Presentation of antigen on extracellular vesicles using transmembrane domains from viral glycoproteins for enhanced immunogenicity

A vaccine antigen, when launched as DNA or RNA, can be presented in various forms, including intracellular, secreted, membrane-bound, or on extracellular vesicles (EVs). Whether an antigen in one or more of these forms is superior in immune induction remains unclear. In this study, we used GFP as a model antigen and first compared the EV-loading efficiency of transmembrane domains (TMs) from various viral glycoproteins, and then investigated whether EV-bound GFP (EV-GFP) would enhance immune induction. Our data showed that GFP fused to viral TMs was successfully loaded onto the surface of EVs. In addition, GFP-bound EVs were predominantly associated with the exosome marker CD81. Immunogenicity study with EV-GFP-producing plasmids in mice demonstrated that antigen-specific IgG and IgA were significantly increased in EV-GFP groups, compared to soluble and intracellular GFP groups. Similarly, GFP-specific T cell response-related cytokines produced by antigen-stimulated splenocytes were also enhanced in mice immunized with EV-GFP constructs. Immunogenicity study with purified soluble GFP and GFP EVs further confirmed the immune enhancement property of EV-GFP in mice. In vitro uptake assays indicated that EV-GFP was more efficiently taken up than soluble GFP by mouse splenocytes and such uptake was B cell preferential. Taken together, our data indicate that viral TMs can efficiently load antigens onto the EV surface, and that EV-bound antigen enhances both humoral and cell-mediated antigen-specific responses

The role of zinc in the biocorrosion behavior of resorbable Mg‒Zn‒Ca alloys

ISSN:1742-7061ISSN:1878-756

Precipitation in lean Mg-Zn-Ca alloys

While lean Mg-Zn-Ca alloys are promising materials for temporary implants, questions remain on the impact of Zn and Ca on the microstructure. In this context, the precipitation of Zn and Ca in Mg-1.5Zn-0.25Ca (in wt.%), initially extruded at 330 & DEG;C, towards Mg-Ca binary precipitates or Ca-Mg-Zn ternary precipitates was probed in a multiscale correlative approach using atom probe tomography (APT) and an-alytical transmission electron microscopy (TEM). Particular focus was set on the ternary precipitate phase whose structure is debated. In the as-extruded material, the binary precipitates are made of hexagonal C14 Mg2Ca containing up to about 3 at.% of Zn. The ternary ones are based on the hexagonal Ca2Mg5Zn5 prototype structure with a composition close to Ca3Mg11Zn4, as deduced from atomically resolved EDS mapping and scanning TEM imaging, supported by simulations. The precipitation sequence was scru-tinized upon linear heating from room temperature to 375 ?, starting from the solutionized material. Three exothermic differential scanning calorimetry (DSC) peaks were observed, at respectively 125, 250 and 320 & DEG;C. Samples were taken after the peak decays, at respectively 205, 260 and 375? for structural analysis. At 205 & DEG;C, APT analysis revealed Ca-rich, Zn-rich and Zn-Ca-rich clusters of about 3 nm in size and with a number density of 5.7 x 10 23 m -3. At 260 ?, APT and TEM showed mono-layered Zn-Ca-rich Guinier-Preston (GP) zones of about 8 nm in size and with a number density of 1.3 x 10 23 m -3. At 375 ?, larger and highly coherent elongated precipitates were found, with a size of about 50 nm. They occur as binary Mg-Ca precipitates or ternary Ca2Mg6Zn3 precipitates, as deduced from scanning TEM-based energy dispersive X-ray spectroscopy (EDS) and nanodiffraction in TEM. Here, the binary precipitates outnumber the ternary ones, while in the as-extruded material the ternary precipitates outnumber the binary ones, which corresponds well to the calculated phase diagram. We correlated the microstruc-ture to hardness probed by Vickers testing. The largest hardening relates to the end of the 125 ? DSC peak and thus to GP zones, which outperform the hardening induced by the nanometer-sized clusters and the larger intermetallic particles. The complexity of the precipitation sequence in lean Mg-Zn-Ca alloys is discussed.(c) 2022 The Authors. Published by Elsevier Ltd on behalf of Acta Materialia Inc. This is an open access article under the CC BY license ( http://creativecommons.org/licenses/by/4.0/

Investigation of the thermogelation of a promising biocompatible ABC triblock terpolymer and its comparison with pluronic F127

Thermoresponsive polymers with the appropriate structure form physical networks upon changes in temperature, and they find utility in formulation science, tissue engineering, and drug delivery. Here, we report a cost-effective biocompatible alternative, namely OEGMA30015-b-BuMA26-b-DEGMA13, which forms gels at low concentrations (as low as 2% w/w); OEGMA300, BuMA, and DEGMA stand for oligo(ethylene glycol) methyl ether methacrylate (MM = 300 g mol–1), n-butyl methacrylate, and di(ethylene glycol) methyl ether methacrylate, respectively. This polymer is investigated in depth and is compared to its commercially available counterpart, Poloxamer P407 (Pluronic F127). To elucidate the differences in their macroscale gelling behavior, we investigate their nanoscale self-assembly by means of small-angle neutron scattering and simultaneously recording their rheological properties. Two different gelation mechanisms are revealed. The triblock copolymer inherently forms elongated micelles, whose length increases by temperature to form worm-like micelles, thus promoting gelation. In contrast, Pluronic F127’s micellization is temperature-driven, and its gelation is attributed to the close packing of the micelles. The gel structure is analyzed through cryogenic scanning and transmission electron microscopy. Ex vivo gelation study upon intracameral injections demonstrates excellent potential for its application to improve drug residence in the eye

Mitigating the detrimental effects of galvanic corrosion by nanoscale composite architecture design

Widespread application of magnesium (Mg) has been prevented by its low strength and poor corrosion resistance. Core of this limitation is Mg's low electrochemical potential and low solubility for most elements, favoring secondary phase precipitation acting as effective micro-galvanic elements. Mg-based metal-metal composites, while benefiting strength, are similarly active galvanic couples. We show that related detrimental corrosion susceptibility is overcome by nanoscale composite architecture design. Nanoscale phase spacings enable high-strength Mg-Fe composites with degradation rates as low as ultra-high purity Mg. Our concept thus fundamentally changes today's understanding of Mg's corrosion and significantly widens the property space of Mg-based materials.ISSN:2397-210

Investigation of the thermogelation of a promising biocompatible ABC triblock terpolymer and its comparison with pluronic F127

Thermoresponsive polymers with the appropriate structure form physical networks upon changes in temperature, and they find utility in formulation science, tissue engineering, and drug delivery. Here, we report a cost-effective biocompatible alternative, namely OEGMA30015-b-BuMA26-b-DEGMA13, which forms gels at low concentrations (as low as 2% w/w); OEGMA300, BuMA, and DEGMA stand for oligo(ethylene glycol) methyl ether methacrylate (MM = 300 g mol–1), n-butyl methacrylate, and di(ethylene glycol) methyl ether methacrylate, respectively. This polymer is investigated in depth and is compared to its commercially available counterpart, Poloxamer P407 (Pluronic F127). To elucidate the differences in their macroscale gelling behavior, we investigate their nanoscale self-assembly by means of small-angle neutron scattering and simultaneously recording their rheological properties. Two different gelation mechanisms are revealed. The triblock copolymer inherently forms elongated micelles, whose length increases by temperature to form worm-like micelles, thus promoting gelation. In contrast, Pluronic F127’s micellization is temperature-driven, and its gelation is attributed to the close packing of the micelles. The gel structure is analyzed through cryogenic scanning and transmission electron microscopy. Ex vivo gelation study upon intracameral injections demonstrates excellent potential for its application to improve drug residence in the eye