Molecular Adjuvants Based on Nonpyrogenic Lipophilic Derivatives of norAbuMDP/GMDP Formulated in Nanoliposomes: Stimulation of Innate and Adaptive Immunity
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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.
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.
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.
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 WORDSAdjuvant Bone-marrow radioprotection Liposome Muramyl dipeptide Vaccine
Atomic force microscopy
Antigen presenting cell
Complete Freund’s adjuvant
Dynamic light scattering
1,2-Dioleoyl-sn-Glycero-3-[[N (5-Amino-1-Carboxypentyl) iminodiAcetic Acid] Succinyl] (Nickel Salt)
Granulocyte-monocyte colony-forming cells
Muramyl tripeptide phosphatidylethanolamine
- MT01 – MT08
Lipophilic derivatives based on norAbuMDP and norAbuGMDP
NACHT LRR and PYD domains-containing protein 3
nucleotide-binding oligomerization domain
1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1’-rac-glycerol) (sodium salt)
Recombinant Heat shock protein 90
Size exclusion chromatography
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.
- 3.Takada H, Kotani S. Immunopharmacological activities of synthetic muramyl-peptides. In: Stewart-Tull D, Davier M, editors. Immunology of the bacterial cell envelope. Chichester: Willey; 1985. p. 119–52.Google Scholar
- 7.Lecrec C, Vogel RF. Synthetic immunomodulators and synthetic vaccines. Crit Rev Ther Drug Carrier Syst. 1986;2:353–406.Google Scholar
- 11.No authors listed. Mifamurtide: osteosarcoma: ineffective and harmful. Prescrire Int. 2011;20:89Google Scholar
- 15.Krupka M, Masek J, Bartheldyova E, Turanek Knotigova P, Plockova J, Korvasova Z, et al. Enhancement of immune response towards non-lipidized Borrelia burgdorferi recombinant OspC antigen by binding onto the surface of metallochelating nanoliposomes with entrapped lipophilic derivatives of norAbuMDP. J Control Release. 2012;160(2):374–81.CrossRefPubMedGoogle Scholar
- 20.Masek J, Bartheldyova E, Turanek-Knotigova P, Skrabalova M, Korvasova Z, Plockova J, et al. Metallochelating liposomes with associated lipophilised norAbuMDP as biocompatible platform for construction of vaccines with recombinant His-tagged antigens: preparation, structural study and immune response towards rHsp90. J Control Release. 2011;151(2):193–201.CrossRefPubMedGoogle Scholar
- 26.Sakamoto S, Okawa T, Ogawa N. Therapeutic effect of microtasin on cancer patients with leucopenia during radiation therapy. J New Rem Clin. 1989;39:1407–22.Google Scholar
- 43.Turanek J, Kasna A, Koudela B, Ledvina M, Miller AD. Stimulation of innate immunity in newborn kids against Cryptosporidium parvum infection-challenge by intranasal/per-oral administration of liposomal formulation of N-L 18-norAbu-GDMP adjuvant. Parasitology. 2005;131:601–8.CrossRefPubMedGoogle Scholar