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EphA2-derived peptide vaccine with amphiphilic poly(γ-glutamic acid) nanoparticles elicits an anti-tumor effect against mouse liver tumor

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Abstract

The prognosis of liver cancer remains poor, but recent advances in nanotechnology offer promising possibilities for cancer treatment. Novel adjuvant, amphiphilic nanoparticles (NPs) composed of l-phenylalanine (Phe)-conjugated poly(γ-glutamic acid) (γ-PGA-Phe NPs) having excellent capacity for carrying peptides, were found to have the potential for use as a peptide vaccine against tumor models overexpressing artificial antigens, such as ovalbumin (OVA). However, the anti-tumor potential of γ-PGA-Phe NPs vaccines using much less immunogenic tumor-associated antigen (TAA)-derived peptide needs to be clarified. In this study, we evaluated the effectiveness of immunization with EphA2, recently identified TAA, derived peptide-immobilized γ-PGA-Phe NPs (Eph-NPs) against mouse liver tumor of MC38 cells (EphA2-positive colon cancer cells). Immunization of normal mice with Eph-NPs resulted in generation of EphA2-specific type-1 CD8+ T cells. Immunization with Eph-NPs tended to provide a degree of anti-MC38 liver tumor protection more than that observed for immunization with the mixture of EphA2-derived peptide and complete Freund’s adjuvant (Eph + CFA). Neither Eph-NPs nor Eph + CFA vaccines inhibited tumor growth of BL6, EphA2-negative melanoma cells. Splenocytes isolated from MC38-bearing mice treated with Eph-NPs showed strong and specific cytotoxic activity against MC38 cells. Immunization with Eph + CFA induced liver damage as evidenced by elevation of serum alanine aminotransferase, while Eph-NPs vaccination did not exhibit any toxic damage to the liver. These results demonstrated that immunization with Eph-NPs displayed anti-tumor effects against liver tumor by generating acquired immunity equivalent to the toxic adjuvant CFA, suggesting that safe γ-PGA-Phe NPs could be applied clinically for the vaccine treatment of liver cancer.

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Abbreviations

IFA:

Incomplete Freund’s adjuvant

NPs:

Nanoparticles

γ-PGA:

Poly(γ-glutamic acid)

Phe:

l-Phenylalanine

CFA:

Complete Freund’s adjuvant

PBS:

Phosphate buffered saline

i.p.:

Intraperitoneal

ALT:

Alanine aminotransferase

DCs:

Dendritic cells

References

  1. Berzofsky JA, Ahlers JD, Belyakov IM (2001) Strategies for designing and optimizing new generation vaccines. Nat Rev Immunol 1:209–219

    Article  PubMed  CAS  Google Scholar 

  2. Schijns VE (2000) Immunological concepts of vaccine adjuvant activity. Curr Opin Immunol 12:456–463

    Article  PubMed  CAS  Google Scholar 

  3. Valmori D, Souleimanian NE, Tosello V et al (2007) Vaccination with NY-ESO-1 protein and CPG in Montanide induces integrated antibody/Th1 responses and CD8 T cells through cross-priming. Proc Natl Acad Sci USA 104:8947–8952

    Article  PubMed  CAS  Google Scholar 

  4. Wang F, Bade E, Kuniyoshi C et al (1999) Phase 1 trial of a MART-1 peptide vaccine with incomplete Freund’s adjuvants for resected high-risk melanoma. Clin Cancer Res 5:2756–2765

    PubMed  CAS  Google Scholar 

  5. Gilewski TA, Ragupathi G, Dickler M et al (2007) Immunization of high-risk breast cancer patients with clustered sTn-KLH conjugate plus the immunologic adjuvant QS-21. Clin Cancer Res 13:2977–2985

    Article  PubMed  CAS  Google Scholar 

  6. Bottomley A, Debruyne C, Ferip E et al (2008) Symptom and quality of life results of an international randomized phase 3 study of adjuvant vaccination with Bec2/BCG in responding patients with limited disease small-cell lung cancer. Eur J Cancer 44:2178–2184

    Article  PubMed  CAS  Google Scholar 

  7. Perales MA, Yuan J, Powel S et al (2008) Phase 1/2 study of GM-CSF DNA as an adjuvant for a multipeptide cancer vaccine in patients with advanced melanoma. Mol Ther 16:2022–2029

    Article  PubMed  CAS  Google Scholar 

  8. Mocellin S, Riccardo-Rossi C, Lise M et al (2004) Colorectal cancer vaccines: principles, results, and perspectives. Gastroenterology 27:1821–1837

    Article  CAS  Google Scholar 

  9. Dinauer N, Balthasar S, Weber C et al (2005) Selective targeting of antibody-conjugated nanoparticles to leukemic cells and primary T-lymphocytes. Biomaterials 26:5898–5906

    Article  PubMed  CAS  Google Scholar 

  10. Khatri K, Goyal AK, Gupta N et al (2008) Plasmid DNA loaded chitosan nanoparticles for nasal mucosal immunization against hepatitis B. Int J Pharm 354:235–241

    PubMed  CAS  Google Scholar 

  11. Almeida AJ, Souto E (2007) Solid lipid nanoparticles as a drug delivery system for peptides and proteins. Adv Drug Deliv Rev 59:478–490

    Article  PubMed  CAS  Google Scholar 

  12. Hayakawa T, Kawamura M, Okamoto M et al (1998) Concanavalin A-immobilized polystyrene nanospheres capture HIV-1 and gp120: potential approach towards prevention of viral transmission. J Med Virol 56:327–331

    Article  PubMed  CAS  Google Scholar 

  13. Kawamura M, Naito T, Ueno M et al (2002) Induction of mucosal IgA following intravaginal administration of inactivated HIV-1-capturing nanospheres in mice. J Med Virol 66:291–298

    Article  PubMed  CAS  Google Scholar 

  14. Akagi T, Kawamura M, Ueno M et al (2003) Mucosal immunization with inactivated HIV-1-capturing nanospheres induces a significant HIV-1-specific vaginal antibody response in mice. J Med Virol 69:163–172

    Article  PubMed  Google Scholar 

  15. Miyake A, Akagi T, Enose Y et al (2004) Induction of HIV-specific antibody response and protection against vaginal SHIV transmission by intranasal immunization with inactivated SHIV-capturing nanospheres in macaques. J Med Virol 73:368–377

    Article  PubMed  CAS  Google Scholar 

  16. Akagi T, Wang X, Uto T, Baba M, Akashi M (2007) Protein direct delivery to dendritic cells using nanoparticles based on amphiphilic poly(amino acid) derivatives. Biomaterials 28:3427–3436

    Article  PubMed  CAS  Google Scholar 

  17. Shih IL, Van YT (2001) The production of poly-(γ-glutamic acid) from microorganisms and its various applications. Bioresour Technol 79:207–225

    Article  PubMed  CAS  Google Scholar 

  18. Obst M, Steinbüchel A (2004) Microbial degradation of poly(amino acid)s. Biomacromolecules 5:1166–1176

    Article  PubMed  CAS  Google Scholar 

  19. Sung MH, Park C, Kim CJ, Poo H, Soda K, Ashiuchi M (2005) Natural and edible biopolymer poly-γ-glutamic acid: synthesis, production, and applications. Chem Rec 5:352–366

    Article  PubMed  CAS  Google Scholar 

  20. Akagi T, Higashi M, Kaneko T et al (2005) In vitro enzymatic degradation of nanoparticles prepared from hydrophobically-modified poly(γ-glutamic acid). Macromol Biosci 14:598–602

    Article  CAS  Google Scholar 

  21. Akagi T, Kaneko T, Kida T, Akashi M (2006) Multifunctional conjugation of proteins on/into bio-nanoparticles prepared by amphiphilic poly(γ-glutamic acid). J Biomater Sci Polym Ed 17:875–892

    Article  PubMed  CAS  Google Scholar 

  22. Uto T, Wang X, Sato K et al (2007) Targeting of antigen to dendritic cells with poly(gamma-glutamic acid) nanoparticles induces antigen-specific humoral and cellular immunity. J Immunol 178:2979–2986

    PubMed  CAS  Google Scholar 

  23. Olson RM, Perencevich NP, Malcolm AW et al (1980) Patterns of recurrence following curative resection of adenocarcinoma of the colon and rectum. Cancer 45:2969–2974

    Article  PubMed  CAS  Google Scholar 

  24. Malcolm AW, Perencevich NP, Olson RM et al (1981) Analysis of recurrence patterns following curative resection for carcinoma of the colon and rectum. Surg Gynecol Obstet 152:131–136

    PubMed  CAS  Google Scholar 

  25. Yamaguchi S, Tatsumi T, Takehara T et al (2007) Immunotherapy of murine colon cancer using receptor tyrosine kinase EphA2-derived peptide-pulsed dendritic cell vaccines. Cancer 110:1469–1477

    Article  PubMed  CAS  Google Scholar 

  26. Yamaguchi S, Tatsumi T, Takehara T et al (2008) Dendritic cell-based vaccines suppress metastatic liver tumor via activation of local innate and acquired immunity. Cancer Immunol Immunother 57:1861–1869

    Article  PubMed  CAS  Google Scholar 

  27. Akagi T, Kaneko T, Kida T et al (2005) Preparation and characterization of biodegradable nanoparticles based on poly(γ-glutamic acid) with l-phenylalanine as a protein carrier. J Control Release 108:226–236

    Article  PubMed  CAS  Google Scholar 

  28. Tatsumi T, Gambotto A, Robbins PD et al (2002) Interleukin 18-gene transfer expands the repertoire of anti-tumor Th1-type immunity elicited by dendritic cell- based vaccines in association with enhanced therapeutic efficacy. Cancer Res 62:5853–5858

    PubMed  CAS  Google Scholar 

  29. Yoshikawa T, Okada N, Oda A et al (2008) Development of amphiphilic γ-PGA-nanoparticle based tumor vaccine: potential of the nanoparticle cytosolic protein delivery carrier. Biochem Biophys Res Commun 366:408–413

    Article  PubMed  CAS  Google Scholar 

  30. Yoshikawa T, Okada N, Oda A et al (2008) Nanoparticles built by self-assembly of amphiphilic γ-PGA can deliver antigens to antigen-presenting cells with high efficiency: a new tumor-vaccine carrier for eliciting effector T cells. Vaccine 26:1303–1313

    Article  PubMed  CAS  Google Scholar 

  31. Uto T, Wang X, Akagi T et al (2009) Improvement of adaptive immunity by antigen-carrying biodegradable nanoparticles. Biochem Biophys Res Commun 379:600–604413

    Article  PubMed  CAS  Google Scholar 

  32. Dhodapkar MV, Young JW, Chapman PB et al (2006) Paucity of functional T-cell memory to melanoma antigens in healthy donors and melanoma patients. Clin Cancer Res 6:4831–4838

    Google Scholar 

  33. Zantek ND, Azimi M, Fedor-Chaiken M, Wang B, Brackenbury R, Kinch MS (1999) E-cadherin regulates the function of the EphA2 receptor tyrosine kinase. Cell Growth Differ 10:629–638

    PubMed  CAS  Google Scholar 

  34. DeRisi J, Penland L, Brown PO et al (1996) Use of a cDNA microarray to analyse gene expression patterns in human cancer. Nat Genet 14:457–460

    Article  PubMed  CAS  Google Scholar 

  35. Easty DJ, Hill SP, Hsu MY et al (1999) Up-regulation of ephrin-A1 during melanoma progression. Int J Cancer 84:494–501

    Article  PubMed  CAS  Google Scholar 

  36. Lu M, Miller KD, Gokmen-Polar Y, Jeng MH, Kinch MS (2003) EphA2 overexpression decreases estrogen dependence and tamoxifen sensitivity. Cancer Res 63:3425–3429

    PubMed  CAS  Google Scholar 

  37. Zelinski DP, Zantek ND, Stewart JC, Irizarry AR, Kinch MS (2001) EphA2 overexpression causes tumorigenesis of mammary epithelial cells. Cancer Res 61:2301–2306

    PubMed  CAS  Google Scholar 

  38. Branan JM, Dong W, Prudkin L et al (2009) Expression of the receptor tyrosine kinase EphA2 is increased in smokers and predicts poor survival in non-small cell lung cancer. Clin Cancer Res 15:4423–4430

    Article  CAS  Google Scholar 

  39. Duxbury MS, Ito H, Zinner MJ, Ashley SW, Whang EE (2004) EphA2: a determinant of malignant cellular behavior and a potential therapeutic target in pancreatic adenocarcinoma. Oncogene 23:1448–1456

    Article  PubMed  CAS  Google Scholar 

  40. D’Amico TA, Aloia TA, Moore MB et al (2001) Predicting the sites of metastases from lung cancer using molecular biologic markers. Ann Thorac Surg 72:1144–1148

    Article  PubMed  Google Scholar 

  41. Broderson JR (1989) A retrospective review of lesions associated with the use of Freund’s adjuvant. Lab Anim Sci 39:400–405

    PubMed  CAS  Google Scholar 

  42. Amyx HL (1987) Control of animal pain and distress in antibody production and infectious disease studies. J Am Vet Med Assoc 191:1287–1289

    PubMed  CAS  Google Scholar 

  43. Toth LA, Dunlap AW, Olson GA et al (1989) An evaluation of distress following intraperitoneal immunization with Freund’ adjuvant in mice. Lab Anim Sci 39:122–126

    PubMed  CAS  Google Scholar 

  44. George AP, Catherine AP (2005) Liver immunobiology. Toxicol Pathol 33:52–62

    Article  CAS  Google Scholar 

  45. Doherty DG, O’Farrelly C (2000) Innate and adaptive lymphoid cells in human liver. Immunol Rev 174:5–20

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by a Grant-in-Aid from Core Research for Evolutional Science and Technology (CREST) from Japan Science and Technology Agency (JST).

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Authors and Affiliations

  1. Department of Gastroenterology and Hepatology, Osaka University Graduate School of Medicine, 2-2 Yamadaoka, Suita, Osaka, 565-0871, Japan

    Shinjiro Yamaguchi, Tomohide Tatsumi, Tetsuo Takehara, Akira Sasakawa, Masashi Yamamoto, Keisuke Kohga, Takuya Miyagi, Tatsuya Kanto, Naoki Hiramastu & Norio Hayashi

  2. Department of Applied Chemistry, Graduate School of Engineering, Osaka University, Suita, Japan

    Takami Akagi & Mitsuru Akashi

  3. Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST), Tokyo, Japan

    Shinjiro Yamaguchi, Tomohide Tatsumi, Tetsuo Takehara, Akira Sasakawa, Masashi Yamamoto, Takami Akagi, Mitsuru Akashi & Norio Hayashi

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  1. Shinjiro Yamaguchi
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  2. Tomohide Tatsumi
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  3. Tetsuo Takehara
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  10. Takami Akagi
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Corresponding author

Correspondence to Tomohide Tatsumi.

Additional information

S. Yamaguchi and T. Tatsumi contributed equally to this work.

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Yamaguchi, S., Tatsumi, T., Takehara, T. et al. EphA2-derived peptide vaccine with amphiphilic poly(γ-glutamic acid) nanoparticles elicits an anti-tumor effect against mouse liver tumor. Cancer Immunol Immunother 59, 759–767 (2010). https://doi.org/10.1007/s00262-009-0796-2

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