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Recombinant Hvt Vectors Expressing Antigens Of Avian Pathogens And Uses Thereof

一种载体、禽类的技术，应用在病毒抗原成分、病毒/噬菌体、重组DNA技术等方向，能够解决未报导病毒载体等问题

Pending Publication Date: 2016-09-07
BOEHRINGER INGELHEIM ANIMAL HEALTH USA INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, no studies using SB-1 as a viral vector to express foreign protective genes have been reported

Method used

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  • Recombinant Hvt Vectors Expressing Antigens Of Avian Pathogens And Uses Thereof
  • Recombinant Hvt Vectors Expressing Antigens Of Avian Pathogens And Uses Thereof
  • Recombinant Hvt Vectors Expressing Antigens Of Avian Pathogens And Uses Thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Example 1 Construction of recombinant vHVT114 expressing NDV-F

[0074] Preparation of donor plasmid pHM103+Fopt

[0075] The intergenic I arm containing HVT FC126 was digested with NotI (see figure 2 ), the SV40 promoter and the plasmid pHM103 (Merial Limited) of SV40 poly A were dephosphorylated, and the 5.6 kb fragment was extracted by gel. A NotI-flanked 1.7 kb fragment of the chemically synthesized codon-optimized genotype VIId NDV-F gene (SEQ ID NO: 1, encoding SEQ ID NO: 2) was also subjected to NotI digestion followed by gel extraction of the 1.7 kb fragment. fragment. The 5.6 and 1.7 kb fragments were ligated to generate pHM103+Fopt ( image 3 ).

[0076] Production of Recombinant HVT Viral Vectors

[0077] In vitro recombination (IVR) was performed by co-electroporation of secondary chicken embryo fibroblasts (2° CEF cells) using pHM103+Fopt (as donor plasmid) and viral DNA isolated from HVT strain FC126. Use 1x10 in 300ul Opti-MEM 7 Co-electropora...

Embodiment 2

[0100] Example 2 Construction of recombinant vHVT110, vHVT111, vHVT112, vHVT113 and vHVT116 expressing NDV-F

[0101] Production and characterization of HVT recombinants vHVT110, vHVT111, vHVT112, vHVT113 and vHVT116 were performed essentially in the same manner as described in Example 1 for vHVT114. Table 4 shows the features surrounding the expression cassette that are unique to each construct, including the respective sequences.

[0102] Table 4

[0103] Characterization of the expression cassettes of individual HVT recombinants

[0104] name

parent virus

Promoter

F gene

Poly-A

loci

vHVT039

HVT

MDV

Wtnm-Texas

SV40

IG1

vHVT110

HVT

mCMVIE

Wt-VIId

SV40

IG1

vHVT111

HVT

SV40

Wt-VIId

SV40

IG1

vHVT112

HVT

MCMV-IE

Wt-YZCQ

SV40

IG1

vHVT113

HVT

MCMV-IE

Wt-Texas

SV40

IG1

vHVT114

HVT

SV40

Opt-VIId

SV4...

Embodiment 3

[0122] Example 3 Construction of vHVT306 (dual HVT vector expressing NDV-F and IBDV VP2)

[0123] A donor plasmid pHVT US2 SV-Fopt-synPA containing the SV40 promoter, synthetic NDV F codon-optimized VII gene, synthetic polyA tail and SORF3 and US2 arms flanked by HVT FC126 was constructed.

[0124] Production of recombinant virus

[0125] Co-electroporation of secondary CEF cells was performed using donor plasmid pHVT US2 SV-Fopt-synPA and viral DNA isolated from vHVT13 (HVT vector expressing IBDV VP2 gene, Merial Limited) following standard homologous recombination manipulations. Recombinants were produced essentially as described in Example 1 for vHVT114, plaque purified and recombinants identified by immunofluorescence.

[0126] After 5-plaque plaque purification, pure recombinant virus (vHVT306) was isolated, the purity of vHVT306 was tested, and the purity was confirmed by IFA and PCR.

[0127] PCR analysis

[0128] Viral DNA was extracted from vHVT306 main plate p...

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PUM

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Abstract

The present invention provides recombinant hvt vectors expressing antigens of avian pathogens and uses thereof, recombinant herpesvirus of turkeys (HVT) vectors that contain and express antigens of avian pathogens, compositions comprising the recombinant HVT vectors, polyvalent vaccines comprising the recombinant HVT vectors and one or more wild type viruses or recombinant vectors. The present invention further provides methods of vaccination against a variety of avian pathogens and method of producing the recombinant HVT vectors.

Description

[0001] This application is a divisional application of an invention patent application with the application number 201280066323.X, the application date is November 29, 2012, and the invention title is "Recombinant HVT vector expressing an antigen of an avian pathogen and its use". [0002] Cross Reference Related Applications [0003] This application claims priority to US Provisional Application 61 / 564,877, filed November 30, 2011 and US Provisional Application 61 / 694,957, filed August 30, 2012. technical field [0004] The present invention relates to recombinant viral vectors for insertion and expression of foreign genes for use as safe immune vehicles for protection against infection by a variety of pathogenic agents. It also relates to multivalent compositions or vaccines comprising one or more recombinant viral vectors for protection against infection by various pathogens. The present invention relates to methods of making and using recombinant viral vectors. Backgrou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/295A61K39/17A61K39/12A61P31/14C12N15/869
CPCA61K39/12C12N15/86A61K2039/70A61K2039/5256C12N2720/10034C12N2710/16043C12N2760/18134C12N2710/16034C12N2720/00034C12N2710/16041C12N2710/16071A61K2039/54A61K2039/552C12N2710/16343A61P31/12A61K39/15A61K39/17A61K39/255A61K39/295C07K14/03C07K14/125C07K14/14C12N15/63C12N15/869
Inventor M·巴布洛特T·梅巴特森J·普利特查德P·林茨
Owner BOEHRINGER INGELHEIM ANIMAL HEALTH USA INC
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