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Pharmaceutical Research

, Volume 32, Issue 4, pp 1186–1199 | Cite as

Molecular Adjuvants Based on Nonpyrogenic Lipophilic Derivatives of norAbuMDP/GMDP Formulated in Nanoliposomes: Stimulation of Innate and Adaptive Immunity

  • Pavlína Turánek Knotigová
  • Daniel Zyka
  • Josef Mašek
  • Anna Kovalová
  • Michal Křupka
  • Eliška Bartheldyová
  • Pavel Kulich
  • Štěpán Koudelka
  • Róbert Lukáč
  • Zuzana Kauerová
  • Antonín Vacek
  • Milada Stuchlová Horynová
  • Alois Kozubík
  • Andrew D. Miller
  • Ladislav Fekete
  • Irena Kratochvílová
  • Jan Ježek
  • Miroslav Ledvina
  • Milan Raška
  • Jaroslav Turánek
Research Paper

ABSTRACT

Purpose

The aim of this work was to demonstrate an immunostimulatory and adjuvant effect of new apyrogenic lipophilic derivatives of norAbuMDP and norAbuGMDP formulated in nanoliposomes.

Methods

Nanoliposomes and metallochelating nanoliposomes were prepared by lipid film hydration and extrusion methods. The structure of the liposomal formulation was studied by electron microscopy, AF microscopy, and dynamic light scattering. Sublethal and lethal γ-irradiation mice models were used to demonstrate stimulation of innate immune system. Recombinant Hsp90 antigen (Candida albicans) bound onto metallochelating nanoliposomes was used for immunisation of mice to demonstrate adjuvant activities of tested compounds.

Results

Safety and stimulation of innate and adaptive immunity were demonstrated on rabbits and mice. The liposomal formulation of norAbuMDP/GMDP was apyrogenic in rabbit test and lacking any side effect in vivo. Recovery of bone marrow after sublethal γ-irradiation as well as increased survival of mice after lethal irradiation was demonstrated. Enhancement of specific immune response was demonstrated for some derivatives incorporated in metallochelating nanoliposomes with recombinant Hsp90 protein antigen.

Conclusions

Liposomal formulations of new lipophilic derivatives of norAbuMDP/GMDP proved themselves as promising adjuvants for recombinant vaccines as well as immunomodulators for stimulation of innate immunity and bone-marrow recovery after chemo/radio therapy of cancer.

KEY WORDS

Adjuvant Bone-marrow radioprotection Liposome Muramyl dipeptide Vaccine 

ABBREVIATIONS

AFM

Atomic force microscopy

APC

Antigen presenting cell

CFA

Complete Freund’s adjuvant

CMI

Cell-mediated immunity

DLS

Dynamic light scattering

DOGS-NTA-Ni

1,2-Dioleoyl-sn-Glycero-3-[[N (5-Amino-1-Carboxypentyl) iminodiAcetic Acid] Succinyl] (Nickel Salt)

EM

Electron microscopy

EPC

Egg phosphatidylcholine

GM-CFC

Granulocyte-monocyte colony-forming cells

iE-DAP

γ-D-glutamyl-meso-diaminopimelic acid

MDP

Muramyl dipeptide

MTP-PE

Muramyl tripeptide phosphatidylethanolamine

MT01 – MT08

Lipophilic derivatives based on norAbuMDP and norAbuGMDP

NALP3

NACHT LRR and PYD domains-containing protein 3

NOD

nucleotide-binding oligomerization domain

norAbuMDP

NorMurNAc-L-Abu-D-iGln

norAbuGMDP

NorAbu-glucosaminylmuramyl dipeptide

POPG

1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1’-rac-glycerol) (sodium salt)

rHsp90

Recombinant Heat shock protein 90

SEC

Size exclusion chromatography

TLR

Toll-like receptor

Notes

ACKNOWLEDGMENTS AND DISCLOSURES

By this paper we would like to revere the memory of deceased MUDr. Antonín Vacek, CSc., who pioneered the field of the effect of γ-rays on the immune system and haematopoiesis. His scientific work contributed to the cosmic flight of spaceships with human crew and was awarded by NASA and Russian space agency within the programme Intercosmos. Moreover, we would like to revere the memory of Prof. Antonín Holý, deceased in 2012, who pioneered the field of modern antiviral drugs. The development of new norAbuMDP/GMDP analogues would not have been possible without his long-lasting support to this project.

This Work was Supported by Grants

GAP304/10/1951 to J.T, M.L, M.R; TAČR TA01011165 J.T, M.L, M.R ; GAP503/12/G147 to J.T; FNUSA-ICRC CZ.1.07/2.3.00/30.0043; The Ministry of Education, Youth and Sports of the Czech Republic (CZ.1.07/2.3.00/20.0164) to J.T. and M.R.; LF_2014_020 to M.K. and M.R., the projects MZE0002716202 of the Czech Ministry of Agriculture. Special thanks to the project AdmireVet (CZ.1.05/2.1.00/01.0006 - ED0006/01/01 from the Czech Ministry of Education) for permit the access to the scanning electron microscope.

Conflict of Interest

The authors have declared that no conflict of interest exists. I certify that this manuscript, or any part of it, has not been published and will not be submitted elsewhere for publication while being considered by the journal Pharmaceutical Research.

Supplementary material

11095_2014_1516_MOESM1_ESM.pdf (616 kb)
ESM 1 (PDF 616 kb)

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Pavlína Turánek Knotigová
    • 1
  • Daniel Zyka
    • 2
  • Josef Mašek
    • 1
  • Anna Kovalová
    • 2
  • Michal Křupka
    • 3
  • Eliška Bartheldyová
    • 1
  • Pavel Kulich
    • 1
  • Štěpán Koudelka
    • 1
    • 8
  • Róbert Lukáč
    • 1
  • Zuzana Kauerová
    • 1
  • Antonín Vacek
    • 1
  • Milada Stuchlová Horynová
    • 3
  • Alois Kozubík
    • 4
  • Andrew D. Miller
    • 5
  • Ladislav Fekete
    • 6
  • Irena Kratochvílová
    • 6
  • Jan Ježek
    • 2
  • Miroslav Ledvina
    • 7
  • Milan Raška
    • 3
  • Jaroslav Turánek
    • 1
  1. 1.Department of Pharmacology and ImmunotherapyVeterinary Research InstituteBrnoCzech Republic
  2. 2.Institute of Organic Chemistry and BiochemistryCzech Academy of SciencesPragueCzech Republic
  3. 3.Faculty of Medicine and Dentistry, Department of ImmunologyPalacky University Olomouc,OlomoucCzech Republic
  4. 4.Institute of BiophysicsAcademy of Sciences of the Czech Republic, v.v.iBrnoCzech Republic
  5. 5.Pharmaceutical Sciences DivisionKing’s College LondonLondonUK
  6. 6.Institute of PhysicsCzech Academy of SciencesPragueCzech Republic
  7. 7.Department of Chemistry of Natural CompoundsInstitute of Chemical TechnologyPrague 6Czech Republic
  8. 8.International Research CenterSt. Anne’s University Hospital BrnoBrnoCzech Republic

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