title: Synthesis of Mannosylated Lipopeptides with Receptor Targeting Properties.
abstract: Present on the surface of antigen presenting cells (APCs), the mannose receptor (MR) has long been recognized as a front-line receptor in pathogen recognition. During the past decade many attempts have been made to target this receptor for applications including vaccine and drug development. In the present study, a library of vaccine constructs comprising fluorescently labeled mannosylated lipid-dendrimers that contained the ovalbumin CD4(+) epitope, OVA(323-339), as the model peptide antigen were synthesized using fluorenylmethyloxycarbonyl (Fmoc) solid phase peptide synthesis (SPPS). The vaccine constructs were designed with an alanine spacer between the O-linked mannose moieties to investigate the impact of distance between the mannose units on receptor-mediated uptake and/or binding in APCs. Uptake studies performed on F4/80(+) and CD11c(+) cells showed significant uptake and/or binding for lipopeptides containing mannose, and also the lipopeptide without mannose when compared to the control peptides (peptide with no lipid and peptide with no mannose and no lipid). Furthermore, mannan inhibition assays demonstrated that uptake of the mannosylated and lipidated peptides was receptor mediated. To address the specificity of receptor uptake, surface plasmon resonance studies were performed using biacore technology and confirmed high affinity of the mannosylated and lipidated vaccine constructs toward the MR. These studies confirm that both mannose and lipid moieties play significant roles in receptor-mediated uptake on APCs, potentially facilitating vaccine development.
authors.forename: Bita
authors.initials: B
authors.lastname: Sedaghat
authors.forename: Rachel J
authors.initials: RJ
authors.lastname: Stephenson
authors.forename: Ashwini Kumar
authors.initials: AK
authors.lastname: Giddam
authors.forename: Sharareh
authors.initials: S
authors.lastname: Eskandari
authors.affiliation: Infectious Diseases Program, QIMR Berghofer Medical Research Institute , Brisbane, Queensland 4029, Australia.
authors.forename: Simon H
authors.initials: SH
authors.lastname: Apte
authors.affiliation: Infectious Diseases Program, QIMR Berghofer Medical Research Institute , Brisbane, Queensland 4029, Australia.
authors.forename: David J
authors.initials: DJ
authors.lastname: Pattinson
authors.affiliation: Infectious Diseases Program, QIMR Berghofer Medical Research Institute , Brisbane, Queensland 4029, Australia.
authors.forename: Denise L
authors.initials: DL
authors.lastname: Doolan
authors.affiliation: School of Pharmacy, The University of Queensland , Woolloongabba, Queensland 4012, Australia.
authors.forename: Istvan
authors.initials: I
authors.lastname: Toth
mesh_terms: D000595:Amino Acid Sequence; D000938:Antigen-Presenting Cells / Q000378:metabolism; D037181:Lectins, C-Type / Q000378:metabolism*; D055666:Lipopeptides / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D008358:Mannose / Q000737:chemistry*; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000378:metabolism*
chemical_list: D037181:Lectins, C-Type; D055666:Lipopeptides; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins; D011956:Receptors, Cell Surface; D008358:Mannose