title: Combination of MIDGE-Th1 DNA vaccines with the cationic lipid SAINT-18: studies on formulation, biodistribution and vector clearance.
abstract: We have previously shown that the combination of MIDGE-Th1 DNA vectors with the cationic lipid SAINT-18 increases the immune response to the encoded antigen in mice. Here, we report on experiments to further optimize and characterize this approach. We evaluated different formulations of MIDGE-Th1 vectors with SAINT-18 by assessing their influence on the transfection efficiency in cell culture and on the immune response in mice. We found that high amounts of SAINT-18 in formulations with a w/w ratio MIDGE Th1/SAINT-18 of 1:4.8 are beneficial for cell transfection in vitro. In contrast, the formulation of HBsAg-encoding MIDGE-Th1 DNA vectors with the lowest amount of SAINT-18 (w/w ratio MIDGE Th1/SAINT-18 of 1:0.5) resulted in the highest serum IgG1 and IgG2a levels after intradermal immunization of mice. Consequently, latter formulation was selected for a comparative biodistribution study in rats. Following intradermal administration of both naked and formulated MIDGE-Th1 DNA, the vectors localized primarily at the site of injection. Vector DNA levels decreased substantially over the two months duration of the study. When administered in combination with SAINT-18, the vectors were found in significantly higher amounts in draining lymph nodes in comparison to administration of naked MIDGE-Th1 DNA. We propose that the high immune responses induced by MIDGE-Th1/SAINT-18 lipoplexes are mediated by enhanced transfection of cells in vivo, resulting in stronger antigen expression and presentation. Importantly, the combination of MIDGE-Th1 vectors with SAINT-18 was well tolerated in mice and rats and is expected to be safe in human clinical applications.
authors.affiliation: MOLOGEN AG, Fabeckstraße 30, 14195 Berlin, Germany. Electronic address: endmann@mologen.com.
authors.forename: Anne
authors.initials: A
authors.lastname: Endmann
authors.affiliation: MOLOGEN AG, Fabeckstraße 30, 14195 Berlin, Germany.
authors.forename: Detlef
authors.initials: D
authors.lastname: Oswald
authors.affiliation: MOLOGEN AG, Fabeckstraße 30, 14195 Berlin, Germany.
authors.forename: Oliver
authors.initials: O
authors.lastname: Riede
authors.affiliation: Synvolux Therapeutics B.V., L.J. Zielstraweg 1, 9713 GX Groningen, The Netherlands.
authors.forename: Eduard G
authors.initials: EG
authors.lastname: Talman
authors.affiliation: Synvolux Therapeutics B.V., L.J. Zielstraweg 1, 9713 GX Groningen, The Netherlands.
authors.forename: Roelien E
authors.initials: RE
authors.lastname: Vos
authors.affiliation: MOLOGEN AG, Fabeckstraße 30, 14195 Berlin, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Schroff
authors.affiliation: MOLOGEN AG, Fabeckstraße 30, 14195 Berlin, Germany.
authors.forename: Christiane
authors.initials: C
authors.lastname: Kleuss
authors.affiliation: Synvolux Therapeutics B.V., L.J. Zielstraweg 1, 9713 GX Groningen, The Netherlands; Department of Pathology and Medical Biology, University Medical Center Groningen, University of Groningen, Hanzeplein 1, 9713 GZ Groningen, The Netherlands.
authors.forename: Marcel H J
authors.initials: MH
authors.lastname: Ruiters
authors.affiliation: MOLOGEN AG, Fabeckstraße 30, 14195 Berlin, Germany.
authors.forename: Christiane
authors.initials: C
authors.lastname: Juhls
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D002412:Cations; D005260:Female; D005822:Genetic Vectors / Q000737:chemistry* / Q000493:pharmacokinetics; D006514:Hepatitis B Surface Antigens / Q000276:immunology; D007074:Immunoglobulin G / Q000097:blood; D008198:Lymph Nodes / Q000276:immunology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D011726:Pyridinium Compounds / Q000737:chemistry* / Q000493:pharmacokinetics; D051381:Rats; D017208:Rats, Wistar; D018417:Th1 Cells / Q000276:immunology; D014018:Tissue Distribution; D014162:Transfection; D019444:Vaccines, DNA / Q000276:immunology* / Q000493:pharmacokinetics
chemical_list: C532953:1-methyl-4-(9-dioleyl)methylpyridinium; D000914:Antibodies, Viral; D002412:Cations; D006514:Hepatitis B Surface Antigens; D007074:Immunoglobulin G; D011726:Pyridinium Compounds; D019444:Vaccines, DNA
keywords: Biodistribution; Cationic lipid; DNA vaccine; MIDGE-Th1; SAINT-18; Transfection reagent