Highlights • Seven TLR7/8 agonists were synthesized and analyzed for their adjuvanting capacity. • Agonists stimulated cytokine production and matured both mDCs and pDCs. • The oxoadenine compounds ...were more potent than the imidazoquinolines. • An oxoadenine compound augmented the adaptive immune response to antigen in pigs.
Most licensed seasonal influenza vaccines are non-adjuvanted and rely primarily on vaccine-induced antibody titers for protection. As such, seasonal antigenic drift and suboptimal vaccine strain ...selection often results in reduced vaccine efficacy. Further, seasonal H3N2 influenza vaccines demonstrate poor efficacy compared to H1N1 and influenza type B vaccines. New vaccines, adjuvants, or delivery technologies that can induce broader or cross-seasonal protection against drifted influenza virus strains, likely through induction of protective T cell responses, are urgently needed. Here, we report novel lipidated TLR7/8 ligands that act as strong adjuvants to promote influenza-virus specific Th1-and Th17-polarized T cell responses and humoral responses in mice with no observable toxicity. Further, the adjuvanted influenza vaccine provided protection against a heterologous H3N2 influenza challenge in mice. These responses were further enhanced when combined with a synthetic TLR4 ligand adjuvant. Despite differences between human and mouse TLR7/8, these novel lipidated imidazoquinolines induced the production of cytokines required to polarize a Th1 and Th17 immune response in human PBMCs providing additional support for further development of these compounds as novel adjuvants for the induction of broad supra-seasonal protection from influenza virus.
Abstract
Infection is the most common cause of mortality early in life, yet the broad potential of immunization is not fully realized in this vulnerable population. Most vaccines are administered ...during infancy and childhood, but in some cases the full benefit of vaccination is not realized in-part. New adjuvants are cardinal to further optimize current immunization approaches for early life. However, only a few classes of adjuvants are presently incorporated in vaccines approved for human use. Recent advances in the discovery and delivery of Toll-like receptor (TLR) agonist adjuvants have provided a new toolbox for vaccinologists. Prominent among these candidate adjuvants are synthetic small molecule TLR7/8 agonists. The development of an effective infant
Bordetella pertussis
vaccine is urgently required because of the resurgence of pertussis in many countries, contemporaneous to the switch from whole cell to acellular vaccines. In this context, TLR7/8 adjuvant based vaccine formulation strategies may be a promising tool to enhance and accelerate early life immunity by acellular
B. pertussis
vaccines. In the present study, we optimized (a) the formulation delivery system, (b) structure, and (c) immunologic activity of novel small molecule imidazoquinoline TLR7/8 adjuvants towards human infant leukocytes, including dendritic cells. Upon immunization of neonatal mice, this TLR7/8 adjuvant overcame neonatal hyporesponsiveness to acellular pertussis vaccination by driving a T helper (Th)1/Th17 biased T cell- and IgG2c-skewed humoral response to a licensed acellular vaccine (DTaP). This potent immunization strategy may represent a new paradigm for effective immunization against pertussis and other pathogens in early life.
Most known synthetic toll-like receptor 4 (TLR4) agonists are carbohydrate-based lipid-A mimetics containing several fatty acyl chains, including a labile 3-O-acyl chain linked to the C-3 position of ...the non-reducing sugar known to undergo cleavage impacting stability and resulting in loss of activity. To overcome this inherent instability, we rationally designed a new class of chemically more stable synthetic TLR4 ligands that elicit robust innate and adaptive immune responses. This new class utilized a diamino allose phosphate (DAP) scaffold containing a nonhydrolyzable 3-amide bond instead of the classical 3-ester. Accordingly, the DAPs have significantly improved thermostability in aqueous formulations and potency relative to other known natural and synthetic TLR4 ligands. Furthermore, the DAP analogues function as potent vaccine adjuvants to enhance influenza-specific antibodies in mice and provide protection against lethal influenza virus challenges. This novel set of TLR4 ligands show promise as next-generation vaccine adjuvants and stand-alone immunomodulators.
Despite the ease of production and improved safety profiles of recombinant vaccines, the inherently low immunogenicity of unadjuvanted proteins remains an impediment to their widespread adoption. The ...covalent tethering of TLR agonists to antigenic proteins offers a unique approach to co-deliver both constituents to the same cell-enhancing vaccine efficacy while minimizing reactogenicity. However, the paucity of simple and effective linker chemistries continues to hamper progress. Here, we present a modular, PEG-based linker system compatible with even extremely lipophilic and challenging TLR7/8 agonists. To advance the field and address previous obstacles, we offer the most straightforward and antigen-preserving linker system to date. These antigen-adjuvant conjugates enhance antigen-specific immune responses in mice, demonstrating the power of our approach within the context of modern vaccinology.
Co-delivery of antigens and adjuvants to the same antigen-presenting cells (APCs) can significantly improve the efficacy and safety profiles of vaccines. Here, we report amine-grafted silica ...nanoparticles (A-SNP) as a tunable vaccine co-delivery platform for TLR7/8 agonists along with the recombinant influenza antigen hemagglutinin H7 (H7) to APCs. A-SNP of two different sizes (50 and 200 nm) were prepared and coated with INI-4001 at different coating densities, followed by co-adsorption of H7. Both INI-4001 and H7 showed >90% adsorption to the tested A-SNP formulations. TNF-α and IFN-α cytokine release by human peripheral blood mononuclear cells as well as TNF-α, IL-6, and IL-12 release by mouse bone marrow-derived dendritic cells revealed that the potency of the INI-4001-adsorbed A-SNP (INI-4001/A-SNP) formulations was improved relative to aqueous formulation control. This improved potency was dependent on particle size and ligand coating density. In addition, slow-release profiles of INI-4001 were measured from INI-4001/A-SNP formulations in plasma with 30-50% INI-4001 released after 7 days. In vivo murine immunization studies demonstrated significantly improved H7-specific humoral and Th1/Th17-polarized T cell immune responses with no observed adverse reactions. Low-density 50 nm INI-4001/A-SNP elicited significantly higher IFN-γ and IL-17 induction over that of the H7 antigen-only group and INI-4001 aqueous formulation controls. In summary, this work introduces an effective and biocompatible SNP-based co-delivery platform that enhances the immunogenicity of TLR7/8 agonist-adjuvanted subunit influenza vaccines.
We present optical spectroscopy of galaxies in clusters detected through the Sunyaev-Zel'dovich (SZ) effect with the South Pole Telescope (SPT). We report our own measurements of 61 spectroscopic ...cluster redshifts, and 48 velocity dispersions each calculated with more than 15 member galaxies. This catalog also includes 19 dispersions of SPT-observed clusters previously reported in the literature. The majority of the clusters in this paper are SPT-discovered; of these, most have been previously reported in other SPT cluster catalogs, and five are reported here as SPT discoveries for the first time. By performing a resampling analysis of galaxy velocities, we find that unbiased velocity dispersions can be obtained from a relatively small number of member galaxies (lap30), but with increased systematic scatter. We use this analysis to determine statistical confidence intervals that include the effect of membership selection.We fit scaling relations between the observed cluster velocity dispersions and mass estimates from SZ and X-ray observables. In both cases, the results are consistent with the scaling relation between velocity dispersion and mass expected from dark-matter simulations. We measure a ~30% log-normal scatter in dispersion at fixed mass, and a ~10% offset in the normalization of the dispersion-mass relation when compared to the expectation from simulations, which is within the expected level of systematic uncertainty.
We present two luminous UV/optical flares from the nuclei of apparently inactive early-type galaxies at image and 0.33 that have the radiative properties of a flare from the tidal disruption of a ...star. In this paper we report the second candidate tidal disruption event discovery in the UV by the GALEX Deep Imaging Survey and present simultaneous optical light curves from the CFHTLS Deep Imaging Survey for both UV flares. The first few months of the UV/optical light curves are well fitted with the canonical image power-law decay predicted for emission from the fallback of debris from a tidally disrupted star. Chandra ACIS X-ray observations during the flares detect soft X-ray sources with image or image and place limits on hard X- ray emission from an underlying AGN down to image. Blackbody fits to the UV/optical spectral energy distributions of the flares indicate peak flare luminosities of image10 super(44)-10 super(45) ergs s super(-1). The temperature, luminosity, and light curves of both flares are in excellent agreement with emission from a tidally disrupted main-sequence star onto a central black hole of several times 10 super(7) M sub(image). The observed detection rate of our search over image2.9 deg super( 2) of GALEX Deep Imaging Survey data spanning from 2003 to 2007 is consistent with tidal disruption rates calculated from dynamical models, and we use these models to make predictions for the detection rates of the next generation of optical synoptic surveys.
The Blanco Cosmology Survey (BCS) is a 60 night imaging survey of ~80 deg super(2) of the southern sky located in two fields: (alpha, delta) = (5 hr, -55degrees) and (23 hr, -55degrees). The survey ...was carried out between 2005 and 2008 in griz bands with the Mosaic2 imager on the Blanco 4 m telescope. The primary aim of the BCS survey is to provide the data required to optically confirm and measure photometric redshifts for Sunyaev-Zel'dovich effect selected galaxy clusters from the South Pole Telescope and the Atacama Cosmology Telescope. We process and calibrate the BCS data, carrying out point-spread function-corrected model-fitting photometry for all detected objects. The median 10sigma galaxy (point-source) depths over the survey in griz are approximately 23.3 (23.9), 23.4 (24.0), 23.0 (23.6), and 21.3 (22.1), respectively. The astrometric accuracy relative to the USNO-B survey is ~45 mas. We calibrate our absolute photometry using the stellar locus in grizJ bands, and thus our absolute photometric scale derives from the Two Micron All Sky Survey, which has ~2% accuracy. The scatter of stars about the stellar locus indicates a systematic floor in the relative stellar photometric scatter in griz that is ~1.9%, ~2.2%, ~2.7%, and ~2.7%, respectively. A simple cut in the AstrOmatic star-galaxy classifier spread_model produces a star sample with good spatial uniformity. We use the resulting photometric catalogs to calibrate photometric redshifts for the survey and demonstrate scatter deltaz/(1 + z) = 0.054 with an outlier fraction eta < 5% to z ~ 1. We highlight some selected science results to date and provide a full description of the released data products.
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