Combined enteropathogen recombinant construct
Inventors
Guerry, Patricia • Monteiro, Mario Artur • Savarino, Stephen
Assignees
University of Guelph • US Department of Navy
Publication Number
US-11077200-B2
Publication Date
2021-08-03
Expiration Date
2026-01-10
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Abstract
The inventive subject matter relates to a construct comprising antigens derived from multiple enterobacteria including Campylobacter jejuni capsule polysaccharide polymer, enterotoxigenic Escherichia coli recombinant polypeptide construct and lipopolysaccharide from Shigella spp. The subject invention also relates to a method of inducing an immune response utilizing the inventive composition.
Core Innovation
The invention relates to an immunogenic construct combining antigens derived from multiple enterobacteria, specifically comprising a polypeptide construct expressing enterotoxigenic Escherichia coli (ETEC) fimbrial subunits combined with a Campylobacter jejuni capsule polysaccharide or Shigella spp. lipopolysaccharide (LPS). The construct can include one or more Campylobacter jejuni capsule polysaccharides conjugated to one or more ETEC recombinant polypeptide constructs, or Shigella LPS conjugated to the ETEC polypeptide construct. The ETEC recombinant polypeptide construct includes major and/or minor fimbrial subunits connected via polypeptide linkers and stabilized by donor strand complementation, with features such as N-terminal deletions to avoid undesirable multimer associations.
The problem addressed is the significant global health threat posed by enteropathogenic bacteria, including ETEC, Shigella spp., and Campylobacter jejuni, which cause major bacterial diarrheal diseases worldwide, especially in young children and travelers. No FDA-licensed vaccines are currently available for these pathogens, and challenges such as the diversity of ETEC colonization factors and the risk of autoimmune complications like Guillain-Barré Syndrome from C. jejuni lipooligosaccharide have impeded vaccine development. The invention aims to provide a combined immunogenic composition capable of eliciting immune responses against multiple pathogens while overcoming these challenges.
Claims Coverage
The claims define multiple inventive features focused on a multi-agent immunogenic construct combining Campylobacter jejuni capsule polysaccharide with an Escherichia coli enterotoxigenic recombinant polypeptide construct and related methods for inducing immune responses.
Multi-agent immunogenic construct combining Campylobacter jejuni capsule polysaccharide with ETEC recombinant polypeptide construct
The construct comprises Campylobacter jejuni capsule polysaccharide conjugated to a protein carrier comprising an Escherichia coli enterotoxigenic recombinant polypeptide construct, wherein the ETEC polypeptide construct includes minor or major fimbrial subunits connected via polypeptide linkers. Each major fimbrial subunit contains a donor β strand and is connected to others by polypeptide linkers, with a C-terminal major subunit linked to a donor β strand derived from a homologous or heterologous major subunit. The construct can also include multiple recombinant polypeptide constructs comprising fimbrial subunits from different fimbrial types and optionally a C-terminal histidine tag.
Molar ratio and composition includes Shigella lipopolysaccharide
The construct features a molar ratio of Campylobacter jejuni polysaccharide to Escherichia coli recombinant protein carrier of about 1:1 to 5:1, and may include Shigella lipopolysaccharide in addition to the Campylobacter jejuni capsule polysaccharide, expanding antigenic coverage.
Specific structural components of the polysaccharide and polypeptide elements
The polysaccharide structures comprise repeating trisaccharides with defined formulas representing Campylobacter jejuni capsule types. The Shigella lipopolysaccharide component includes specific structural features exemplified by Shigella flexneri 2a LPS. The donor β strand of the ETEC fimbrial major subunits contains 12 to 16 amino acids, and minor or major subunits may retain an 18-22 amino acid signal peptide. Polypeptide linkers are sequences such as SEQ ID No. 5 or tri-glycine, and major subunits may include a deletion of 14 to 18 N-terminal amino acids.
Method of inducing immune responses using the multi-agent immunogenic construct
A method is claimed of inducing immune responses against Campylobacter jejuni, comprising administration of the multi-agent immunogenic composition at defined dose ranges, followed by one or more boosting doses. The method applies to compositions containing the described polysaccharide structures and optionally including Shigella lipopolysaccharide, and incorporates the structural features of the polypeptide constructs.
The claims cover a combined enteropathogen immunogenic construct that links Campylobacter jejuni capsule polysaccharides and optionally Shigella LPS to stabilized ETEC fimbrial recombinant polypeptide constructs. The structural features of protein subunits, linkers, donor β strands, and polysaccharide structures are detailed, supporting an invention directed to multi-pathogen vaccine constructs and methods of inducing immune responses.
Stated Advantages
The composition induces immune responses against multiple bacterial components simultaneously, e.g., ETEC fimbriae and Campylobacter jejuni capsule polysaccharides.
Use of capsule polysaccharide from C. jejuni reduces risk of inducing Guillain-Barré Syndrome compared to lipooligosaccharide-based vaccines.
Donor strand complementation stabilizes ETEC fimbrial subunits, enhancing immunogenicity and protease resistance.
The multipartite construct design enables broad immunogenic coverage across diverse ETEC fimbrial types and strains.
Conjugation serves as a protein carrier enhancing immunogenicity of polysaccharide antigens from Campylobacter jejuni and Shigella.
Documented Applications
Vaccination for prevention of bacterial diarrheal diseases caused by enteropathogenic Escherichia coli, Campylobacter jejuni, and Shigella spp.
Induction of immune responses in mammals, including humans, to multiple enterobacterial pathogens using a combined antigen composition.
Use of multipartite fusion protein constructs as carriers for bacterial polysaccharide antigens in vaccine formulations.
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