Vaccine for prevention of necrotic enteritis in poultry
Inventors
Roland, Kenneth • Diamos, Andrew • Mason, Hugh
Assignees
Arizona State University ASU • Arizona State University Downtown Phoenix campus
Publication Number
US-11857613-B2
Publication Date
2024-01-02
Expiration Date
2038-07-02
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Abstract
In certain embodiments, the present invention provides a poultry vaccine comprising an antigenic protein comprising a PlcC protein unit that is operably linked to a peptide linker that is operably linked to a NetB protein unit, where the vaccine is effective in stimulating a protective cellular and/or humoral immune response to C. perfringens. Methods are also provided for making the vaccine and for vaccinating poultry by administering such a vaccine.
Core Innovation
The invention provides a poultry vaccine comprising an antigenic protein that consists of a PlcC protein unit operably linked to a peptide linker, which is in turn operably linked to a NetB protein unit. This fusion protein is engineered to stimulate a protective cellular and/or humoral immune response to Clostridium perfringens in poultry. The antigen can be produced using recombinant expression in plants, where the fusion protein is either purified for use as an injectable vaccine or incorporated into feed for oral vaccination of poultry.
The problem addressed by this invention is the increasing incidence and economic impact of necrotic enteritis (NE) in poultry, primarily caused by C. perfringens. Previous reliance on Antimicrobial Growth Promoters (AGP) and coccidiostats has diminished due to public health concerns over antibiotic resistance, leading to a resurgence of NE. Existing vaccines have often provided incomplete protection, particularly as there was no commercial vaccine including a NetB immunogenic component prior to this invention.
This invention combines two of the most potent protective antigens against necrotic enteritis, namely the non-toxic C-terminal fragment of alpha-toxin (PlcC) and the NetB toxin, into a single fusion protein. The NetB portion can include one or more specific point mutations to reduce toxicity while preserving immunogenicity. The fusion protein is expressed strongly in plants, which lack cholesterol and are unaffected by the toxic effects of NetB, enabling high-yield production. The vaccine is formulated for protection through various administration routes, including injection, in-feed, or in ovo delivery to poultry.
Claims Coverage
The patent claims one main inventive feature in its independent claim, relating to a method for protecting an avian species from C. perfringens infection by administering a specific vaccine composition.
Vaccine with PlcC-linker-NetB fusion antigen for avian species
A method of protecting an avian species from C. perfringens infection comprising administering a vaccine that contains an antigenic protein. The antigenic protein consists of: - A PlcC protein unit operably linked at its C-terminus to the N-terminus of a peptide linker. - The peptide linker operably linked at its C-terminus to the N-terminus of a NetB protein unit. - The peptide linker has at least 95% sequence identity to SEQ ID NO: 4. - The claim is further supported by dependent claims that specify sequence identity for the PlcC and NetB units, allow particular substitutions within NetB to reduce toxicity, and describe the inclusion of additional peptide tags or signal sequences (e.g., 6His tag, plant signal peptide), as well as multiple possible administration routes (in-feed, injection, in ovo).
The inventive feature centers on a fusion vaccine antigen combining PlcC and NetB domains via a defined peptide linker, and methods of use for protecting avian species from C. perfringens infection, specifying key protein sequences and delivery methods.
Stated Advantages
Provides robust protective immunity against necrotic enteritis in poultry by combining both PlcC and NetB antigenic components into a single vaccine.
Enables vaccine production in plants, allowing either purified injectable preparations or cost-effective oral delivery through animal feed.
Avoids the need for antimicrobial growth promoters, supporting the goal of antimicrobial-free poultry farming.
Facilitates high expression and accumulation of the vaccine antigen in plants, unaffected by NetB toxicity.
Eliminates reliance on antibiotics for control of C. perfringens and associated diseases.
Documented Applications
Vaccination of poultry (including chicken and turkey) against necrotic enteritis and infections caused by C. perfringens, by administration of the described vaccine.
Administration via poultry feed for oral immunization.
Administration by injection for active immunization or maternal antibody transfer to offspring.
Administration in ovo to poultry embryos to provide early protection.
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