Do peptide mimetics of gp41 improve antibody-epitope interactions? Abstract In this F32 application I postulate that the broadly HIV neutralizing monoclonal antibodies, 2F5 and 4E10, against the membrane proximal external region (MPR) of gp41 were induced in a small population of donors by MPR epitopes due to modifications in the envelope proteins. My idea is to generate MPR immunogens incorporating unnatural amino acids that may induce stronger immune responses. I believe that these novel peptides will induce stronger affinity toward monoclonal antibodies, 2F5 and 4E10, and potentially induce higher titer polyclonal antibodies in a prime-boost immunization assay. The idea to use unnatural amino acids is unique. MPR immunogen design has focused on eliciting antibody responses that cross-react with phospholipids because the mAbs 2F5 and 4E10 exhibit unusual cross-reactivity with phospholipids. Incorporation of chemically modified amino acids into known epitopes may induce stronger immune responses. I will synthesize a group of MPR-based peptide sequences containing modified residues, which will be used to determine antibody affinity using a BIAcore assay. Structure activity relationships will be derived from these data and will guide subsequent rounds of chemically modified peptides to find sequences that will bind optimally to the monoclonal antibodies. I will then determine the immunogenicity of each peptide in rabbits using a prime-boost immunization strategy. Lipopeptides based on the peptides used in the BIAcore assay will be presented in a liposomal formulation to further enhance immunogenicity by presenting the epitopes in a lipid bilayer. Rabbits will be immunized subcutaneously and intramuscularly and antisera from these animals will be obtained and evaluated in a validated ELISA assay to determine the antibody titer induced by each peptide immunogen. Furthermore, formulations containing the same lipopeptides will be delivered intranasally to evaluate the mucosal immune response. Formulations will be prepared in the presence and absence of a peptide that binds and is transported by the neonatal Fc receptor (FcRn). ELISA assays will be used to determine the antibody responses in serum from SC/IM immunized animals, while mucosal lavage and serum samples will be evaluated from IN immunized animals. Differences in antisera titers from SC/IM immunized animals will be compared to IN immunized animals. Antisera and mucosal lavage samples with high titers in rabbits will be used in traditional inhibition assays to determine the neutralization activity of primary HIV-1 isolates in PBMCs. Success in generating neutralizing antibodies in rabbits will lead to future work to generate monoclonal antibodies. The chemically modified peptides that induce neutralizing antibodies in rabbits could provide a widely applicable epitope design strategy for the development of vaccines to a variety of infectious diseases including HIV.
The development of a vaccine against HIV has not yet been developed due to the high mutation rate of the virus. Unsuccessful vaccine development thus far may be due to the use of improper immunogens, which may be addressed using unnatural amino acids. This proposal focuses on the development of novel peptide mimetics as potential HIV vaccines with the potential to generate broadly neutralizing antibodies.
|Kierstead, Paul H; Okochi, Hideaki; Venditto, Vincent J et al. (2015) The effect of polymer backbone chemistry on the induction of the accelerated blood clearance in polymer modified liposomes. J Control Release 213:1-9|
|Venditto, Vincent J; Dolor, Aaron; Kohli, Aditya et al. (2014) Sulfated quaternary amine lipids: a new class of inverse charge zwitterlipids. Chem Commun (Camb) 50:9109-11|
|Kohli, Aditya G; Kierstead, Paul H; Venditto, Vincent J et al. (2014) Designer lipids for drug delivery: from heads to tails. J Control Release 190:274-87|
|Venditto, Vincent J; Wieczorek, Lindsay; Molnar, Sebastian et al. (2014) Chemically modified peptides based on the membrane-proximal external region of the HIV-1 envelope induce high-titer, epitope-specific nonneutralizing antibodies in rabbits. Clin Vaccine Immunol 21:1086-93|
|Julien, Olivier; Kampmann, Martin; Bassik, Michael C et al. (2014) Unraveling the mechanism of cell death induced by chemical fibrils. Nat Chem Biol 10:969-76|
|Venditto, Vincent J; Watson, Douglas S; Motion, Michael et al. (2013) Rational design of membrane proximal external region lipopeptides containing chemical modifications for HIV-1 vaccination. Clin Vaccine Immunol 20:39-45|
|Venditto, Vincent J; Szoka Jr, Francis C (2013) Cancer nanomedicines: so many papers and so few drugs! Adv Drug Deliv Rev 65:80-8|
|Watson, Douglas S; Platt, Virginia M; Cao, Limin et al. (2011) Antibody response to polyhistidine-tagged peptide and protein antigens attached to liposomes via lipid-linked nitrilotriacetic acid in mice. Clin Vaccine Immunol 18:289-97|