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Cationic Liposomes Loaded with a Synthetic Long Peptide and Poly(I:C): a Defined Adjuvanted Vaccine for Induction of Antigen-Specific T Cell Cytotoxicity

  • Research Article
  • Theme: Nanoparticles in Vaccine Delivery
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Abstract

For effective cancer immunotherapy by vaccination, co-delivery of tumour antigens and adjuvants to dendritic cells and subsequent activation of antigen-specific cytotoxic T cells (CTLs) is crucial. In this study, a synthetic long peptide (SLP) harbouring the model CTL epitope SIINFEKL was encapsulated with the TLR3 ligand poly(inosinic-polycytidylic acid) (poly(I:C)) in cationic liposomes consisting of DOTAP and DOPC. The obtained particles were down-sized to about 140 nm (measured by dynamic light scattering) and had a positive zeta-potential of about 26 mV (according to laser Doppler electrophoresis). SLP loading efficiency was about 40% as determined by HPLC. Poly(I:C) loading efficiency was about 50%, as assessed from the fluorescence intensity of fluorescently labelled poly(I:C). Immunogenicity of the liposomal SLP vaccine was evaluated in vitro by its capacity to activate dendritic cells (DCs) and present the processed SLP to SIINFEKL-specific T cells. The effectiveness of the vaccine to activate CD8+ T cells was analysed in vivo after intradermal and subcutaneous immunisation in mice, by measuring antigen-specific T cells in blood and spleens and assessing their functionality by cytokine production and in vivo cytotoxicity. The liposomal formulation efficiently delivered the SLP to DCs in vitro and induced a functional CD8+ T cell immune response in vivo to the CTL epitope present in the SLP. The SLP-specific CD8+ T cell frequency induced by the poly(I:C)-adjuvanted liposomal SLP formulation showed an at least 25 fold increase over the T cell frequency induced by the poly(I:C)-adjuvanted soluble SLP. In conclusion, cationic liposomes loaded with SLP and poly(I:C) have potential as a powerful therapeutic cancer vaccine formulation.

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References

  1. Melief CJ, van der Burg SH. Immunotherapy of established (pre)malignant disease by synthetic long peptide vaccines. Nat Rev Cancer. 2008;8(5):351–60.

    Article  CAS  PubMed  Google Scholar 

  2. Shanker A, Marincola FM. Cooperativity of adaptive and innate immunity: implications for cancer therapy. Cancer Immunol Immunother. 2011;60(8):1061–74.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  3. Steinman R, Inaba K. Immunogenicity: role of dendritic cells. Bioessays. 1989;10(5):145–52.

    Article  CAS  PubMed  Google Scholar 

  4. Steinman RM. The dendritic cell system and its role in immunogenicity. Annu Rev Immunol. 1991;9:271–96.

    Article  CAS  PubMed  Google Scholar 

  5. Appay V, Douek DC, Price DA. CD8+ T cell efficacy in vaccination and disease. Nat Med. 2008;14(6):623–8.

    Article  CAS  PubMed  Google Scholar 

  6. Vulink A, Radford KJ, Melief C, Hart DN. Dendritic cells in cancer immunotherapy. Adv Cancer Res. 2008;99:363–407.

    Article  CAS  PubMed  Google Scholar 

  7. Inaba K, Young JW, Steinman RM. Direct activation of CD8+ cytotoxic T lymphocytes by dendritic cells. J Exp Med. 1987;166(1):182–94.

    Article  CAS  PubMed  Google Scholar 

  8. Melief CJ. Cancer immunotherapy by dendritic cells. Immunity. 2008;29(3):372–83.

    Article  CAS  PubMed  Google Scholar 

  9. Rosalia RA, Quakkelaar ED, Redeker A, Khan S, Camps M, Drijfhout JW, et al. Dendritic cells process synthetic long peptides better than whole protein, improving antigen presentation and T-cell activation. Eur J Immunol [Research Support, Non-US Gov’t]. 2013;43(10):2554–65.

    CAS  Google Scholar 

  10. Bijker MS, van den Eeden SJ, Franken KL, Melief CJ, van der Burg SH, Offringa R. Superior induction of anti-tumor CTL immunity by extended peptide vaccines involves prolonged, DC-focused antigen presentation. Eur J Immunol. 2008;38(4):1033–42.

    Article  CAS  PubMed  Google Scholar 

  11. Kenter GG, Welters MJ, Valentijn AR, Lowik MJ. Berends-van der Meer DM, Vloon AP, et al. Phase I immunotherapeutic trial with long peptides spanning the E6 and E7 sequences of high-risk human papillomavirus 16 in end-stage cervical cancer patients shows low toxicity and robust immunogenicity. Clin Cancer Res. 2008;14(1):169–77.

    Article  CAS  PubMed  Google Scholar 

  12. Aucouturier J, Ascarateil S, Dupuis L. The use of oil adjuvants in therapeutic vaccines. Vaccine. 2006;24(2):S2-44–5.

    Google Scholar 

  13. Cai S, Yang Q, Bagby TR, Forrest ML. Lymphatic drug delivery using engineered liposomes and solid lipid nanoparticles. Adv Drug Deliv Rev. 2011;63(10–11):901–8.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  14. Zaks K, Jordan M, Guth A, Sellins K, Kedl R, Izzo A, et al. Efficient immunization and cross-priming by vaccine adjuvants containing TLR3 or TLR9 agonists complexed to cationic liposomes. J Immunol. 2006;176(12):7335–45.

    Article  CAS  PubMed  Google Scholar 

  15. Nordly P, Rose F, Christensen D, Nielsen HM, Andersen P, Agger EM, et al. Immunity by formulation design: induction of high CD8+ T-cell responses by poly(I:C) incorporated into the CAF01 adjuvant via a double emulsion method. J Control Release. 2011;150(3):307–17.

    Article  CAS  PubMed  Google Scholar 

  16. Hansen J, Lindenstrom T, Lindberg-Levin J, Aagaard C, Andersen P, Agger EM. CAF05: cationic liposomes that incorporate synthetic cord factor and poly(I:C) induce CTL immunity and reduce tumor burden in mice. Cancer Immunol Immunother. 2012;61(6):893–903.

    Article  CAS  PubMed  Google Scholar 

  17. Gregoriadis G. The immunological adjuvant and vaccine carrier properties of liposomes. J Drug Target. 1994;2(5):351–6.

    Article  CAS  PubMed  Google Scholar 

  18. Al-Jamal WT, Kostarelos K. Liposomes: from a clinically established drug delivery system to a nanoparticle platform for theranostic nanomedicine. Acc Chem Res. 2011;44(10):1094–104.

    Article  CAS  PubMed  Google Scholar 

  19. Hubbell JA, Thomas SN, Swartz MA. Materials engineering for immunomodulation. Nature. 2009;462(7272):449–60.

    Article  CAS  PubMed  Google Scholar 

  20. Watson DS, Endsley AN, Huang L. Design considerations for liposomal vaccines: influence of formulation parameters on antibody and cell-mediated immune responses to liposome associated antigens. Vaccine. 2012;30(13):2256–72.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  21. Bachmann MF, Jennings GT. Vaccine delivery: a matter of size, geometry, kinetics and molecular patterns. Nat Rev Immunol. 2010;10(11):787–96.

    Article  CAS  PubMed  Google Scholar 

  22. Joshi MD, Unger WJ, Storm G, van Kooyk Y, Mastrobattista E. Targeting tumor antigens to dendritic cells using particulate carriers. J Control Release. 2012;161(1):25–37.

    Article  CAS  PubMed  Google Scholar 

  23. Ossendorp F, Mengede E, Camps M, Filius R, Melief CJ. Specific T helper cell requirement for optimal induction of cytotoxic T lymphocytes against major histocompatibility complex class II negative tumors. J Exp Med. 1998;187(5):693–702.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  24. Gregoriadis G, Bacon A, Caparros-Wanderley W, McCormack B. Plasmid DNA vaccines: entrapment into liposomes by dehydration-rehydration. Methods Enzymol. 2003;367:70–80.

    Article  CAS  PubMed  Google Scholar 

  25. Silva AL, Rosalia RA, Sazak A, Carstens MG, Ossendorp F, Oostendorp J, et al. Optimization of encapsulation of a synthetic long peptide in PLGA nanoparticles: low-burst release is crucial for efficient CD8+ T cell activation. Eur J Pharm Biopharm. 2013;83(3):338–45.

    Article  CAS  PubMed  Google Scholar 

  26. Winzler C, Rovere P, Rescigno M, Granucci F, Penna G, Adorini L, et al. Maturation stages of mouse dendritic cells in growth factor-dependent long-term cultures. J Exp Med. 1997;185(2):317–28.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  27. Sanderson S, Shastri N. LacZ inducible, antigen/MHC-specific T cell hybrids. Int Immunol. 1994;6(3):369–76.

    Article  CAS  PubMed  Google Scholar 

  28. Bal SM, Hortensius S, Ding Z, Jiskoot W, Bouwstra JA. Co-encapsulation of antigen and Toll-like receptor ligand in cationic liposomes affects the quality of the immune response in mice after intradermal vaccination. Vaccine. 2011;29(5):1045–52.

    Article  CAS  PubMed  Google Scholar 

  29. Colonna C, Conti B, Genta I, Alpar OH. Non-viral dried powders for respiratory gene delivery prepared by cationic and chitosan loaded liposomes. Int J Pharm. 2008;364(1):108–18.

    Article  CAS  PubMed  Google Scholar 

  30. Voehringer D, Koschella M, Pircher H. Lack of proliferative capacity of human effector and memory T cells expressing killer cell lectinlike receptor G1 (KLRG1). Blood. 2002;100(10):3698–702.

    Article  CAS  PubMed  Google Scholar 

  31. Sallusto F, Geginat J, Lanzavecchia A. Central memory and effector memory T cell subsets: function, generation, and maintenance. Annu Rev Immunol. 2004;22:745–63.

    Article  CAS  PubMed  Google Scholar 

  32. Ma Y, Zhuang Y, Xie X, Wang C, Wang F, Zhou D, et al. The role of surface charge density in cationic liposome-promoted dendritic cell maturation and vaccine-induced immune responses. Nanoscale. 2011;3(5):2307–14.

    Article  CAS  PubMed  Google Scholar 

  33. Chen W, Yan W, Huang L. A simple but effective cancer vaccine consisting of an antigen and a cationic lipid. Cancer Immunol Immunother. 2008;57(4):517–30.

    Article  CAS  PubMed  Google Scholar 

  34. Yan W, Chen W, Huang L. Reactive oxygen species play a central role in the activity of cationic liposome based cancer vaccine. J Control Release. 2008;130(1):22–8.

    Article  CAS  PubMed  Google Scholar 

  35. Campbell RB, Balasubramanian SV, Straubinger RM. Phospholipid-cationic lipid interactions: influences on membrane and vesicle properties. Biochim Biophys Acta. 2001;1512(1):27–39.

    Article  CAS  PubMed  Google Scholar 

  36. de Jong S, Chikh G, Sekirov L, Raney S, Semple S, Klimuk S, et al. Encapsulation in liposomal nanoparticles enhances the immunostimulatory, adjuvant and anti-tumor activity of subcutaneously administered CpG ODN. Cancer Immunol Immunother. 2007;56(8):1251–64.

    Article  PubMed  Google Scholar 

  37. Lesterhuis WJ, de Vries IJ, Schreibelt G, Lambeck AJ, Aarntzen EH, Jacobs JF, et al. Route of administration modulates the induction of dendritic cell vaccine-induced antigen-specific T cells in advanced melanoma patients. Clin Cancer Res. 2011;17(17):5725–35.

    Article  CAS  PubMed  Google Scholar 

  38. Korsholm KS, Hansen J, Karlsen K, Filskov J, Mikkelsen M, Lindenstrom T, et al. Induction of CD8+ T-cell responses against subunit antigens by the novel cationic liposomal CAF09 adjuvant. Vaccine. 2014;32(31):3927–35.

    Article  CAS  PubMed  Google Scholar 

  39. Aguilar JC, Rodriguez EG. Vaccine adjuvants revisited. Vaccine. 2007;25(19):3752–62.

    Article  CAS  PubMed  Google Scholar 

  40. Ludewig B, Barchiesi F, Pericin M, Zinkernagel RM, Hengartner H, Schwendener RA. In vivo antigen loading and activation of dendritic cells via a liposomal peptide vaccine mediates protective antiviral and anti-tumour immunity. Vaccine. 2000;19(1):23–32.

    Article  CAS  PubMed  Google Scholar 

  41. Zhang J, Tian H, Li C, Cheng L, Zhang S, Zhang X, et al. Antitumor effects obtained by autologous Lewis lung cancer cell vaccine engineered to secrete mouse interleukin 27 by means of cationic liposome. Mol Immunol. 2013;55(3–4):264–74.

    Article  CAS  PubMed  Google Scholar 

  42. van Duikeren S, Fransen MF, Redeker A, Wieles B, Platenburg G, Krebber WJ, et al. Vaccine-induced effector-memory CD8+ T cell responses predict therapeutic efficacy against tumors. J Immunol. 2012;189(7):3397–403.

    Article  PubMed  Google Scholar 

  43. van Duikeren S, Arens R. Predicting the efficacy of cancer vaccines by evaluating T-cell responses. Oncoimmunology. 2013;2(1):e22616.

    Article  PubMed Central  PubMed  Google Scholar 

  44. Wang C, Zhuang Y, Zhang Y, Luo Z, Gao N, Li P, et al. Toll-like receptor 3 agonist complexed with cationic liposome augments vaccine-elicited antitumor immunity by enhancing TLR3-IRF3 signaling and type I interferons in dendritic cells. Vaccine. 2012;30(32):4790–9.

    Article  CAS  PubMed  Google Scholar 

  45. Reimer T, Brcic M, Schweizer M, Jungi TW. Poly(I:C) and LPS induce distinct IRF3 and NF-kappaB signaling during type-I IFN and TNF responses in human macrophages. J Leukoc Biol. 2008;83(5):1249–57.

    Article  CAS  PubMed  Google Scholar 

  46. Gitlin L, Barchet W, Gilfillan S, Cella M, Beutler B, Flavell RA, et al. Essential role of mda-5 in type I IFN responses to polyriboinosinic:polyribocytidylic acid and encephalomyocarditis picornavirus. Proc Natl Acad Sci U S A. 2006;103(22):8459–64.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Division of Drug Delivery Technology, Leiden Academic Centre for Drug Research (LACDR), Leiden University, P.O. Box 9502, 2300 RA, Leiden, The Netherlands

    Eleni Maria Varypataki, Koen van der Maaden, Joke Bouwstra & Wim Jiskoot

  2. Department of Immunohematology and Blood Transfusion, Leiden University Medical Centre, P.O. Box 9600, 2300 RC, Leiden, The Netherlands

    Ferry Ossendorp

Authors
  1. Eleni Maria Varypataki
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  2. Koen van der Maaden
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  3. Joke Bouwstra
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  4. Ferry Ossendorp
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  5. Wim Jiskoot
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Correspondence to Ferry Ossendorp or Wim Jiskoot.

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Guest Editor: Aliasger Salem

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Varypataki, E.M., van der Maaden, K., Bouwstra, J. et al. Cationic Liposomes Loaded with a Synthetic Long Peptide and Poly(I:C): a Defined Adjuvanted Vaccine for Induction of Antigen-Specific T Cell Cytotoxicity. AAPS J 17, 216–226 (2015). https://doi.org/10.1208/s12248-014-9686-4

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  • DOI: https://doi.org/10.1208/s12248-014-9686-4

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