet. 1976; 1(7501): 1171-1174.
3. Willemsen R, Ronteltap C, Heuveling M, et al. Redirecting human CD4+ T lymphocytes to the MHC class I-restricted melanoma antigen MAGE-Al by TCR alphabeta gene transfer requires CDSalpha. Gene Ther. 2005; 12(2): 140-6.
4. Pedersen AE, Thorn M, Gad M, et al. Phenotypic and functional characterization of clinical grade dendritic cells generated from patients with advanced breast cancer for therapeutic vaccination. Scand J immunol. 2005; 61(2): 147-56.
5. Dillman RO. Monoclonal antibodies in the treatment of malignancy: basic concepts and recent developments. Cancer Invest. 2001; 19(8): 833-841
6. Coiffier B. Rituximab in combination with CHOP improves survival in elderly patients with aggressive non-Hodgkin's lymphoma. Semin Oncol. 2002; 29(2 suppl 6): 18-22.
7. Ligibel JA, Winer EP. Trastuzumab/chemotherapy combination in metastatic breast cancer. Semin Oncol. 2002; 29(3 suppl 11): 38-42.
8. Gruaz-Guyon A, Raguin O, Barber J et al. Recent advances in pretargeted radioimmtmotherapy. Curt Med Chem. 2005; 12(3): 319-38.
9. Francis RJ, Sharma SK, Springer C, et al. A phase I trial of antibody directed enzyme prodrug therapy (ADEPT) in patients with advanced colorectal carcinoma or other CEA producing tumours. Br J Cancer. 2002; 87(6): 600-7.
10. Tatalick LM, Gerard CJ, Takeya R, Price DN, Thorne BA, Wyatt LM, Anklesaria P. Safety characterization of HeLa-based cell substrates used in the manufacture of a recombinant adeno-associated virus-HIV vaccine. Vaccine. 2005; 23(20): 2628-38.
11. Simone Mocellin, Carlo Riccardo Rossi, Donato Nitti. Cancer vaccine development: on the way to break immune tolerance to malignant cells Experimental Cell Research. 2004; 29 (9) 267-278
12. Qian FIN, Liu GZ, Cao SJ, et al. The experimental study of ovarian carcinoma vaccine modified by human B7-1 and IFN-gamma genes. Int J Gynecol Cancer. 2002; 12(1): 80-5.
13. Svane IM, Pedersen AE, Johnsen HE, et al. Vaccination with p53-peptide-pulsed dendritic cells, of patients with advanced breast cancer: report from a phase I study. Cancer Immunol Imnunother. 2004; 53(7): 633-41.
14. Welters MJ, Filippov DV, van den Eeden SJ, et al. Chemically synthesized protein as turnour-specific vaccine: immunogenicity and efficacy of synthetic HPV16 E7 in the TC-1 mouse tumour model. Vaccine. 2004; 23(3): 305-11.
15. Dillman RO. Monoclonal antibodies in the treatment of malignancy: basic concepts and recent developments. Cancer Invest. 2001; 19(8): 833-841
16. Oka. Y, Tsuboi. A, Elissccva. O. A., et al. WT1 as a novel target for cancer immunotherapy. Curr Cancer Drug Targets. 2002; 2(1): 45-54.
17. Schreiber H. Tumour Immunology. In: Paul WE ed. Fundamental Immunology. 4th ed. Paul Lippincott-Ravcn Publishers, Philadelphia: 1999. 1237-1270.
18. Rosenbcrg SA. Cancer vaccines base, don the identification of genes encoding cancer regression antigens. Immunol Today. 1997; 18: 175-182.
19. Theobald M, Biggs J, Dittmcr D, et al. Targeting p53 as a general tumour antigen. Proc Natl Acad Sci USA. 199: 5; 92:11993-11997.
20. Hakomori. S., Zhang, Y. Glycosphingolipid antigens and cancer therapy. Chem Bio. 1997; 4: 97-104.
21. Gorelik E., Galili U., Raz A. On the role of cell surface carbohydrates and their binding protein (lectins) in tumour metastasis. Cancer Metastasis Rev. 2001; 20: 245-277.
22. Appelmelk BJ, Simoons SI, Negrini R, et al. Potentil role of molecular mimicry between helicobacter pylorilipopolysaccharide and host lewis blood group antigens in autoimmunity. Infect Immun. 1996; 64: 2031-2040.
23. Burchell JM, Mungul A, Taylor-Papadimitriou J. O-linked glycosylation in the mammary gland: changes that occour during malignancy. J Mammary Gland Biol. Neoplasia. 2001; 6: 355-364.
24. Hralzumi S, Takasaki S, Ochuchi N, et al. Altered glycosylation of membrane glycoproteins associated with human mammary carcinoma. J. Cancer Res. 1992; 83: 1063-1072.
25. Goldblatt D. Conjugate vaccines [editorial]. Clin Exp Immunol. 2000; 119: 1-3.
26. Ben-Yedidia T, Beignon AS, Partidos CD, et al. A retro-inverso peptide analogue of influenza virus hemagglutinin B cell epitope 91-108 induces a strong mucosal and systemic immune response and confers protection in mice after intranasal immunization. Mol Immunol. 2002; 39: 323-331.
27. Apostolopoulos V, Sandrin MS, Mckenzie IF. A carbohydrate peptide minics: effect on MUCI cancer immunotherapy. J Mol Med. 1999; 77: 427-436.
28. Alfonso M, Diaz A, Hernandez AM, et al. An anti-idintype vaccine elicits a specific response to N-glycolylsialic acid residues of glycoconjugates in melanoma patients. J Immunol. 2002; 168: 2523-2529.
29. Haurum JS, Aresquell G, Lellouch AC, et al. Recognition of carbohydrate major histocompatibility complex class I-restricted glycopeptidespecific cytotoxic T lymphocytes. J Exp Med. 1994; 180: 739-744.
30. Jenen T, Hansen P, Galli SL, et al. Carvohydrate and peptide specificity of MHC class Ⅱ-restricted T cell hybridomas raised against an O-glycosylated self peptide. J Immunol. 1997; 158: 3769-3778.
31. Jenen T, Galli SL, Mouritsen S, et al. T cell recognition of Tn-glycosylated peptide antigens. Eur J Immunol. 1996; 26: 1342-1349.
32. Ludewig B, Ochsenbein AF, Odermatt B, et al. Immunotherapy with dendritic cells directed against tumor antigens shared with normal host cells results in severe autoimmune disease. J Exp Med 2000; 191:795-803.
33. Disis ML, Rinn K, Knutson KL, et al. Flt3 ligand as a vaccine adjuvant in association with HER - 2/ neu peptide - based vaccines in patients with HER - 2/ neu - overexpressing cancers [J]. Blood,2002, 99 (8) : 2845-2850.
34. Hara S, Nagai H, Miyake H, et al. Secreted type of modified interleukin-18 gene transduced into mouse renal cell carcinoma cells induces systemic tumor immunity. J Urol. 2001; 165(6 Pt 1): 2039-43.
35. Qian HN, Liu GZ, Cao SJ, et al. The experimental study of ovarian carcinoma vaccine modified by human B7-1 and IFN-gamma genes. Int J Gynecol Cancer. 2002; 12(l):80-5.
36. Ge NL, Ye SL, Zheng N. Prevention of hepatocellular carcinoma in mice by IL-2 and B7-1 genes co-transfected liver cancer cell vaccines. World J Gastroenterol. 2003; 9(10):2182-5.
37. Willimsky G, Blankenstein T. Interleukin - 7/ B7.1 - encoding adenoviruses induce rejection of transplanted but not nontransplanted tumors. Cancer Res, 2000,60:685.
38. Antonia SJ, Seigne J, Diaz J, et al. Phase I trial of a B7 - 1 (CD80) gene modified autologous tumor cell vaccine in combination with systemic interleukin - 2 in patients with metastatic renal cell carcinoma. J Urol 2002,167 (5): 1995-2000.
39. Guckel B, Stumm S, Rentzsch C. A CD80-transfected human breast cancer cell variant induces HER-2/neu-specific T cells in HLA-A*02-matched situations in vitro as well as in vivo. Cancer Immunol Immunother. 2005; 54(2): 129-40.
40. Kato K, Cantwell MJ, Sharma S, et al. Gene transfer of CD40 - ligand induces autologous immune recognition of chronic lymphocytic leukemia B cells. J Clin Invest, 1998, 5 :1133-1141.
41. Ito D, Ogasawara K, Iwabuchi K, et al. Induction of CTL responses by stimultaneous administration of liposomal peptide vaccine with anti-CD40 and anti-CTLA4 tnAb.J Immunol.2000; 164(3): 1230-1235.
42. Krieg A. CpG oligonucleotides as immune adjuvants. Ernst Schering Res Found Workshop.2000; 105-118
43. Bukowski R, Ernstoff MS, Gore ME, et al. Pegylated interferon alfa-2b treatment for patients with solid tumours: a phase I / II study.J Clin Oncol.2002; 20(18): 3841-3849.
44. Nagliera E, Lopea M, Lelli G, et al. Interleukin-2 interferon-alpha and medroxyprogesterone acetate in metastatic renal cell carcinoma Anticancer Res.2002; 22(5), 3045-3051.
45. Rini BI, Weiberg V, Bok R, et al. Prostate-specific antigen kinetics as a measure of the biologic effect of granulocytemacrophage colonstimulating factor in patients with serologic progression of prostate cancer.J Clin Oncol.2003;21(l): 99-105.
46. Slingluff CL Jr, Petroni GR, Yamshchikov GV. Immunologic and clinical outcomes of vaccination with a multiepitope melanoma peptide vaccine plus low-dose interleukin-2 administered either concurrently or on a delayed schedule. J Clin Oncol. 2004 Nov 15; 22(22): 4474-85.
47. Dakappagari NK, Lute KD, Rawale S, et al. Conformational HER-2/neu B-cell epitope peptide vaccine designed to incorporate two native disulfide bonds enhances tumor cell binding and antitumor activities. Biol Chem. 2005 Jan 7; 280(1): 54-63. Epub 2004 Oct 26.
48. Ayyoub M, Mazarguil H, Monsarrat B, et al. A structure - based aproach to designing non - natural peptides that can activate antimelanoma cytotoxic T cells. J Biol Chem, 1999,274:10227.
49. Waeckerle-Men Y, Gander B, Groettrup M. Delivery of tumor antigens to dendritic cells using biodegradable microspheres. Methods Mol Med. 2005; 109:35-46.
50. You Z, Hester J. Rollins L, et al. A Retrogen Strategy for Presentation of an Intracellular Tumor Antigen as an Exogenous Antigen by Dendritic Cells Induces Potent Antitumor T helper and CTL Responses. Cancer Research, 2001,61:197.
51. Tan M C, Mommaas A M, Drijfhout J W, et al. Mannose receptor mediated uptake of antigens strongly enhanced HLA class II restricted antigen presentation by cultured dendritic cells. Eur. J. Immunol, 1999,23:3217.
52. Strome SE, Voss S. Wilcox R, et al. Strategies for antigen loading of dendritic cells to enhance the antitumor immune response. Cancer Res, 2002,62 (6): 1884 ~ 1889.
53. Vieweg J, Jackson A. Modulation of antitumor responses by dendritic cells. Springer Semin Immunopathol. 2005; 26(3): 329-41.
54. Nikitina EY, Chada S, Muro - Cacho C, et al. An effective immunization and cancer treatment with activated dendritic cells transduced with full - length wild - type p53. Gene The, 2002 , 9 (5): 345-352.
55. Trakatelli M, Toungouz M, Lambermont M, et al. Immune characterization of clinical grade-dendritic cells generated from cancer patients and genetically modified by an ALVAC vector carrying MAGE minigenes. Cancer Gene Then 2005; 21:4893-7.
56. Kawada M, Ikeda H, Takahashi T, et al. Vaccination of fusion cells of rat dendritic and carcinoma cells prevents tumor growth in vivo. Int J Cancer. 2003; 105(4): 520-6.
57. Huttner KG, Breuer SK, Paul P, et al. Generation of potent anti-tumor immunity in mice by interleukin-12-secreting dendritic cells. Cancer Immunol Immunother. 2005 Jan; 54(1): 67-77.
58. Lespagnard L, Mettens P, De Smedt T, et al. The immune response induced in vivo by dendritic cells is dependent on B7-1 or B7-2, but the inhibition of both signals does not lead to tolerance. Int Immunol. 1998; 10(3):295-304.
59. Munn DH, Sharma MD, Mellor AL. Ligation of B7-1/B7-2 by human CD4+ T cells triggers indoleamine 2,3-dioxygenase activities in dendritic cells. J Immunol. 2004; 172(7): 4100-10.
60. Zitvogel L, Regnault A, Lozier A, et al. Eradication of established murine tumors using a novel cell-free vaccine: dendritic cell-derived exosomes. Nat Med. 1998; 4(5): 594-600.
61. Amigorena S. Cancer immunotherapy using dendritic cell-derived exosomes. Medicina (B Aires). 2000; 60 Suppl 2:51-4.
62. Morse MA, Garst J, Osada T, et al. A phase I study of dexosome immunotherapy in patients with advanced non-small cell lung cancer. Transl Med. 2005; 3(1):9.
63. Park JH, Kim CJ, Lee JH, et al. Effective immunotherapy of cancer by DNA vaccination. Mol Cells, 1999,9 (4): 384.26
64. De Giovanni C, Nanni P, Forni G. The prospects for cancer gene therapy. Int J Immunopharmacol, 2000,22 (12): 1025.
65. Sumida SM, McKay PF, Truitt DM. Recruitment and expansion of dendritic cells in vivo potentiate the immunogenicity of plasmid DNA vaccines. J Clin Invest. 2004 Nov; 114(9): 1334-42.
66. Zhang S, Li WF, Zhang HJ, et al. Antitumor reactivity of splenocytes primed in vivo with dendritic-cell-based vaccine and secondarily activated with a cocktail of cytokines in vitro. Exp Oncol. 2004; 26(3): 243-5.
67. Coelho M, Gauthier P, Pugniere M, et al. Isolation and characterisation of a human anti-idiotypic scFv used as a surrogate tumour antigen to elicit an anti-HER-2/neu humoral response in mice. Br J Cancer. 2004; 90(10): 2032-41.
68. Malavaud B, Pedron S, Sordello S, et al. Direct FGF receptor 1 activation through an anti-idiotypic strategy mimicks the biological activity of FGF-2 and inhibits the progression of the bladder carcinoma derived from NBT-II cells. Oncogene. 2004; 23(40): 6769-78.
69. Avin E, Haimovich J, Hollander N. Anti-idiotype x anti-CD44 bispecific antibodies inhibit invasion of lymphoid organs by B cell lymphoma. J Immunol. 2004; 173(7): 4736-43.
70. Nicholson S, Bomphray CC, Thomas H, et al. A phase I trial of idiotypic vaccination with HMFG1 in ovarian cancer. Cancer Immunol Immunother. 2004; 53(9):809-16.
71. Binder RJ, Blachere NE, Srivastava PK. Heat shock protein - chaperoned peptides but not free peptides introduced into the cytosol are presented efficiently by major histocompatibility complex I molecules. J Biol Chem, 2001,276 (20) 17163~ 17171.
72. Meng SD, Gao T, Gao GF, Tien P. HBV-specific peptide associated with heat-shock protein gp96. Lancet. 2001; 357(9255): 528-9
73. Srivastava PK. Immunotherapy for human cancer using heat shock protein-Peptide complexes. Curr Oncol Rep. 2005; 7(2): 104-8.
74. Minev B, Hipp J, Firat H, S et al. Cytotoxic T cell immunity against telomerase reverse transcriptase in humans. Proc Natl Acad Sci USA. 2000; 97(9):4796-801.
75. Scappaticci FA, Nolan GP. Induction of anti-tumor immunity in mice using a syngeneic endothelial cell vaccine. Anticancer Res. 2003 Mar-Apr; 23(2B): 1165-72.
76. Susan S, Smyth, Cam Patterson Tiny dancers: the integrin-growth factor nexus in angiogenic signaling. J Cell Biology. 2002; 158(1): 17-21.
77. HAN Quan-Li, DING Jie, FAN Dai-Ming, et al. Selection of mimic epitope of gastric cancer associated antigen by using peptide library. J Fourth Mil Med Univ 2000,21(9) 1162-1164.
78. Liu DW, Tsao YP, Kung Jt, et al. Recombinant adeno-associated virus expressing human papillomavirus type 16 E7 peptide DNA fused with heat shock protein DNA as a potential vaccine for cervical. J Virol. 2000,74(6): 2888-2894.
79. Villen J, de Oliveira E, Nunez JI, et al. Towards a multi-site synthetic vaccine to foot-and-mouth disease: addition of discontinuous site peptide mimic increases the neutralization response in immunized animals Vaccine. 2004 9; 22(27-28): 3523-9.
80. Haicbeur N, Bismuth E, Bosset S, et al. The B subunit of Shiga toxin fused to a tumor antigen elicits CTL and targets dendritic cells to allow MHC class I-restricted presentation of peptides derived from exogenous antigens. J Immunol, 2000, 165(6): 3301-3308.
81. Kate K, Cantwell MJ, Sharma S, et al. Gene transfer of CD40-lig2 and induces autologous immune recognition of chronic lymphocytic leukemia B cells. J Clin Invest, 1998; 5: 1133～1141.
82. Hao X, Shao Y, Ren X, et al. Induction of specific CTL by MAGE-3/CEA peptide-pulsed dendritic cells from HLA-A2/A24(+) gastrointestinal cancer patients. J Cancer Res Clin Oncol. 2002; 128(9): 507-15.
83. Nori, K. D. et al. Tat-conjugated synthetic macromolecules facilitate cytoplasmic drug delivery to human ovarian carcinoma cells. Bioconugate. Chem. 2003, 14, 44-50.
84. Fonseca DP, Frofichs J, Singh M, et al. Induction of antibody and T-cell responses by immunization with ISCOMS containing the 38-kilodalton protein of Mycobacterium tuberculosis[J], Vaccine, 2000, 19(1): 122-131.
85. Brazolot Millan CL, Weeratna R, Krieg AM, et al. CpG DNA can induce strong Thl humeral and cell-mediated immune responses against hepatitis B surface antigen in young mice. Proc Nail Acad Sci USA 1998; 95: 15553-8.
86. Michael J. McCluskie, Risini D, et al. Parenteral and mucosal prime-boost immunization strategies in mice with hepatitis B surface antigen and CpG DNA. FEMS Immunology and Medical Microbiology. 2002; 32: 179-185.
87. Yamamoto S, Yamamoto T, Iho S, et al. Activation of NK cell [human and mouse] by immunostimulatory DNA sequence. Springer Semin Immunopathol. 2000; 22: 35-43.
88. C. Hirunpetcharat, J. Wipasa, S. Sakkhachomphop, et al. CpG oligodeoxynucleotide enhances immunity against blood-stage malaria infection in mice parenterally immunized with a yeast-expressed 19 kDa carboxyl-terminal fragment of Plasmodium yoelii merozoite surface protein-1 [MSP119] formulated in oil-based Muntanides. Vaccine. 2003; 21: 2923-2932.
89. Ningshu Liu, Noriko Ohnishi, Lin Ni, et al. CpG directly induces T-bet expression and inhibits IgG1 and IgE switching in B cells. Nature Immunology. 2003; 4: 687-693.
90. Gramzinski RA, Doolan DL, Sedagah M, et al. Interleukin-12- and gamma interferon-depondent protection against malaria conferred by CpG oligodeoxynucleotide in mice. Infect Immun. 2001; 69: 1643-9.
91. Sparwasser T, Koch ES, Vabulas RM, et al. Bacterial DNA and immunostimulatory CpG oligonucleotides trigger maturation and activation of murine dendritic cells. Eur J Immunol 1998; 28: 2045-54.
92. Wagner H, Hacker H, Lipford GB. Immunostimulatory DNA sequences help to eradicate intracellular pathogens. Springer Semin Immunopathol. 2000; 22: 147-52.
93. Tighe H, Takabayashi K, Schwartz D, et al. Conjugation of immunostimulatory DNA to the short ragweed allergen arab a 1 enhances its immunogenicity and reduces its allergenicity. J Allergy Clin Immunol. 2000; 106: 124-34.
94. Tighe H, Takabayashi K, Schwartz D, et al. Conjugation of protein to immunostimulatory DNA results in a rapid, long-lasting and potent induction of cell-mediated and humoral immunity. Eur J Immunol 2000; 30: 1939-47.
95. Morin C, Barratt G, Fessi H, et al. Improved intracellular deliver of a muramul dipeptide analog by means of nanocapsules. Int J Immunopharmacol. 1994; 16: 451-456.
96. Katare YK, Panda AK, Lalwani K, Haque IU, Ali MM. Potentiation of immune response from polymer-entrapped antigen: toward development of single dose tetanus toxoid vaccine. Drug Deliv. 2003; 10(4): 231-8.
97. Codard G, Boutorrine AS, Saison BE, et al. Antisense effects of cholesterol-oligodeoxynucleotide conjugates associated with poly (alkylcyanoacrylate) nanoparticles. Eur J Biochem. 1995; 232: 404-410.
98. Feng SS, Mu L, Win KY, et al. Nanoparticles of biodegradable polymers for clinical administration of paclitaxel. Curr Med Chem. 2004; 11(4): 413-24.
99. Murthy, N. et al. A macromolecular delivery vehicle for proteinbased vaccines: acid-degradable protein-loaded microgels. Proc. Natl. Acad. Sci. U. S. A. 2003 (100), 4995-5000
100. Desai MP, Labhasetwar V, Amidon GL, et al. Gastrointestinal uptake of biodegradable microparticles: effect of particle size. Pharm Res. 1996; 13(12): 1838-1845.
101. Liu Z, Xiao Y, Chen YH. Epitope-vaccine strategy against HIV-1: today and tomorrow. Immunobiology. 2003; 208(4): 423-8.
102. Markwin P, Velders. Defined flanking spacers and enhanced proteolysis is essential for eradication of established tumors by an epitopesstring DNA vaccine. J Immunol. 2001, 166: 5366-5373.
103. Desmezieres E, Jacob Y, Saron MF. Lyssavirus glycoproteins expressing immunologically potent B cell and. cytotoxic T lymphocyte epitopes as prototypes for multivalent vaccine. J Gene Virol. 1999; 80: 2343.
104. Mireille A, Jean-Pierre K. Recombinant fusion peptides containing single or multiple repeats of a ubiquitous T-helper epitope are highly immunogenic. Molecular Immunology. 1996; 33(13): 1017-1024.
105. Niedermann G, Geier E, Lucchiar/-Hartz M. The specificity of the proteasomes: impact on the MHC class Ⅰ processing and presentation of antigens. Immunol Rev. 1999; 172: 29.
106. Markwin P, Velders, et al. Defined flanking spacers andenhanced proteolysis is essential for eradication ofestablished tumors by an epitope string DNA vaccine. J Immunol. 2001; 166: 5366-5373.
107. Brain L, Claire C, Mark N, et al. A rational strategy to design multiepitope immunogens based on multiple Th lymphocyte epitopes. J Immunol. 2002; 168: 5499-5506.
108. Brain D, Livingston, Mark N, et al. Optomozation of epitope processing enhances immunogenicity of mulfiepitope DNA vaccines. Vaccine. 2001; 19: 4652-4660.
109. Lin Z, Wang Z, Chen YH. Predefined spacers between epitopes on a recombinant
2. Burnett FM. Immunological aspects of malignant disease. Lancet. 1976; 1(7501): 1171-1174.
3. Willemsen R, Ronteltap C, Heuveling M, et al. Redirecting human CD4+ T lymphocytes to the MHC class I-restricted melanoma antigen MAGE-Al by TCR alphabeta gene transfer requires CDSalpha. Gene Ther. 2005; 12(2): 140-6.
4. Pedersen AE, Thorn M, Gad M, et al. Phenotypic and functional characterization of clinical grade dendritic cells generated from patients with advanced breast cancer for therapeutic vaccination. Scand J immunol. 2005; 61(2): 147-56.
5. Dillman RO. Monoclonal antibodies in the treatment of malignancy: basic concepts and recent developments. Cancer Invest. 2001; 19(8): 833-841
6. Coiffier B. Rituximab in combination with CHOP improves survival in elderly patients with aggressive non-Hodgkin's lymphoma. Semin Oncol. 2002; 29(2 suppl 6): 18-22.
7. Ligibel JA, Winer EP. Trastuzumab/chemotherapy combination in metastatic breast cancer. Semin Oncol. 2002; 29(3 suppl 11): 38-42.
8. Gruaz-Guyon A, Raguin O, Barber J et al. Recent advances in pretargeted radioimmtmotherapy. Curt Med Chem. 2005; 12(3): 319-38.
9. Francis RJ, Sharma SK, Springer C, et al. A phase I trial of antibody directed enzyme prodrug therapy (ADEPT) in patients with advanced colorectal carcinoma or other CEA producing tumours. Br J Cancer. 2002; 87(6): 600-7.
10. Tatalick LM, Gerard CJ, Takeya R, Price DN, Thorne BA, Wyatt LM, Anklesaria P. Safety characterization of HeLa-based cell substrates used in the manufacture of a recombinant adeno-associated virus-HIV vaccine. Vaccine. 2005; 23(20): 2628-38.
11. Simone Mocellin, Carlo Riccardo Rossi, Donato Nitti. Cancer vaccine development: on the way to break immune tolerance to malignant cells Experimental Cell Research. 2004; 29 (9) 267-278
12. Qian FIN, Liu GZ, Cao SJ, et al. The experimental study of ovarian carcinoma vaccine modified by human B7-1 and IFN-gamma genes. Int J Gynecol Cancer. 2002; 12(1): 80-5.
13. Svane IM, Pedersen AE, Johnsen HE, et al. Vaccination with p53-peptide-pulsed dendritic cells, of patients with advanced breast cancer: report from a phase I study. Cancer Immunol Imnunother. 2004; 53(7): 633-41.
14. Welters MJ, Filippov DV, van den Eeden SJ, et al. Chemically synthesized protein as turnour-specific vaccine: immunogenicity and efficacy of synthetic HPV16 E7 in the TC-1 mouse tumour model. Vaccine. 2004; 23(3): 305-11.
15. Dillman RO. Monoclonal antibodies in the treatment of malignancy: basic concepts and recent developments. Cancer Invest. 2001; 19(8): 833-841
16. Oka. Y, Tsuboi. A, Elissccva. O. A., et al. WT1 as a novel target for cancer immunotherapy. Curr Cancer Drug Targets. 2002; 2(1): 45-54.
17. Schreiber H. Tumour Immunology. In: Paul WE ed. Fundamental Immunology. 4th ed. Paul Lippincott-Ravcn Publishers, Philadelphia: 1999. 1237-1270.
18. Rosenbcrg SA. Cancer vaccines base, don the identification of genes encoding cancer regression antigens. Immunol Today. 1997; 18: 175-182.
19. Theobald M, Biggs J, Dittmcr D, et al. Targeting p53 as a general tumour antigen. Proc Natl Acad Sci USA. 199: 5; 92:11993-11997.
20. Hakomori. S., Zhang, Y. Glycosphingolipid antigens and cancer therapy. Chem Bio. 1997; 4: 97-104.
21. Gorelik E., Galili U., Raz A. On the role of cell surface carbohydrates and their binding protein (lectins) in tumour metastasis. Cancer Metastasis Rev. 2001; 20: 245-277.
22. Appelmelk BJ, Simoons SI, Negrini R, et al. Potentil role of molecular mimicry between helicobacter pylorilipopolysaccharide and host lewis blood group antigens in autoimmunity. Infect Immun. 1996; 64: 2031-2040.
23. Burchell JM, Mungul A, Taylor-Papadimitriou J. O-linked glycosylation in the mammary gland: changes that occour during malignancy. J Mammary Gland Biol. Neoplasia. 2001; 6: 355-364.
24. Hralzumi S, Takasaki S, Ochuchi N, et al. Altered glycosylation of membrane glycoproteins associated with human mammary carcinoma. J. Cancer Res. 1992; 83: 1063-1072.
25. Goldblatt D. Conjugate vaccines [editorial]. Clin Exp Immunol. 2000; 119: 1-3.
26. Ben-Yedidia T, Beignon AS, Partidos CD, et al. A retro-inverso peptide analogue of influenza virus hemagglutinin B cell epitope 91-108 induces a strong mucosal and systemic immune response and confers protection in mice after intranasal immunization. Mol Immunol. 2002; 39: 323-331.
27. Apostolopoulos V, Sandrin MS, Mckenzie IF. A carbohydrate peptide minics: effect on MUCI cancer immunotherapy. J Mol Med. 1999; 77: 427-436.
28. Alfonso M, Diaz A, Hernandez AM, et al. An anti-idintype vaccine elicits a specific response to N-glycolylsialic acid residues of glycoconjugates in melanoma patients. J Immunol. 2002; 168: 2523-2529.
29. Haurum JS, Aresquell G, Lellouch AC, et al. Recognition of carbohydrate major histocompatibility complex class I-restricted glycopeptidespecific cytotoxic T lymphocytes. J Exp Med. 1994; 180: 739-744.
30. Jenen T, Hansen P, Galli SL, et al. Carvohydrate and peptide specificity of MHC class Ⅱ-restricted T cell hybridomas raised against an O-glycosylated self peptide. J Immunol. 1997; 158: 3769-3778.
31. Jenen T, Galli SL, Mouritsen S, et al. T cell recognition of Tn-glycosylated peptide antigens. Eur J Immunol. 1996; 26: 1342-1349.
32. Ludewig B, Ochsenbein AF, Odermatt B, et al. Immunotherapy with dendritic cells directed against tumor antigens shared with normal host cells results in severe autoimmune disease. J Exp Med 2000; 191:795-803.
33. Disis ML, Rinn K, Knutson KL, et al. Flt3 ligand as a vaccine adjuvant in association with HER - 2/ neu peptide - based vaccines in patients with HER - 2/ neu - overexpressing cancers [J]. Blood,2002, 99 (8) : 2845-2850.
34. Hara S, Nagai H, Miyake H, et al. Secreted type of modified interleukin-18 gene transduced into mouse renal cell carcinoma cells induces systemic tumor immunity. J Urol. 2001; 165(6 Pt 1): 2039-43.
35. Qian HN, Liu GZ, Cao SJ, et al. The experimental study of ovarian carcinoma vaccine modified by human B7-1 and IFN-gamma genes. Int J Gynecol Cancer. 2002; 12(l):80-5.
36. Ge NL, Ye SL, Zheng N. Prevention of hepatocellular carcinoma in mice by IL-2 and B7-1 genes co-transfected liver cancer cell vaccines. World J Gastroenterol. 2003; 9(10):2182-5.
37. Willimsky G, Blankenstein T. Interleukin - 7/ B7.1 - encoding adenoviruses induce rejection of transplanted but not nontransplanted tumors. Cancer Res, 2000,60:685.
38. Antonia SJ, Seigne J, Diaz J, et al. Phase I trial of a B7 - 1 (CD80) gene modified autologous tumor cell vaccine in combination with systemic interleukin - 2 in patients with metastatic renal cell carcinoma. J Urol 2002,167 (5): 1995-2000.
39. Guckel B, Stumm S, Rentzsch C. A CD80-transfected human breast cancer cell variant induces HER-2/neu-specific T cells in HLA-A*02-matched situations in vitro as well as in vivo. Cancer Immunol Immunother. 2005; 54(2): 129-40.
40. Kato K, Cantwell MJ, Sharma S, et al. Gene transfer of CD40 - ligand induces autologous immune recognition of chronic lymphocytic leukemia B cells. J Clin Invest, 1998, 5 :1133-1141.
41. Ito D, Ogasawara K, Iwabuchi K, et al. Induction of CTL responses by stimultaneous administration of liposomal peptide vaccine with anti-CD40 and anti-CTLA4 tnAb.J Immunol.2000; 164(3): 1230-1235.
42. Krieg A. CpG oligonucleotides as immune adjuvants. Ernst Schering Res Found Workshop.2000; 105-118
43. Bukowski R, Ernstoff MS, Gore ME, et al. Pegylated interferon alfa-2b treatment for patients with solid tumours: a phase I / II study.J Clin Oncol.2002; 20(18): 3841-3849.
44. Nagliera E, Lopea M, Lelli G, et al. Interleukin-2 interferon-alpha and medroxyprogesterone acetate in metastatic renal cell carcinoma Anticancer Res.2002; 22(5), 3045-3051.
45. Rini BI, Weiberg V, Bok R, et al. Prostate-specific antigen kinetics as a measure of the biologic effect of granulocytemacrophage colonstimulating factor in patients with serologic progression of prostate cancer.J Clin Oncol.2003;21(l): 99-105.
46. Slingluff CL Jr, Petroni GR, Yamshchikov GV. Immunologic and clinical outcomes of vaccination with a multiepitope melanoma peptide vaccine plus low-dose interleukin-2 administered either concurrently or on a delayed schedule. J Clin Oncol. 2004 Nov 15; 22(22): 4474-85.
47. Dakappagari NK, Lute KD, Rawale S, et al. Conformational HER-2/neu B-cell epitope peptide vaccine designed to incorporate two native disulfide bonds enhances tumor cell binding and antitumor activities. Biol Chem. 2005 Jan 7; 280(1): 54-63. Epub 2004 Oct 26.
48. Ayyoub M, Mazarguil H, Monsarrat B, et al. A structure - based aproach to designing non - natural peptides that can activate antimelanoma cytotoxic T cells. J Biol Chem, 1999,274:10227.
49. Waeckerle-Men Y, Gander B, Groettrup M. Delivery of tumor antigens to dendritic cells using biodegradable microspheres. Methods Mol Med. 2005; 109:35-46.
50. You Z, Hester J. Rollins L, et al. A Retrogen Strategy for Presentation of an Intracellular Tumor Antigen as an Exogenous Antigen by Dendritic Cells Induces Potent Antitumor T helper and CTL Responses. Cancer Research, 2001,61:197.
51. Tan M C, Mommaas A M, Drijfhout J W, et al. Mannose receptor mediated uptake of antigens strongly enhanced HLA class II restricted antigen presentation by cultured dendritic cells. Eur. J. Immunol, 1999,23:3217.
52. Strome SE, Voss S. Wilcox R, et al. Strategies for antigen loading of dendritic cells to enhance the antitumor immune response. Cancer Res, 2002,62 (6): 1884 ~ 1889.
53. Vieweg J, Jackson A. Modulation of antitumor responses by dendritic cells. Springer Semin Immunopathol. 2005; 26(3): 329-41.
54. Nikitina EY, Chada S, Muro - Cacho C, et al. An effective immunization and cancer treatment with activated dendritic cells transduced with full - length wild - type p53. Gene The, 2002 , 9 (5): 345-352.
55. Trakatelli M, Toungouz M, Lambermont M, et al. Immune characterization of clinical grade-dendritic cells generated from cancer patients and genetically modified by an ALVAC vector carrying MAGE minigenes. Cancer Gene Then 2005; 21:4893-7.
56. Kawada M, Ikeda H, Takahashi T, et al. Vaccination of fusion cells of rat dendritic and carcinoma cells prevents tumor growth in vivo. Int J Cancer. 2003; 105(4): 520-6.
57. Huttner KG, Breuer SK, Paul P, et al. Generation of potent anti-tumor immunity in mice by interleukin-12-secreting dendritic cells. Cancer Immunol Immunother. 2005 Jan; 54(1): 67-77.
58. Lespagnard L, Mettens P, De Smedt T, et al. The immune response induced in vivo by dendritic cells is dependent on B7-1 or B7-2, but the inhibition of both signals does not lead to tolerance. Int Immunol. 1998; 10(3):295-304.
59. Munn DH, Sharma MD, Mellor AL. Ligation of B7-1/B7-2 by human CD4+ T cells triggers indoleamine 2,3-dioxygenase activities in dendritic cells. J Immunol. 2004; 172(7): 4100-10.
60. Zitvogel L, Regnault A, Lozier A, et al. Eradication of established murine tumors using a novel cell-free vaccine: dendritic cell-derived exosomes. Nat Med. 1998; 4(5): 594-600.
61. Amigorena S. Cancer immunotherapy using dendritic cell-derived exosomes. Medicina (B Aires). 2000; 60 Suppl 2:51-4.
62. Morse MA, Garst J, Osada T, et al. A phase I study of dexosome immunotherapy in patients with advanced non-small cell lung cancer. Transl Med. 2005; 3(1):9.
63. Park JH, Kim CJ, Lee JH, et al. Effective immunotherapy of cancer by DNA vaccination. Mol Cells, 1999,9 (4): 384.26
64. De Giovanni C, Nanni P, Forni G. The prospects for cancer gene therapy. Int J Immunopharmacol, 2000,22 (12): 1025.
65. Sumida SM, McKay PF, Truitt DM. Recruitment and expansion of dendritic cells in vivo potentiate the immunogenicity of plasmid DNA vaccines. J Clin Invest. 2004 Nov; 114(9): 1334-42.
66. Zhang S, Li WF, Zhang HJ, et al. Antitumor reactivity of splenocytes primed in vivo with dendritic-cell-based vaccine and secondarily activated with a cocktail of cytokines in vitro. Exp Oncol. 2004; 26(3): 243-5.
67. Coelho M, Gauthier P, Pugniere M, et al. Isolation and characterisation of a human anti-idiotypic scFv used as a surrogate tumour antigen to elicit an anti-HER-2/neu humoral response in mice. Br J Cancer. 2004; 90(10): 2032-41.
68. Malavaud B, Pedron S, Sordello S, et al. Direct FGF receptor 1 activation through an anti-idiotypic strategy mimicks the biological activity of FGF-2 and inhibits the progression of the bladder carcinoma derived from NBT-II cells. Oncogene. 2004; 23(40): 6769-78.
69. Avin E, Haimovich J, Hollander N. Anti-idiotype x anti-CD44 bispecific antibodies inhibit invasion of lymphoid organs by B cell lymphoma. J Immunol. 2004; 173(7): 4736-43.
70. Nicholson S, Bomphray CC, Thomas H, et al. A phase I trial of idiotypic vaccination with HMFG1 in ovarian cancer. Cancer Immunol Immunother. 2004; 53(9):809-16.
71. Binder RJ, Blachere NE, Srivastava PK. Heat shock protein - chaperoned peptides but not free peptides introduced into the cytosol are presented efficiently by major histocompatibility complex I molecules. J Biol Chem, 2001,276 (20) 17163~ 17171.
72. Meng SD, Gao T, Gao GF, Tien P. HBV-specific peptide associated with heat-shock protein gp96. Lancet. 2001; 357(9255): 528-9
73. Srivastava PK. Immunotherapy for human cancer using heat shock protein-Peptide complexes. Curr Oncol Rep. 2005; 7(2): 104-8.
74. Minev B, Hipp J, Firat H, S et al. Cytotoxic T cell immunity against telomerase reverse transcriptase in humans. Proc Natl Acad Sci USA. 2000; 97(9):4796-801.
75. Scappaticci FA, Nolan GP. Induction of anti-tumor immunity in mice using a syngeneic endothelial cell vaccine. Anticancer Res. 2003 Mar-Apr; 23(2B): 1165-72.
76. Susan S, Smyth, Cam Patterson Tiny dancers: the integrin-growth factor nexus in angiogenic signaling. J Cell Biology. 2002; 158(1): 17-21.
77. HAN Quan-Li, DING Jie, FAN Dai-Ming, et al. Selection of mimic epitope of gastric cancer associated antigen by using peptide library. J Fourth Mil Med Univ 2000,21(9) 1162-1164.
78. Liu DW, Tsao YP, Kung Jt, et al. Recombinant adeno-associated virus expressing human papillomavirus type 16 E7 peptide DNA fused with heat shock protein DNA as a potential vaccine for cervical. J Virol. 2000,74(6): 2888-2894.
79. Villen J, de Oliveira E, Nunez JI, et al. Towards a multi-site synthetic vaccine to foot-and-mouth disease: addition of discontinuous site peptide mimic increases the neutralization response in immunized animals Vaccine. 2004 9; 22(27-28): 3523-9.
80. Haicbeur N, Bismuth E, Bosset S, et al. The B subunit of Shiga toxin fused to a tumor antigen elicits CTL and targets dendritic cells to allow MHC class I-restricted presentation of peptides derived from exogenous antigens. J Immunol, 2000, 165(6): 3301-3308.
81. Kate K, Cantwell MJ, Sharma S, et al. Gene transfer of CD40-lig2 and induces autologous immune recognition of chronic lymphocytic leukemia B cells. J Clin Invest, 1998; 5: 1133～1141.
82. Hao X, Shao Y, Ren X, et al. Induction of specific CTL by MAGE-3/CEA peptide-pulsed dendritic cells from HLA-A2/A24(+) gastrointestinal cancer patients. J Cancer Res Clin Oncol. 2002; 128(9): 507-15.
83. Nori, K. D. et al. Tat-conjugated synthetic macromolecules facilitate cytoplasmic drug delivery to human ovarian carcinoma cells. Bioconugate. Chem. 2003, 14, 44-50.
84. Fonseca DP, Frofichs J, Singh M, et al. Induction of antibody and T-cell responses by immunization with ISCOMS containing the 38-kilodalton protein of Mycobacterium tuberculosis[J], Vaccine, 2000, 19(1): 122-131.
85. Brazolot Millan CL, Weeratna R, Krieg AM, et al. CpG DNA can induce strong Thl humeral and cell-mediated immune responses against hepatitis B surface antigen in young mice. Proc Nail Acad Sci USA 1998; 95: 15553-8.
86. Michael J. McCluskie, Risini D, et al. Parenteral and mucosal prime-boost immunization strategies in mice with hepatitis B surface antigen and CpG DNA. FEMS Immunology and Medical Microbiology. 2002; 32: 179-185.
87. Yamamoto S, Yamamoto T, Iho S, et al. Activation of NK cell [human and mouse] by immunostimulatory DNA sequence. Springer Semin Immunopathol. 2000; 22: 35-43.
88. C. Hirunpetcharat, J. Wipasa, S. Sakkhachomphop, et al. CpG oligodeoxynucleotide enhances immunity against blood-stage malaria infection in mice parenterally immunized with a yeast-expressed 19 kDa carboxyl-terminal fragment of Plasmodium yoelii merozoite surface protein-1 [MSP119] formulated in oil-based Muntanides. Vaccine. 2003; 21: 2923-2932.
89. Ningshu Liu, Noriko Ohnishi, Lin Ni, et al. CpG directly induces T-bet expression and inhibits IgG1 and IgE switching in B cells. Nature Immunology. 2003; 4: 687-693.
90. Gramzinski RA, Doolan DL, Sedagah M, et al. Interleukin-12- and gamma interferon-depondent protection against malaria conferred by CpG oligodeoxynucleotide in mice. Infect Immun. 2001; 69: 1643-9.
91. Sparwasser T, Koch ES, Vabulas RM, et al. Bacterial DNA and immunostimulatory CpG oligonucleotides trigger maturation and activation of murine dendritic cells. Eur J Immunol 1998; 28: 2045-54.
92. Wagner H, Hacker H, Lipford GB. Immunostimulatory DNA sequences help to eradicate intracellular pathogens. Springer Semin Immunopathol. 2000; 22: 147-52.
93. Tighe H, Takabayashi K, Schwartz D, et al. Conjugation of immunostimulatory DNA to the short ragweed allergen arab a 1 enhances its immunogenicity and reduces its allergenicity. J Allergy Clin Immunol. 2000; 106: 124-34.
94. Tighe H, Takabayashi K, Schwartz D, et al. Conjugation of protein to immunostimulatory DNA results in a rapid, long-lasting and potent induction of cell-mediated and humoral immunity. Eur J Immunol 2000; 30: 1939-47.
95. Morin C, Barratt G, Fessi H, et al. Improved intracellular deliver of a muramul dipeptide analog by means of nanocapsules. Int J Immunopharmacol. 1994; 16: 451-456.
96. Katare YK, Panda AK, Lalwani K, Haque IU, Ali MM. Potentiation of immune response from polymer-entrapped antigen: toward development of single dose tetanus toxoid vaccine. Drug Deliv. 2003; 10(4): 231-8.
97. Codard G, Boutorrine AS, Saison BE, et al. Antisense effects of cholesterol-oligodeoxynucleotide conjugates associated with poly (alkylcyanoacrylate) nanoparticles. Eur J Biochem. 1995; 232: 404-410.
98. Feng SS, Mu L, Win KY, et al. Nanoparticles of biodegradable polymers for clinical administration of paclitaxel. Curr Med Chem. 2004; 11(4): 413-24.
99. Murthy, N. et al. A macromolecular delivery vehicle for proteinbased vaccines: acid-degradable protein-loaded microgels. Proc. Natl. Acad. Sci. U. S. A. 2003 (100), 4995-5000
100. Desai MP, Labhasetwar V, Amidon GL, et al. Gastrointestinal uptake of biodegradable microparticles: effect of particle size. Pharm Res. 1996; 13(12): 1838-1845.
101. Liu Z, Xiao Y, Chen YH. Epitope-vaccine strategy against HIV-1: today and tomorrow. Immunobiology. 2003; 208(4): 423-8.
102. Markwin P, Velders. Defined flanking spacers and enhanced proteolysis is essential for eradication of established tumors by an epitopesstring DNA vaccine. J Immunol. 2001, 166: 5366-5373.
103. Desmezieres E, Jacob Y, Saron MF. Lyssavirus glycoproteins expressing immunologically potent B cell and. cytotoxic T lymphocyte epitopes as prototypes for multivalent vaccine. J Gene Virol. 1999; 80: 2343.
104. Mireille A, Jean-Pierre K. Recombinant fusion peptides containing single or multiple repeats of a ubiquitous T-helper epitope are highly immunogenic. Molecular Immunology. 1996; 33(13): 1017-1024.
105. Niedermann G, Geier E, Lucchiar/-Hartz M. The specificity of the proteasomes: impact on the MHC class Ⅰ processing and presentation of antigens. Immunol Rev. 1999; 172: 29.
106. Markwin P, Velders, et al. Defined flanking spacers andenhanced proteolysis is essential for eradication ofestablished tumors by an epitope string DNA vaccine. J Immunol. 2001; 166: 5366-5373.
107. Brain L, Claire C, Mark N, et al. A rational strategy to design multiepitope immunogens based on multiple Th lymphocyte epitopes. J Immunol. 2002; 168: 5499-5506.
108. Brain D, Livingston, Mark N, et al. Optomozation of epitope processing enhances immunogenicity of mulfiepitope DNA vaccines. Vaccine. 2001; 19: 4652-4660.
109. Lin Z, Wang Z, Chen YH. Predefined spacers between epitopes on a recombinant