Gallid alphaherpesvirus 3 (MDV-2), a viral vector against different avian pathogens: a new vaccination strategy in the poultry industry
AssigneeFarmacolóGicos Veterinarios S.A.C.
InventorsKatherine Ivette Calderón Mayo, Aldo Stanlee Rojas Neyra, Manolo Clemente Fernández Díaz
Life of this patent
Estimating the term…
Abstract
The present invention refers in general to the field of veterinary medicine and especially to the development of vectored vaccines using the Gallid alphaherpesvirus 3 (GaHV-3) or Marek's disease serotype 2 (MDV-2) vector, which contains protective antigens against different avian pathogens such as Newcastle disease virus (NDV), avian infectious laryngotracheitis virus (ILTV), infectious bursal disease virus (IBDV) or Gumboro disease virus, avian influenza virus (AIV), infectious bronchitis virus (IBV).
Claims15
A recombinant Gallid alphaherpesvirus 3 (GaHV-3;
- MDV-2) virus that expresses a heterologous polynucleotide encoding and expressing a gene or antigen from an avian pathogen, inserted into the UL45/UL46 intergenic region and the US2 gene regions of Gallid alphaherpesvirus 3 (GaHV-3;
- MDV-2).
An immunogenic composition characterized in that it comprises the recombinant virus of Gallid alphaherpesvirus 3 (GaHV3;
- MDV-2), which expresses the F gene of NDV genotype XII identified with SEQ ID NO. 5 and pharmaceutically acceptable excipients.
An immunogenic composition characterized in that it comprises the recombinant Gallid alphaherpesvirus 3 virus (GaHV3;
- MDV-2), which expresses glycoproteins D and I of ILTV strain VFAR-043 identified with SEQ ID NO.22 and pharmaceutically acceptable excipients.
An immunogenic composition characterized in that it comprises the recombinant Gallid alphaherpesvirus 3 virus (GaHV3;
- MDV-2), which expresses the VP2 gene of IBDV strain Faragher 52/70, identified with SEQ ID NO.36 and pharmaceutically acceptable excipients.
A procedure for generating a recombinant virus of Gallid alphaherpesvirus 3 (GaHV3;
- MDV-2), expressing antigenic genes against different avian diseases NDV, ILTV, IBDV, IBV, AIV, wherein the method comprises:
- a) Design and construction of gRNAs and donor plasmid;
- b) Generation of recombinants;
- c) Recovery of recombinant viruses;
- e) Selection and characterization of recombinant viruses containing the GFP cassette+the cassette of interest (protective antigen), by conventional PCR amplifying the complete inserted cassette and the joining zones between the genome of the recombinant virus obtained and the genome of the modified virus itself,
- f) Removal of the expression cassette of the fluorescent reporter gene “GFP” by the Cre-Lox system and selection of recombinant clones using conventional PCR;
- g) Detection of expression of proteins of interest in cells infected with recombinant viruses by indirect immunofluorescence (IIF);
- h) Determination of gene expression by Western blot (WB);
- i) Evaluation of genetic stability of the cassette inserted into the virus genome by conventional PCR and/or Western blot (WB);
- and
- j) Measure in vitro growth properties.
Dependencies are inferred from the claim text.