title: Liposomes containing glucosyl ceramide specifically bind T4 bacteriophage: a self-assembling nanocarrier formulation.
abstract: A unique formulation is described comprising liposomes containing glucosyl ceramide (GluCer) in the lipid bilayer to which bacteriophage T4 was attached. Binding of the phage T4 did not occur to glycolipids, such as galactosyl ceramide, containing an aldose in which the C-2 or C-4 conformations were not identical to glucose. These results strongly support previous proposals that glucose is a major receptor moiety for T4 binding to Escherichia coli. By using the binding of T4 to liposomal GluCer, we further describe a formulation that can be used as a self-assembling combined antigen and adjuvant carrier. A peptide antigen derived from C-trimer (Ct) of HIV-1 gp41 was fused to the highly antigenic outer capsid protein (Hoc), a nonessential protein of T4 that spontaneously binds to the T4 capsid. This resulted in display of the Ct-Hoc construct on the T4 capsid, and specific binding of a human monoclonal antibody that recognizes a peptide sequence of Ct was demonstrated. Liposomes containing monophosphoryl lipid A (MPLA) have been demonstrated to have potent adjuvant activities for experimental vaccines both in humans and animals, and because of this, mice were immunized with the Ct-Hoc-T4 construct that was bound to liposomes containing both GluCer and MPLA, resulting in the induction of high titers of Ct-specific antibodies. We conclude that liposomes containing both GluCer and MPLA can spontaneously bind to a construct of T4 that displays antigens that spontaneously binds to the capsid of T4 bacteriophage. This formulation could be utilized as an easily manufactured self-assembling antigen and adjuvant carrier.
authors.affiliation: Department of Adjuvant and Antigen Research, U.S. Military HIV Research Program, Division of Retrovirology, Walter Reed Army Institute of Research, Rockville, Maryland, USA. calving@hivresearch.org
authors.forename: Carl R
authors.initials: CR
authors.lastname: Alving
authors.forename: Mangala
authors.initials: M
authors.lastname: Rao
authors.forename: Kristina K
authors.initials: KK
authors.lastname: Peachman
authors.forename: Ludmila
authors.initials: L
authors.lastname: Asher
authors.forename: Venigalla B
authors.initials: VB
authors.lastname: Rao
mesh_terms: D000276:Adjuvants, Immunologic / Q000737:chemistry; D000595:Amino Acid Sequence; D000818:Animals; D017122:Bacteriophage T4 / Q000378:metabolism* / Q000648:ultrastructure; D005963:Glucosylceramides / Q000737:chemistry*; D015488:HIV Antigens / Q000235:genetics / Q000276:immunology; D015497:HIV-1 / Q000276:immunology; D006801:Humans; D008050:Lipid A / Q000031:analogs & derivatives / Q000737:chemistry; D008051:Lipid Bilayers / Q000737:chemistry; D008081:Liposomes / Q000737:chemistry* / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D049329:Nanostructures / Q000737:chemistry*
chemical_list: D000276:Adjuvants, Immunologic; D005963:Glucosylceramides; D015488:HIV Antigens; D008050:Lipid A; D008051:Lipid Bilayers; D008081:Liposomes; C048436:monophosphoryl lipid A