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1.
ACS Appl Bio Mater ; 7(6): 3877-3889, 2024 Jun 17.
Article in English | MEDLINE | ID: mdl-38832760

ABSTRACT

Adjuvants and immunomodulators that effectively drive a Th17-skewed immune response are not part of the standard vaccine toolkit. Vaccine adjuvants and delivery technologies that can induce Th17 or Th1/17 immunity and protection against bacterial pathogens, such as tuberculosis (TB), are urgently needed. Th17-polarized immune response can be induced using agonists that bind and activate C-type lectin receptors (CLRs) such as macrophage inducible C-type lectin (Mincle). A simple but effective strategy was developed for codelivering Mincle agonists with the recombinant Mycobacterium tuberculosis fusion antigen, M72, using tunable silica nanoparticles (SNP). Anionic bare SNP, hydrophobic phenyl-functionalized SNP (P-SNP), and cationic amine-functionalized SNP (A-SNP) of different sizes were coated with three synthetic Mincle agonists, UM-1024, UM-1052, and UM-1098, and evaluated for adjuvant activity in vitro and in vivo. The antigen and adjuvant were coadsorbed onto SNP via electrostatic and hydrophobic interactions, facilitating multivalent display and delivery to antigen presenting cells. The cationic A-SNP showed the highest coloading efficiency for the antigen and adjuvant. In addition, the UM-1098-adsorbed A-SNP formulation demonstrated slow-release kinetics in vitro, excellent stability over 12 months of storage, and strong IL-6 induction from human peripheral blood mononuclear cells. Co-adsorption of UM-1098 and M72 on A-SNP significantly improved antigen-specific humoral and Th17-polarized immune responses in vivo in BALB/c mice relative to the controls. Taken together, A-SNP is a promising platform for codelivery and proper presentation of adjuvants and antigens and provides the basis for their further development as a vaccine delivery platform for immunization against TB or other diseases for which Th17 immunity contributes to protection.


Subject(s)
Antigens, Bacterial , Lectins, C-Type , Nanoparticles , Silicon Dioxide , Th17 Cells , Lectins, C-Type/metabolism , Lectins, C-Type/immunology , Lectins, C-Type/agonists , Nanoparticles/chemistry , Th17 Cells/immunology , Animals , Silicon Dioxide/chemistry , Mice , Antigens, Bacterial/immunology , Antigens, Bacterial/administration & dosage , Antigens, Bacterial/chemistry , Mycobacterium tuberculosis/immunology , Adjuvants, Immunologic/chemistry , Adjuvants, Immunologic/pharmacology , Adjuvants, Immunologic/administration & dosage , Biocompatible Materials/chemistry , Biocompatible Materials/pharmacology , Particle Size , Materials Testing , Humans , Female , Membrane Proteins/immunology , Membrane Proteins/agonists
2.
J Med Chem ; 66(20): 13900-13917, 2023 10 26.
Article in English | MEDLINE | ID: mdl-37847244

ABSTRACT

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.


Subject(s)
Adjuvants, Vaccine , Toll-Like Receptor 4 , Animals , Mice , Immunologic Factors , Adjuvants, Immunologic/pharmacology , Adjuvants, Immunologic/chemistry , Ligands , Antibodies, Viral
3.
Clin Transl Sci ; 14(4): 1292-1302, 2021 07.
Article in English | MEDLINE | ID: mdl-33503331

ABSTRACT

The frequencies of genetic variants in the CYP3A4 and CYP3A5 genes differ greatly across global populations, leading to profound differences in the metabolic activity of these enzymes and resulting drug metabolism rates, with important consequences for therapeutic safety and efficacy. Yet, the impact of genetic variants on enzyme activity are incompletely described, particularly in American Indian and Alaska Native (AIAN) populations. To characterize genetic variation in CYP3A4 and CYP3A5 and its effect on enzyme activity, we partnered with AIAN people living in two regions of Alaska: Yup'ik Alaska Native people living in the Yukon-Kuskokwim Delta region of rural southwest Alaska and AIAN people receiving care at the Southcentral Foundation in Anchorage, Alaska. We identified low frequencies of novel and known variation in CYP3A4 and CYP3A5, including low frequencies of the CYP3A4*1G and CYP3A5*1 variants, and linkage disequilibrium patterns that differed from those we previously identified in an American Indian population in western Montana. We also identified increased activity of the CYP3A4*1G allele in vitro and in vivo. We demonstrated that the CYP3A4*1G allele confers increased protein content in human lymphoblastoid cells and both increased protein content and increased activity in human liver microsomes. We confirmed enhanced CYP3A4-mediated 4ß-vitamin D hydroxylation activity in Yup'ik people with the CYP3A4*1G allele. AIAN people in Alaska and Montana who carry the CYP3A4*1G allele-coupled with low frequency of the functional CYP3A5*1 variant-may metabolize CYP3A substrates more rapidly than people with the reference CYP3A4 allele.


Subject(s)
/genetics , Cytochrome P-450 CYP3A/metabolism , Indians, North American/genetics , Xenobiotics/metabolism , Adolescent , Adult , Aged , Aged, 80 and over , Alleles , Cell Line, Tumor , Child , Child, Preschool , Cytochrome P-450 CYP3A/genetics , Enzyme Assays , Female , Humans , Infant , Infant, Newborn , Linkage Disequilibrium , Male , Microsomes, Liver , Middle Aged , Pharmacogenomic Testing , Pharmacogenomic Variants , Polymorphism, Single Nucleotide , Young Adult
4.
Viruses ; 12(3)2020 03 03.
Article in English | MEDLINE | ID: mdl-32138181

ABSTRACT

Zika virus (ZIKV) is an emergent member of the Flaviviridae family which causes severe congenital defects and other major sequelae, but the cellular processes that support ZIKV replication are incompletely understood. Related flaviviruses use the endoplasmic reticulum (ER) as a membranous platform for viral replication and induce ER stress during infection. Our data suggest that ZIKV activates IRE1α, a component of the cellular response to ER stress. IRE1α is an ER-resident transmembrane protein that possesses a cytosolic RNase domain. Upon activation, IRE1α initiates nonconventional cytoplasmic splicing of XBP1 mRNA. Spliced XBP1 encodes a transcription factor, which upregulates ER-related targets. We find that ZIKV infection induces XBP1 mRNA splicing and induction of XBP1 target genes. Small molecule inhibitors of IRE1α, including those specific for the nuclease function, prevent ZIKV-induced cytotoxicity, as does genetic disruption of IRE1α. Optimal ZIKV RNA replication requires both IRE1α and XBP1. Spliced XBP1 has been described to cause ER expansion and remodeling and we find that ER redistribution during ZIKV infection requires IRE1α nuclease activity. Finally, we demonstrate that inducible genetic disruption of IRE1α and XBP1 impairs ZIKV replication in a mouse model of infection. Together, our data indicate that the ER stress response component IRE1α promotes ZIKV infection via XBP1 and may represent a potential therapeutic target.


Subject(s)
Endoribonucleases/metabolism , Host-Pathogen Interactions , Protein Serine-Threonine Kinases/metabolism , X-Box Binding Protein 1/metabolism , Zika Virus Infection/metabolism , Zika Virus Infection/virology , Zika Virus/physiology , Animals , Cells, Cultured , Chlorocebus aethiops , Endoplasmic Reticulum/metabolism , Endoplasmic Reticulum Stress , HeLa Cells , Humans , Protein Binding , Signal Transduction , Vero Cells , Virus Replication , Zika Virus Infection/pathology
5.
N Engl J Med ; 380(12): 1150-1157, 2019 03 21.
Article in English | MEDLINE | ID: mdl-30893535

ABSTRACT

A 58-year-old woman with debilitating ankylosing spondylitis who was born to consanguineous parents was found to have an apparent severe vitamin D deficiency that did not respond to supplementation. Liquid chromatography-tandem mass spectrometry showed the absence of circulating vitamin D-binding protein, and chromosomal microarray confirmed a homozygous deletion of the group-specific component (GC) gene that encodes the protein. Congenital absence of vitamin D-binding protein resulted in normocalcemia and a relatively mild disruption of bone metabolism, in this case complicated by severe autoimmune disease. (Funded by the National Institutes of Health and the University of Washington.).


Subject(s)
Autoimmune Diseases/complications , Gene Deletion , Hydroxycholecalciferols/blood , Spondylitis, Ankylosing/genetics , Vitamin D Deficiency/genetics , Vitamin D-Binding Protein/genetics , Calcium/blood , Chromatography, Liquid , Female , Fractures, Spontaneous/etiology , Gene Expression , Homozygote , Humans , Male , Middle Aged , Oligonucleotide Array Sequence Analysis , Polymorphism, Single Nucleotide , Siblings , Spondylitis, Ankylosing/complications , Tandem Mass Spectrometry , Vitamin D/metabolism , Vitamin D-Binding Protein/deficiency
6.
Sci Rep ; 8(1): 8989, 2018 06 12.
Article in English | MEDLINE | ID: mdl-29895962

ABSTRACT

There are many emerging and re-emerging globally prevalent viruses for which there are no licensed vaccines or antiviral medicines. Arbidol (ARB, umifenovir), used clinically for decades in several countries as an anti-influenza virus drug, inhibits many other viruses. In the current study, we show that ARB inhibits six different isolates of Zika virus (ZIKV), including African and Asian lineage viruses in multiple cell lines and primary human vaginal and cervical epithelial cells. ARB protects against ZIKV-induced cytopathic effects. Time of addition studies indicate that ARB is most effective at suppressing ZIKV when added to cells prior to infection. Moreover, ARB inhibits pseudoviruses expressing the ZIKV Envelope glycoprotein. Thus, ARB, a broadly acting anti-viral agent with a well-established safety profile, inhibits ZIKV, likely by blocking viral entry.


Subject(s)
Antiviral Agents/pharmacology , Indoles/pharmacology , Zika Virus Infection , Zika Virus/metabolism , A549 Cells , Animals , Chlorocebus aethiops , Cytopathogenic Effect, Viral/drug effects , HEK293 Cells , Humans , Vero Cells , Viral Envelope Proteins/metabolism , Zika Virus Infection/drug therapy , Zika Virus Infection/pathology
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