Recombinant respiratory syncytial virus G protein fragments
Inventors
Khurana, Surender • Golding, Hana
Assignees
US Department of Health and Human Services
Publication Number
US-10377798-B2
Publication Date
2019-08-13
Expiration Date
2034-12-02
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Abstract
Compositions and methods useful for producing an immune response in a subject specific for the RSV G protein are described herein. The new methods and compositions described herein are made possible by the development of a new recombinant RSV G protein fragment, which has been engineered for in vitro production and is antigenically similar to the native RSV G protein. The recombinant RSV G protein fragment is capable of inducing the production of RSV G-specific antibodies when injected into a subject. These antibodies can recognize both RSV A and RSV B strains and inhibit infection of both viruses. Accordingly, the compositions and methods described herein may be useful in protecting subjects from RSV infection via immunization, raising antibodies specific for RSV, which can in turn be used to treat RSV infection.
Core Innovation
Human respiratory syncytial virus (RSV) is a major global health problem, infecting nearly everyone early in life and causing significant mortality and morbidity. Despite the public health need, no licensed vaccine or effective antiviral therapy exists for RSV. Existing vaccine approaches, such as inactivated vaccines, pose risks like enhanced disease upon natural infection, and live-attenuated vaccines are difficult to develop due to issues like moderate viral growth and attenuation reducing immunogenicity. Traditional vaccine development efforts have focused mainly on the RSV fusion (F) protein, while the attachment (G) protein was considered less attractive since most neutralizing antibodies target the F protein.
This invention addresses the need for an effective RSV vaccine by providing compositions and methods useful for producing an immune response specific for the RSV G protein. A new recombinant RSV G protein fragment has been engineered for in vitro production, which is antigenically similar to the native RSV G protein and capable of inducing production of RSV G-specific antibodies upon administration to a subject. These antibodies recognize both RSV A and B strains and inhibit infection of both viruses. This approach proposes that immunization with the recombinant RSV G protein fragment or administration of RSV G-specific antibodies could protect subjects from RSV infection or treat RSV infection.
The invention includes recombinant polypeptide fragments derived from the RSV G protein, particularly those that include only the ectodomain or subsections thereof, excluding portions of the transmembrane domain. For example, a polypeptide containing amino acids 67-298 of an RSV G protein is provided. These polypeptides can form oligomeric compounds with at least two copies of the polypeptide. The recombinant polypeptides can include additional sequences, such as epitope tags or cleavage sites, to facilitate tracking, purification, or enhance immunogenicity. Corresponding polynucleotides encoding these polypeptides, optimized for expression in various hosts including E. coli and mammalian cells, are also described. Methods for producing the recombinant RSV G polypeptides, as well as their use in generating an immune response in subjects against RSV G protein, are part of the innovation.
Claims Coverage
The patent contains multiple inventive features focusing on a soluble unglycosylated oligomeric RSV G ectodomain, its production, administration, and pharmaceutical compositions. Key inventive features pertain to the composition of the RSV G ectodomain, methods to generate immune response using it, and methods of producing the recombinant RSV G ectodomain in bacterial systems.
soluble unglycosylated oligomeric RSV G ectodomain
A composition comprising a soluble, unglycosylated, oligomeric form of the RSV G ectodomain, including specific amino acid sequences such as SEQ ID NO: 2. This forms the core antigenic component used in the invention.
method of generating an immune response to RSV G
Administering an amount of the soluble unglycosylated oligomeric RSV G ectodomain sufficient to elicit an immune response in a subject, with possible dosing ranges from 0.1 μg/kg to 100 μg/kg, optionally in combination with an adjuvant to enhance immunogenicity.
pharmaceutical composition comprising soluble unglycosylated oligomeric RSV G ectodomain
Pharmaceutical compositions containing the soluble unglycosylated oligomeric RSV G ectodomain, which can form homo-oligomers comprising at least two or four copies of the ectodomain, optionally including an adjuvant.
method of producing soluble unglycosylated oligomeric RSV G ectodomain in bacteria
A method involving expressing a nucleic acid encoding RSV G ectodomain in bacterial cells, lysing the cells to purify inclusion bodies, denaturing the protein, refolding it under redox conditions to obtain a soluble, unglycosylated, oligomeric form of RSV G ectodomain, and purifying the protein, where the nucleic acid may encode the amino acid sequence of SEQ ID NO: 2 and the nucleic acid sequence of SEQ ID NO: 1.
The claims cover the composition of a novel soluble unglycosylated oligomeric RSV G ectodomain, methods to use it for immunization, production techniques in bacterial systems, and associated pharmaceutical formulations. These features provide a comprehensive approach to creating effective RSV G protein based vaccines and therapeutics.
Stated Advantages
The recombinant RSV G protein fragment induces antibodies that recognize both RSV A and B strains and inhibit infection of both viruses.
The fragments can be produced efficiently in vitro, including in bacterial systems, facilitating scalable vaccine production.
The recombinant protein forms oligomers resembling native viral proteins, enabling generation of relevant conformational epitopes for neutralizing RSV.
Vaccination with the recombinant RSV G polypeptides provides protection against RSV infection and reduces viral pathology, including lung damage and cytokine induction.
Documented Applications
Immunization of subjects against RSV infection by administering the recombinant RSV G polypeptides or polynucleotides encoding them to elicit an immune response.
Raising RSV G protein-specific antibodies in subjects for use in treatment of RSV infection.
Production of recombinant RSV G protein ectodomains using bacterial expression systems followed by refolding under redox conditions for vaccine production.
Use of the recombinant RSV G ectodomain in pharmaceutical compositions for vaccination or therapeutic purposes.
Protection of mice and rabbits from RSV A and B strain challenge following vaccination with the recombinant RSV G protein, demonstrating cross-strain protective immunity.
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