Recombinant Non-Pathogenic Marek's Disease Virus Constructs Encoding Infectious Laryngotracheitis Virus and Infectious Bursal Disease Virus Antigens
AssigneeIntervet Inc.
InventorsStephanie Cook, Mohamad Morsey, Ian Tarpey, Iwan Verstegen, Paulus Jacobus Antonius Sondermeijer, Paul Vermeij
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Abstract
The present invention discloses novel recombinant multivalent non-pathogenic Marek's Disease virus constructs that encode and express both Infectious Laryngotracheitis Virus protein antigens and an Infectious Bursal Disease virus protein antigen, and methods of their use in poultry vaccines.
Claims13
-17. (canceled)
A method for the preparation of a vaccine, the method comprising:
- a. infecting a culture of chicken embryo fibroblast cells (CEFs) with a recombinant herpesvirus of turkeys (rHVT), wherein the rHVT comprises a first heterologous nucleic acid located in a first nonessential site in the rHVT genome and a second heterologous nucleic acid located in a second nonessential site in the rHVT genome;
- wherein the first heterologous nucleic acid comprises both a nucleotide sequence that encodes an Infectious Laryngotracheitis Virus glycoprotein D (ILTV gD) and a nucleotide sequence that encodes an Infectious Laryngotracheitis Virus glycoprotein I (ILTV gI);
- wherein the second heterologous nucleic acid comprises a nucleotide sequence that encodes an Infectious Bursal Disease Virus viral protein 2 (IBDV VP2);
- wherein the first nonessential site and the second nonessential site are either the same or different;
- and wherein when the first nonessential site and the second nonessential site are different, one is the US2 site and the other is the UL54.5 site;
- b. harvesting the infected CEFs;
- c. optionally, obtaining the rHVT from the CEFs;
- and
- d. admixing the harvested infected CEFs from step b., or the obtained rHVT from step c., with a pharmaceutically acceptable carrier.
A method for the preparation of a vaccine, the method comprising:
- infecting a culture of CEFs with a rHVT that comprises a recombinant nucleic acid comprising in 5′ to 3′ direction in the following order:
- (i) a murine cytomegalovirus immediate early 1 (mCMV-IE1) promoter;
- (ii) a coding sequence for Infectious Bursal Disease Virus viral protein 2 (IBDV VP2);
- (iii) a transcription terminator sequence;
- (iv) an Infectious Laryngotracheitis Virus glycoprotein D (ILTV gD) promoter;
- (v) a coding sequence for the ILTV gD protein;
- (vi) an Infectious Laryngotracheitis Virus glycoprotein I (ILTV gI) promoter;
- and
- (vii) a coding sequence for the ILTV gI protein.
A chicken embryo fibroblast (CEF) cell, the CEF comprising a recombinant herpesvirus of turkeys (rHVT) comprising a nucleotide sequence that encodes an Infectious Laryngotracheitis Virus glycoprotein D (ILTV gD), a nucleotide sequence that encodes an Infectious Laryngotracheitis Virus glycoprotein I (ILTV gI), and a nucleotide sequence that encodes an Infectious Bursal Disease Virus viral protein 2 (IBDV VP2);
- wherein the nucleic acid sequence is located in a nonessential site in the rHVT genome;
- and wherein the nonessential insertion site is selected from the group consisting of the US2 site and the UL54.5 site.
Dependencies are inferred from the claim text.