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BR-112017002577-B1 - Recombinant vaccine, mother seed of a recombinant avian adenovirus, and use of an avian adenovirus vector.

BR112017002577B1BR 112017002577 B1BR112017002577 B1BR 112017002577B1BR-112017002577-B1

Abstract

RECOMBINANT VACCINE, MOTHER SEED OF A RECOMBINANT AVIAN ADENOVIRUS AND USE OF AN AVIAN ADENOVIRUS VECTOR. A recombinant vaccine comprising an avian adenovirus vector, serotype 9 (FAdV-9), with at least one exogenous nucleotide sequence inserted encoding at least one antigen of a disease of interest and replacing the non-essential region of the adenovirus genome, and a pharmaceutically acceptable vehicle, adjuvant and/or excipient, wherein the at least one exogenous nucleotide sequence encoding at least one antigen of a disease of interest and replacing the non-essential region of the adenovirus genome is located between nucleotides 491 and 2782. The vector of this vaccine is stable for industrial-scale production. Similarly, even when this vaccine is administered in combination with a vaccine against Marek's disease, both vaccines produce an adequate immune response that is not affected by interference between them. Likewise, the efficacy of the recombinant vaccine is not affected by maternal antibodies and is able to induce an early and long-lasting protective response, even with a single application.

Inventors

  • Bernardo LOZANO-DUBERNARD
  • Ernesto Soto-Priante
  • David Sarfati-Mizrahi

Assignees

  • GRUPO INDUSTRIAL PECUARIO, S.A. DE C.V

Dates

Publication Date
20260310
Application Date
20150806
Priority Date
20140808

Claims (14)

  1. 1. Recombinant vaccine, characterized by comprising an avian adenovirus vector, serotype 9 (FAdV-9), having at least one inserted exogenous nucleotide sequence encoding at least one antigen of a disease of interest and replacing the non-essential region of the adenovirus genome, and a pharmaceutically acceptable vehicle, adjuvant and/or excipient, wherein the FAdV-9 vector is live, and comprises the SEQ ID No. 1 sequence, and the exogenous nucleotide sequence is located between nucleotides 491 and 2782.
  2. 2. Recombinant vaccine, according to claim 1, characterized in that the exogenous nucleotide sequence encodes at least one antigen of a disease of interest, selected from avian influenza, infectious laryngotracheitis of birds, Newcastle disease, bursa of Fabricius infection, infectious bronchitis, metapneumovirus disease (MPNV), Marek's disease, infectious anemia of birds or any other gene whose size allows insertion into the FAdV-9 vector.
  3. 3. Recombinant vaccine, according to claim 2, characterized in that the exogenous nucleotide sequence encodes an antigen from the group comprising hemagglutinin (HA) of avian influenza virus; glycoprotein B (gB) and glycoprotein D (gD) of avian infectious laryngotracheitis virus (ILT); HN and F proteins of Newcastle disease virus; VP2 protein of bursa of Fabricius infection virus; S1 and S2 proteins of infectious bronchitis virus; F protein of metapneumovirus (MPNV); and VP1, VP2 and VP3 proteins of avian infectious anemia.
  4. 4. Recombinant vaccine, according to claim 3, characterized in that the HA of the avian influenza virus is selected from at least one of the H1, H2, H3, H5, H6, H7 or H9 subtypes of said protein.
  5. 5. Recombinant vaccine, according to claim 1, characterized in that the exogenous nucleotide sequence comprises a gene encoding at least one antigen of an avian adenovirus other than FAdV-9 used as a viral vector.
  6. 6. Recombinant vaccine, according to claim 1, characterized in that the required virus concentration is at least 105.0% CCDI50% per dose.
  7. 7. Recombinant vaccine, according to claim 6, characterized in that the required virus concentration is at least 106.0% CCDI50% per dose.
  8. 8. Recombinant vaccine, according to claim 1, characterized in that, when the vaccine is live (active), the pharmaceutically acceptable vehicles are aqueous solutions, selected from the group consisting of aqueous solutions comprising trehalose, phosphate, glutamate, albumin (TPGA) stabilizer; aqueous solutions comprising peptone stabilizer; and aqueous solutions comprising skim milk.
  9. 9. Recombinant vaccine, according to claim 1, characterized in that the vaccine is prepared to be administered intramuscularly, intranasally, subcutaneously, by spray, nebulization, orally, or via drinking water, or in ovo.
  10. 10. Recombinant vaccine, according to claim 1, characterized in that it comprises a vaccine with whole Marek's disease virus.
  11. 11. Recombinant vaccine, according to claim 1, characterized in that it comprises an FAdV-9 vector having at least two inserted exogenous nucleotide sequences encoding at least two antigens of the same disease of interest or of different diseases of interest, and which replace the non-essential region of the adenovirus genome located between nucleotides 491 and 2782.
  12. 12. Recombinant vaccine, according to claim 1, characterized in that it comprises at least one second vaccine based on an FAdV-9 vector with at least one inserted exogenous nucleotide sequence encoding at least one antigen different from that of the first vaccine, either from the same disease of interest or from a different disease, and which replaces the non-essential region of the adenovirus genome located between nucleotides 491 and 2782.
  13. 13. Mother seed of a recombinant avian adenovirus, serotype 9 (FAdV-9), characterized by having an inserted exogenous nucleotide sequence that encodes an antigen of a disease of interest and that replaces the non-essential region of the adenovirus genome, located between nucleotides 491 and 2782, the sequence being obtained after 6 to 11 passages in cell culture.
  14. 14. Use of an avian adenovirus vector, serotype-9 (FAdV-9), characterized by having an inserted exogenous nucleotide sequence that encodes an antigen of a disease of interest and replaces the non-essential region of the adenovirus genome located between nucleotides 491 and 2782, to produce a recombinant vaccine.

Description

FIELD OF THE INVENTION [001] The present invention relates to vaccines based on viral vectors and, more particularly, relates to a vaccine based on a recombinant avian adenovirus vector, serotype-8. FUNDAMENTALS OF THE INVENTION [002] Adenoviruses are linear, double-stranded DNA viruses, 70-90 nm in diameter, non-enveloped, and with a capsid in the form of an icosahedron, which is formed by 240 hexons, 12 pentons, and fibers that extend from each vertex of the icosahedron. These hexons, pentons, and fibers represent the main antigens of the adenovirus, and those that determine its serotype. [003] The adenovirus genome is about 30-45 kb in size and has four early regions (E1, E2, E3 and E4) and five late regions (L1-L5). [004] Adenoviruses have been isolated from different species, with the two main genera being Aviadenovirus, isolated from birds, and Mastadenovirus, isolated from mammals. [005] Adenoviruses are considered good candidates for recombinant vectors for vaccine production because they are highly infectious and many of them are non-pathogenic. Furthermore, adenovirus vectors can effectively translate large genes and generate a prolonged immune response in animals. However, it is also known that the specific structure of adenoviruses requires the study of specific heterologous gene insertion sites for each species, and that it is practically impossible to generalize the biological behavior of the various known adenoviruses when they are being converted into a recombinant viral vector. [006] Until now, human, simian, avian, swine, and other adenoviruses have been used as vectors for potential use as recombinant vaccines. In particular, avian adenoviruses (FadV) are known to be potential candidates for the development of recombinant vaccines. However, as will be described below, the possibility of successfully using them on a commercial level, particularly in the veterinary industry, has generally been inhibited due to the inability to achieve their large-scale production with the necessary stability, since adenovirus production is carried out from cell lines whose yields are usually very poor, or nature prevents the achievement of viral vector stability in successive passages. Thus, it is easy to understand that, currently, there are no vaccines vectored in avian adenoviruses. [007] For example, International Publication No. WO94/24268 describes recombinant FAdV vectors with at least one heterologous nucleotide sequence inserted that are useful in generating an immune response in disease-susceptible birds. According to this document, the non-essential regions of the adenoviral genome that may be suitable for the heterologous gene to be replaced or inserted are those located at the right-hand end of the genome, preferably the region located between 0.0038 and 0.0039 in (97 and 99.9 m.u). The recombinant vaccine, according to this document, could apparently be used in combination with vaccines against other viruses, such as Marek's disease or Newcastle disease, although there is no evidence of this. Furthermore, this document does not illustrate the behavior of the adenovirus obtained after successive passages in cell lines for large-scale production. [008] Furthermore, US Patent No. 6,296,852 describes FAdV serotype-9 (FAdV-9) vectors, in which heterologous nucleotide sequences are inserted into non-essential regions of the viral genome. These regions may be non-coding regions located at the left and/or right end of the genome, preferably in the region located at the right end (3'), between 0.0023 and 0.0039 in (60 and 100 m.u). As for document WO94/24268, although this patent identifies another, broader region for the potential insertion of exogenous genes, this does not illustrate its behavior and, particularly, its integrity after successive passages in cell lines. [009] In another paper (Corredor, J. C. and Nagy, E., The non-essential region at the left end of the avian adenovirus 9 genome is suitable for foreign gene insertion/substitution, Virus Research. Vol. 149, 167-174; 2010). It is reported that the non-essential region at the 5' end of the FAdV-9 genome could also be a suitable site for insertion or substitution by exogenous genes to produce recombinant vectors. For example, one such vector was obtained by replacing the non-essential region located between nucleotides 491-2782 with the gene encoding an enhanced green fluorescent protein (EGFP). However, this paper, like the others, does not show whether the viral vectors as constructed would be stable or not when an exogenous gene encoding a disease antigen is inserted into them and, by replicating them in suitable cellular systems, a recombinant vaccine is obtained, since the paper only shows expression of the reporter gene and lacks in vivo assays; Furthermore, it doesn't mention the composition or the levels of protection against a disease of interest either. [0010] As can be seen, it is known that non-essential regions of the FAdV ge