Immunogenicity Of Peptide Vaccine Research Articles

Expression and purification of Brucella spp lumazine synthase decameric carrier in fusion to extracellular domain of influenza M2E protein

Brucella spp. Lumazine synthase enzyme, is a decameric protein carrier that display foreign antigens effectively in a polyvalent manner. The applied strategy using with this molecule results in a higher density of antigen and enhancing immunogenicity of peptide vaccines. In the current study, Brucella lumazine synthase (BLS) was applied for fusion with influenza matrix protein 2 ecto domain (M2E) as a foreign peptide. The primary studies were based on bioinformatics tools and the fusion was expressed and purified in following levels. Forming of decamer was confirmed by electrophoresis and western blotting techniques. Influenza matrix protein 2 was stably expressed at the 10 amino terminals of lumazine synthase. The purified fusion was injected to mice and immune responses were evaluated with indirect enzyme linked immunosorbent assay (ELISA) technique. According to ELISA results yield of purification process was 41% with ion exchange method and the protein was as a single band in Sodium Dodecyl Sulfate PolyAcrylamide Gel Electrophoresis (SDS-PAGE). The titer of immunized mice serum with decameric fusion of lumazine synthase and matrix protein (M2BL) was determined to be more than 1:32000 by indirect ELISA. The level of responses against matrix protein in decameric state of M2BL, was about 20% higher than monomer M2BL. Anti M2BL was cross reacted effectively with influenza M2E and in comparison with samples injected with adjuvant the level of antiM2E was similar. The results in this study confirms the role of multi copy presentation systems and applicability of BLS as an antigen carrier and adjuvant in designing peptide vaccines.

Enhancement of Peptide Vaccine Immunogenicity by Increasing Lymphatic Drainage and Boosting Serum Stability

Antitumor T-cell responses have the potential to be curative in cancer patients, but the induction of potent T-cell immunity through vaccination remains a largely unmet goal of immunotherapy. We previously reported that the immunogenicity of peptide vaccines could be increased by maximizing delivery to lymph nodes (LNs), where T-cell responses are generated. This was achieved by conjugating the peptide to 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-PEG (DSPE-PEG) to promote albumin binding, which resulted in enhanced lymphatic drainage and improved T-cell responses. Here, we expanded upon these findings and mechanistically dissected the properties that contribute to the potency of this amphiphile-vaccine (amph-vaccine). We found that multiple linkage chemistries could be used to link peptides with DSPE-PEG, and further, that multiple albumin-binding moieties conjugated to peptide antigens enhanced LN accumulation and subsequent T-cell priming. In addition to enhancing lymphatic trafficking, DSPE-PEG conjugation increased the stability of peptides in serum. DSPE-PEG peptides trafficked beyond immediate draining LNs to reach distal nodes, with antigen presented for at least a week in vivo, whereas soluble peptide presentation quickly decayed. Responses to amph-vaccines were not altered in mice deficient in the albumin-binding neonatal Fc receptor (FcRn), but required Batf3-dependent dendritic cells (DCs). Amph-peptides were processed by human DCs equivalently to unmodified peptides. These data define design criteria for enhancing the immunogenicity of molecular vaccines to guide the design of next-generation peptide vaccines. Cancer Immunol Res; 6(9); 1025-38. ©2018 AACR.