Design of lipid nanoparticle delivery agents for multivalent display of recombinant Env trimers in HIV vaccination

Background: Immunization strategies that elicit antibodies capable of neutralizing diverse strains of the virus will likely be an important part of a successful vaccine against HIV. The envelope trimer is the only neutralizing target on the virus, and strategies to promote durable, high avidity antibody responses against the native intact trimer structure are lacking. We recently developed chemically-crosslinked lipid nanocapsules as carriers of molecular adjuvants and encapsulated or surface-displayed antigens, which promote follicular helper T-cell responses and elicited high-avidity, durable antibody responses to a candidate malaria antigen (Moon et al. Nat. Mater. 10 243 (2011); Moon et al. PNAS 109 1080 (2012)). Methods: To apply this system to the delivery of HIV antigens, we developed a strategy to anchor recombinant envelope trimers to the surfaces of these particles under conditions preserving the antigenic integrity of the trimers, allowing multivalent display of these immunogens for immunization. To anchor trimers in their native orientation, gp140 trimers with terminal his-tags were anchored to the surface of lipid nanocapsules via Ni-NTA-functionalized lipids. Results: Owing to their significant size (409 kDa) and heavy glycosylation, we found that liquid-ordered and/or gel-phase lipid compositions were required to stably anchor trimers to particle membranes. Trimer-loaded nanocapsules carrying monophosphoryl lipid A elicited durable antibody responses with titers comparable to a Complete Freund’s Adjuvant (CFA)-like emulsion in mice, without the toxic inflammation associated with the latter adjuvant. Further, nanocapsules elicited strong helper T-cell responses associated with a steadily increasing avidity of trimer-binding antibody over 90 days, which was not replicated by other adjuvants. Conclusion: These results suggest that nanoparticles displaying HIV trimers in an oriented, multivalent presentation can promote key aspects of the humoral response against Env immunogens.National Institutes of Health (U.S.) (AI095109)Massachusetts Institute of Technology. Ragon Institute of MGH, MIT, and Harvar

Design of Lipid Nanocapsule Delivery Vehicles for Multivalent Display of Recombinant Env Trimers in HIV Vaccination

Immunization strategies that elicit antibodies capable of neutralizing diverse virus strains will likely be an important part of a successful vaccine against HIV. However, strategies to promote robust humoral responses against the native intact HIV envelope trimer structure are lacking. We recently developed chemically cross-linked lipid nanocapsules as carriers of molecular adjuvants and encapsulated or surface-displayed antigens, which promoted follicular helper T-cell responses and elicited high-avidity, durable antibody responses to a candidate malaria antigen. To apply this system to the delivery of HIV antigens, Env gp140 trimers with terminal his-tags (gp140T-his) were anchored to the surface of lipid nanocapsules via Ni-NTA-functionalized lipids. Initial experiments revealed that the large (409 kDa), heavily glycosylated trimers were capable of extracting fluid phase lipids from the membranes of nanocapsules. Thus, liquid-ordered and/or gel-phase lipid compositions were required to stably anchor trimers to the particle membranes. Trimer-loaded nanocapsules combined with the clinically relevant adjuvant monophosphoryl lipid A primed high-titer antibody responses in mice at antigen doses ranging from 5 μg to as low as 100 ng, whereas titers dropped more than 50-fold over the same dose range when soluble trimer was mixed with a strong oil-in-water adjuvant comparator. Nanocapsule immunization also broadened the number of distinct epitopes on the HIV trimer recognized by the antibody response. These results suggest that nanocapsules displaying HIV trimers in an oriented, multivalent presentation can promote key aspects of the humoral response against Env immunogens