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Immunogenicity of Liposomes Containing Lipid Core Peptides and the Adjuvant Quil A

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Purpose

The purpose of this study was to investigate the immunogenicity of liposomes containing mannosylated lipid core peptide (manLCP) constructs, both in vitro and in vivo, with or without the addition of the immune stimulating adjuvant Quil A.

Methods

Mouse bone marrow dendritic cells (BMDC) were cultured with liposome formulations for 48 h, and the resulting level of BMDC activation was determined by flow cytometry. BMDC pulsed with liposome formulations were incubated with 5,6-carboxyfluoroscein diacetate succinimidyl ester-labeled T cells for 72 h and the resulting T cell proliferation was determined by flow cytometry. To investigate the immunogenicity of formulations in vivo, groups of C57Bl/6J mice were immunized by subcutaneous injection, and the resulting antigen-specific cytotoxic and protective immune responses toward tumor challenge evaluated.

Results

Despite being unable to demonstrate the activation of BMDC, BMDC pulsed with liposomes containing manLCP constructs were able to stimulate the proliferation of naïve T cells in vitro. However, in vivo only liposomes containing both manLCP and Quil A were able to stimulate a strong antigen-specific cytotoxic immune response. Liposomes containing manLCP and Quil A within the same particle were able to protect against the growth of tumor cells to a similar level as if the antigen was administered in alum with CD4 help.

Conclusion

ManLCPs administered in liposomes are able to stimulate strong cytotoxic and protective immune responses if Quil A is also incorporated as an adjuvant.

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Abbreviations

BMDC:

bone marrow dendritic cell

DC:

dendritic cell

HEPES:

N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid

ISCOMs:

immune stimulating complexes

LCP:

lipid core peptide

manLCP:

mannose-conjugated LCP

PBS:

phosphate-buffered saline

PC:

l-α-phosphatidylcholine

PRR:

pattern recognition receptor

TEM:

transmission electron microscopy

References

  1. C. Olive M. Batzloff A. Horvath T. Clair P. Yarwood I. Toth M. F. Good (2003) ArticleTitlePotential of lipid core peptide technology as a novel self-adjuvanting vaccine delivery system for multiple different synthetic peptide immunogens Infect. Immun. 71 2373–2383 Occurrence Handle12704107 Occurrence Handle1:CAS:528:DC%2BD3sXjtl2it7o%3D Occurrence Handle10.1128/IAI.71.5.2373-2383.2003

    Article  PubMed  CAS  Google Scholar 

  2. C. Olive M. Batzloff A. Horvath A. Wong T. Clair P. Yarwood I. Toth M. Good (2002) ArticleTitleA lipid core peptide construct containing a conserved region determinant of the group a streptococcal M protein elicits heterologous opsonic antibodies Infect. Immun. 70 2734–2738 Occurrence Handle11953422 Occurrence Handle1:CAS:528:DC%2BD38XjtFKnsb8%3D Occurrence Handle10.1128/IAI.70.5.2734-2738.2002

    Article  PubMed  CAS  Google Scholar 

  3. W. Hayman I. Toth N. Flinn M. Scanlon M. Good (2002) ArticleTitleEnhancing the immunogenicity and modulating the fine epitope recognition of antisera to a helical group A streptococcal peptide vaccine candidate from the M protein using lipid-core peptide technology Immunol. Cell Biol. 80 178–187 Occurrence Handle11940119 Occurrence Handle1:CAS:528:DC%2BD38XjtVGgtLs%3D Occurrence Handle10.1046/j.1440-1711.2002.01067.x

    Article  PubMed  CAS  Google Scholar 

  4. K. White P. Kearns I. Toth S. Hook (2004) ArticleTitleIncreased adjuvant activity of minimal CD8 T cell peptides incorporated into lipid-core-peptides Immunol. Cell Biol. 82 517–522 Occurrence Handle15479437 Occurrence Handle1:CAS:528:DC%2BD2cXhtVShurrP Occurrence Handle10.1111/j.0818-9641.2004.01269.x

    Article  PubMed  CAS  Google Scholar 

  5. Y. Zhan A. Corbett J. Brady R. Sutherland A. Lew (2000) ArticleTitleCD4 help-independent induction of cytotoxic CD8 cells to allogeneic P815 tumor cells is absolutely dependent on co-stimulation J. Immunol. 165 3612–3619 Occurrence Handle11034363 Occurrence Handle1:CAS:528:DC%2BD3cXntVyju7w%3D

    PubMed  CAS  Google Scholar 

  6. B. Wang C. Norbury R. Greenwood J. Bennink J. Yewdell J. Frelinger (2001) ArticleTitleMultiple paths for activation of naive CD8(+) T cells: CD4-independent help J. Immunol. 167 1283–1289 Occurrence Handle11466344 Occurrence Handle1:CAS:528:DC%2BD3MXls1Wkur8%3D

    PubMed  CAS  Google Scholar 

  7. J. Wang A. Livingstone (2003) ArticleTitleCutting edge: CD4(+) T cell help can be essential for primary CD8(+) T cell responses in vivo J. Immunol. 171 6339–6343 Occurrence Handle14662830 Occurrence Handle1:CAS:528:DC%2BD3sXpsV2rs7k%3D

    PubMed  CAS  Google Scholar 

  8. A. Kaiser N. Bercovici J. Abastado A. Nardin (2003) ArticleTitleNaive CD8(+) T cell recruitment and proliferation are dependent on stage of dendritic cell maturation Eur. J. Immunol. 33 162–171 Occurrence Handle12594845 Occurrence Handle1:CAS:528:DC%2BD3sXos1Slsw%3D%3D Occurrence Handle10.1002/immu.200390019

    Article  PubMed  CAS  Google Scholar 

  9. H. Hu H. Winter J. Ma M. Croft W. Urba B. Gox (2002) ArticleTitleCD28, TNF receptor, and IL-12 are critical for CD4-independent cross-priming of therapeutic antitumor CD8(+) T cells J. Immunol. 169 4897–4904 Occurrence Handle12391201

    PubMed  Google Scholar 

  10. H. Guan W. Budzynski R. Koganty M. Krantz M. Reddish J. Rogers B. Longenecker J. Samuel (1998) ArticleTitleLiposomal formulations of synthetic MUC1 peptides: effects of encapsulation versus surface display of peptides on immune responses Bioconjug. Chem. 9 451–458 Occurrence Handle9667946 Occurrence Handle1:CAS:528:DyaK1cXktVanurY%3D Occurrence Handle10.1021/bc970183n

    Article  PubMed  CAS  Google Scholar 

  11. I. Fernandes B. Frisch S. Muller F. Schuber (1997) ArticleTitleSynthetic lipopeptides incorporated in liposomes: in vitro stimulation of the proliferation of murine splenocytes and in vivo induction of an immune response against a peptide antigen Mol. Immunol. 34 569–576 Occurrence Handle9393959 Occurrence Handle1:CAS:528:DyaK2sXnsFSmt7g%3D Occurrence Handle10.1016/S0161-5890(97)00090-4

    Article  PubMed  CAS  Google Scholar 

  12. L. Flebbe H. Braley-Mullen (1986) ArticleTitleImmunopotentiating effects of the adjuvants SGP and Quil A: I. Antibody responses to T-dependent and T-independent antigens Cell. Immunol. 99 119–127 Occurrence Handle3489557 Occurrence Handle1:CAS:528:DyaL28XitFyku74%3D Occurrence Handle10.1016/0008-8749(86)90221-2

    Article  PubMed  CAS  Google Scholar 

  13. S. Kamstrup R. Martin ParticleSan A. Doberti H. Grande K. Dalsgaard (2000) ArticleTitlePreparation and characterisation of Quillaja saponin with less heterogeneity than Quil-A Vaccine 18 2244–2249 Occurrence Handle10717344 Occurrence Handle1:CAS:528:DC%2BD3cXjsFGhu7c%3D Occurrence Handle10.1016/S0264-410X(99)00560-5

    Article  PubMed  CAS  Google Scholar 

  14. M. Baca-Estrada M. Foldvari M. Snider S. Drunen Littel-vanden Hurk Particlevan L. Babiuk (1997) ArticleTitleEffect of IL-4 and IL-12 liposomal formulations on the induction of immune response to bovine herpes virus type-1 glycoprotein D Vaccine 15 1753–1760 Occurrence Handle9364679 Occurrence Handle1:CAS:528:DyaK2sXns1Cktrw%3D Occurrence Handle10.1016/S0264-410X(97)00111-4

    Article  PubMed  CAS  Google Scholar 

  15. A. Ninomiya K. Ogasawara K. Kajino A. Takada H. Kida (2002) ArticleTitleIntranasal administration of a synthetic peptide vaccine encapsulated in liposome together with an anti-CD40 antibody induces protective immunity against influenza A virus in mice Vaccine 20 3123–3129 Occurrence Handle12163263 Occurrence Handle1:CAS:528:DC%2BD38XmtVSrtLY%3D Occurrence Handle10.1016/S0264-410X(02)00261-X

    Article  PubMed  CAS  Google Scholar 

  16. M. Slooten Particlevan G. Storm A. Zoephel Z. Kupcu O. Boerman D. Crommelin E. Wagner R. Kircheis (2000) ArticleTitleLiposomes containing interferon-gamma as adjuvant in tumor cell vaccines Pharm. Res. 17 42–48 Occurrence Handle10714607 Occurrence Handle10.1023/A:1007514424253

    Article  PubMed  Google Scholar 

  17. M. Jaafari M. Foldvari (2002) ArticleTitleTargeting of liposomes to melanoma cells with high levels of ICAM-1 expression through adhesive peptides from immunoglobulin domains J. Pharm. Sci. 91 396–404 Occurrence Handle11835199 Occurrence Handle1:CAS:528:DC%2BD38XhsVSmsrk%3D Occurrence Handle10.1002/jps.10062

    Article  PubMed  CAS  Google Scholar 

  18. M. Copland M. Baird T. Rades J. McKenzie B. Becker F. Reck P. Tyler N. Davies (2003) ArticleTitleLiposomal delivery of antigen to human dendritic cells Vaccine 21 883–890 Occurrence Handle12547598 Occurrence Handle1:CAS:528:DC%2BD3sXltlSgtA%3D%3D Occurrence Handle10.1016/S0264-410X(02)00536-4

    Article  PubMed  CAS  Google Scholar 

  19. G. Kersten D. Crommelin (1995) ArticleTitleLiposomes and ISCOMs as vaccine formulations Biochim. Biophys. Acta 1241 117–138 Occurrence Handle7640293

    PubMed  Google Scholar 

  20. P. Demana N. Davies B. Berger T. Rades (2004) ArticleTitleIncorporation of ovalbumin into ISCOMs and related colloidal particles prepared by the lipid film hydration method Int. J. Pharm. 278 263–274 Occurrence Handle15196631 Occurrence Handle1:CAS:528:DC%2BD2cXkslKmu7o%3D Occurrence Handle10.1016/j.ijpharm.2004.03.021

    Article  PubMed  CAS  Google Scholar 

  21. L. East C. Isacke (2002) ArticleTitleThe mannose receptor family Biochim. Biophys. Acta 1572 364–386 Occurrence Handle12223280 Occurrence Handle1:CAS:528:DC%2BD38XmvVagsbc%3D

    PubMed  CAS  Google Scholar 

  22. F. Sallusto A. Lanzavecchia (1994) ArticleTitleEfficient presentation of soluble antigen by cultured human dendritic cells is maintained by granulocyte/macrophage colony-stimulating factor plus interleukin 4 and down-regulated by tumour necrosis factor α J. Exp. Med. 179 1109–1118 Occurrence Handle8145033 Occurrence Handle1:CAS:528:DyaK2cXhs1Wjurg%3D Occurrence Handle10.1084/jem.179.4.1109

    Article  PubMed  CAS  Google Scholar 

  23. K. Mahnke M. Guo S. Lee H. Sepulveda S. Swain M. Nussenzweg R. Steinman (2000) ArticleTitleThe dendritic cell receptor for endocytosis, DEC-205, can recycle and enhance antigen presentation via major histocompatibility complex class II-positive lysosomal compartments J. Cell Biol. 151 673–683 Occurrence Handle11062267 Occurrence Handle1:CAS:528:DC%2BD3cXnsl2lsb0%3D Occurrence Handle10.1083/jcb.151.3.673

    Article  PubMed  CAS  Google Scholar 

  24. L. Bonifaz D. Bonnyay K. Mahnke M. Rivera M. Nussenzweig R. Steinman (2002) ArticleTitleEfficient targeting of protein antigen to the dendritic cell receptor DEC-205 in the steady state leads to antigen presentation on major histocompatibility complex class I products and peripheral CD8+ T cell tolerance J. Exp. Med. 196 1627–1638 Occurrence Handle12486105 Occurrence Handle1:CAS:528:DC%2BD38XpslWlu7o%3D Occurrence Handle10.1084/jem.20021598

    Article  PubMed  CAS  Google Scholar 

  25. P. Demana C. Fehske K. White T. Rades S. Hook (2004) ArticleTitleEffect of incorporation of the adjuvant Quil A on structure and immune stimulatory capacity of liposomes Immunol. Cell Biol. 82 547–554 Occurrence Handle15479441 Occurrence Handle1:CAS:528:DC%2BD2cXhtVShurrL Occurrence Handle10.1111/j.0818-9641.2004.01276.x

    Article  PubMed  CAS  Google Scholar 

  26. M. Moore F. Carbone M. Bevan (1988) ArticleTitleIntroduction of soluble protein into the class I pathway of antigen processing and presentation Cell 54 777–785 Occurrence Handle3261634 Occurrence Handle1:CAS:528:DyaL1cXmtV2hu74%3D Occurrence Handle10.1016/S0092-8674(88)91043-4

    Article  PubMed  CAS  Google Scholar 

  27. A. Agrawal A. Puri W. Haq C. Gupta (2001) ArticleTitleEnhanced immunostimulant activity and protective effect of a synthetic lipopeptide after liposomization against Plasmodium berghei infection in mice J. Drug Target. 9 231–239 Occurrence Handle11697027 Occurrence Handle1:CAS:528:DC%2BD3MXmt1Sls7o%3D Occurrence Handle10.3109/10611860108997932

    Article  PubMed  CAS  Google Scholar 

  28. R. New (1990) Liposomes: A Practical Approach Oxford University Press New York

    Google Scholar 

  29. M. Steinman D. Hawiger M. C. Nussenzweig (2003) ArticleTitleTolerogenic dendritic cells Annu. Rev. Immunol. 21 685–711 Occurrence Handle12615891 Occurrence Handle1:CAS:528:DC%2BD3sXjtF2is7g%3D Occurrence Handle10.1146/annurev.immunol.21.120601.141040

    Article  PubMed  CAS  Google Scholar 

  30. C. Kensil (1996) ArticleTitleSaponins as vaccine adjuvants Crit. Rev. Ther. Drug Carrier Syst. 13 1–55 Occurrence Handle8853958 Occurrence Handle1:CAS:528:DyaK28XlsV2qt7g%3D

    PubMed  CAS  Google Scholar 

  31. B. Rönnberg M. Fekadu B. Morein (1995) ArticleTitleAdjuvant activity of non-toxic Quillaja saponaria Molina components for use in ISCOM matrix Vaccine 13 1375–1382 Occurrence Handle8585296 Occurrence Handle10.1016/0264-410X(95)00105-A

    Article  PubMed  Google Scholar 

  32. G. Lipford H. Wagner K. Heeg (1994) ArticleTitleVaccination with immunodominant peptides encapsulated in Quil A-containing liposomes induces peptide-specific primary CD8+ cytotoxic T-cells Vaccine 12 73–80 Occurrence Handle8303944 Occurrence Handle1:CAS:528:DyaK2cXit1KlsbY%3D Occurrence Handle10.1016/0264-410X(94)90013-2

    Article  PubMed  CAS  Google Scholar 

  33. S. Behboudi B. Morein M. Villacres-Eriksson (1996) ArticleTitleIn vitro activation of antigen-presenting cells (APC) by defined composition of Quillaja saponaria Molina triterpenoids Clin. Exp. Immunol. 105 26–30 Occurrence Handle8697631 Occurrence Handle1:CAS:528:DyaK28XkvV2ht7k%3D Occurrence Handle10.1046/j.1365-2249.1996.d01-730.x

    Article  PubMed  CAS  Google Scholar 

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Acknowledgment

The authors gratefully acknowledge the Cancer Society of New Zealand for funding for this research.

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Authors and Affiliations

  1. School of Pharmacy, University of Otago, PO Box 913, Dunedin, New Zealand

    Karen White, Thomas Rades & Sarah Hook

  2. School of Pharmacy, University of Queensland, Brisbane, Australia

    Philip Kearns & Istvan Toth

  3. School of Molecular and Microbial Sciences, University of Queensland, Brisbane, Australia

    Istvan Toth

Authors
  1. Karen White
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  2. Thomas Rades
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  3. Philip Kearns
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  4. Istvan Toth
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  5. Sarah Hook
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Correspondence to Sarah Hook.

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White, K., Rades, T., Kearns, P. et al. Immunogenicity of Liposomes Containing Lipid Core Peptides and the Adjuvant Quil A. Pharm Res 23, 1473–1481 (2006). https://doi.org/10.1007/s11095-006-0272-z

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  • DOI: https://doi.org/10.1007/s11095-006-0272-z

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