Combined enteropathogen recombinant construct
Inventors
Guerry, Patricia • Monteiro, Mario Artur • Savarino, Stephen
Assignees
University of Guelph • US Department of Navy
Publication Number
US-10105448-B2
Publication Date
2018-10-23
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 a multi-agent immunogenic construct that combines antigens derived from multiple enterobacteria, specifically Campylobacter jejuni capsule polysaccharide polymer, enterotoxigenic Escherichia coli (ETEC) recombinant polypeptide constructs, and lipopolysaccharides from Shigella spp. This construct aims to induce an immune response simultaneously against these significant bacterial pathogens known to cause diarrhea worldwide.
The problem addressed is the lack of FDA-licensed vaccines available for major enteric pathogens including ETEC, Shigella spp., and Campylobacter jejuni. ETEC, for example, causes substantial morbidity and mortality, especially in infants, young children, and travelers. Vaccine development for ETEC is complicated due to the diversity of colonization factors, the presence of different enterotoxins, and the need for broad coverage. Similarly, Campylobacter jejuni is a serious pathogen with no licensed vaccine and is also associated with severe sequelae such as Guillain-Barré Syndrome, which complicates immunization strategies.
To address these challenges, the invention presents a recombinant immunogenic construct that links ETEC fimbrial subunits, stabilized by donor strand complementation and connected via polypeptide linkers, to Campylobacter jejuni capsule polysaccharide or Shigella lipopolysaccharide. This enables broad-spectrum immunogenic coverage against multiple fimbrial types of ETEC (including Class 5, CS3, and CS6), while avoiding potential adverse autoimmune responses linked to molecular mimicry with the lipooligosaccharide of Campylobacter jejuni by using its capsule polysaccharide. The ETEC recombinant polypeptide also acts as a protein carrier enhancing the immunogenicity of the polysaccharide antigen. Multipartite fusion constructs connect subunits from different fimbrial types, potentially conjugated to multiple bacterial polysaccharides, to induce comprehensive immunity against diverse bacterial strains.
Claims Coverage
The patent contains a single independent claim focused on a multi-agent immunogenic construct. It outlines the main inventive features related to the design of the recombinant polypeptide constructs and their conjugation to bacterial polysaccharides.
Multi-agent immunogenic construct composition
The construct comprises a Campylobacter jejuni capsule polysaccharide conjugated to an Escherichia coli enterotoxigenic recombinant polypeptide construct that includes minor or major fimbrial subunits connected to one or more major fimbrial subunits of the same fimbrial type via polypeptide linkers. The subunits themselves are connected by polypeptide linkers, and the C-terminal major subunit is stabilized by a donor β strand from a homologous or heterologous major subunit containing 12 to 16 amino acids from the N-terminal region.
Multipartite recombinant polypeptide constructs
The recombinant polypeptide construct can be connected to one or more additional recombinant polypeptide constructs, where each contains fimbrial subunits derived from different fimbrial types than others, allowing for broad coverage. The constructs may contain a C-terminal histidine tag for purification.
Replacement or addition of polysaccharides
Shigella lipopolysaccharide (LPS) can be used in place of or combined with the Campylobacter jejuni capsule polysaccharide in the immunogenic construct, enabling multi-pathogen immunogenicity.
Specific molar ratio of polysaccharide to protein carrier
The molar ratio of Campylobacter jejuni or Shigella LPS to Escherichia coli recombinant protein carrier is specified within a range of 1:1 to 5:1 to optimize immunogenicity.
Sequence-specific embodiments
The Escherichia coli recombinant polypeptide construct includes specific amino acid and nucleotide sequences (e.g., SEQ ID Nos. 105, 108, 109, 110) defining the fimbrial subunits.
Polysaccharide structural features
Specific repeating trisaccharide structures of the Campylobacter jejuni capsule polysaccharide are identified for use in the construct.
Signal peptide and linker specifications
The minor or major subunits may contain an 18-22 amino acid N-terminal signal peptide. The polypeptide linkers connecting subunits are specified as either the sequence of SEQ ID No. 5 or tri-glycine linkers.
N-terminal deletions in subunits
One or more major subunits can incorporate deletions of 14 to 18 amino acids from their N-terminal region to prevent undesirable associations or oligomer formation.
The claims define a multi-agent immunogenic construct comprising recombinant E. coli fimbrial polypeptide constructs stabilized by donor strand complementation and connected via specific polypeptide linkers, conjugated to Campylobacter jejuni capsule polysaccharide or Shigella LPS. The construct covers multipartite fusions of subunits from different fimbrial types and includes refinements such as sequence specifics, signal peptides, linker compositions, and N-terminal deletions for optimized immunogenic efficacy.
Stated Advantages
Induces an immune response against multiple bacterial species including ETEC, Campylobacter jejuni, and Shigella spp. in a single composition.
Avoids potential autoimmune responses such as Guillain-Barré Syndrome by using Campylobacter jejuni capsule polysaccharide instead of lipooligosaccharide antigens.
Stabilizes ETEC fimbrial adhesins through donor strand complementation, enhancing antigen stability and immune recognition.
Enables broad immunogenic coverage by combining multiple fimbrial subunits from different ETEC fimbrial types into multipartite fusion constructs.
Provides a protein carrier function to the ETEC recombinant polypeptides, enhancing immunogenicity of polysaccharide antigens like capsule polysaccharide or lipopolysaccharide.
Can be synthesized and conjugated using established methods and demonstrated immunogenicity in animal models.
Documented Applications
Immunization of mammals, including humans, to induce immunity against enterobacteria such as Campylobacter jejuni, Escherichia coli (ETEC), and Shigella spp.
Vaccines targeting bacterial diarrhea caused by ETEC, Campylobacter jejuni, and Shigella species in populations including travelers and young children in resource-limited settings.
Use of recombinant polypeptide constructs conjugated to bacterial polysaccharide antigens as vaccine components.
Combined vaccines against multiple enteric pathogens to simplify prevention and reduce morbidity from diarrheal diseases globally.
Non-human primate and murine models for testing immunogenicity and efficacy of the conjugate vaccines.
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