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Infectious bovine viral diarrhea virus

a bovine viral and virus technology, applied in the field of animal health, can solve the problems of the inability to fully absorb the virus, so as to reduce the symptoms of bvdv

Inactive Publication Date: 2007-01-18
ELBERS KNUT +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

"The invention is about a DNA molecule that contains a nucleotide sequence complementary to a BVDV RNA that causes infectious BVDV particles. When this DNA molecule is introduced into susceptible host cells, it can produce BVDV particles that are lethal to calves. The invention also includes an infectious BVDV clone generated by passaging a wild-type BVDV strain in cell culture. The DNA molecule can be used to attenuate BVDV strains by introducing mutations and collecting viral particles produced by these cells. The technical effect of the invention is the creation of a tool for research and development of a live vaccine for BVDV."

Problems solved by technology

Killed vaccines (inactivated whole virus) or subunit vaccines (conventionally purified or heterologously expressed purified viral proteins) are most often inferior to live vaccines in their efficacy to produce a full protective immune response even in the presence of adjuvants.
Live BVDV vaccines, although attenuated, are most often associated with safety problems.
Therefore, they cannot be applied to breeding herds that contain pregnant cows.
Furthermore, revertants of attenuated live BVDV pose a serious threat to cattle.
For conventionally derived attenuated viruses wherein the attenuation is achieved by conventional multiple passaging, the molecular origin as well as the genetic stability of the attenuation remains unknown and reversion to the virulent wild-type is unpredictable.

Method used

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  • Infectious bovine viral diarrhea virus
  • Infectious bovine viral diarrhea virus
  • Infectious bovine viral diarrhea virus

Examples

Experimental program
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Effect test

example 1

[0150] Materials & Methods

[0151] Cells and viruses. MDBK cells were obtained from the American Type Culture Collection (Rockville, Md.). Cells were grown in Dulbecco's modified Eagle's medium supplemented with 10% fetal calf serum (FCS; tested for the absence of pestivirus and antibodies against pestiviruses) and nonessential amino acids. Bovine viral diarrhea strain New York '93 (field isolate VLS#399) was kindly provided by E. J. Dubovi (New York State College of Veterinary Medicine, Cornell University, Ithaca). The virus underwent one animal passage and was designated “New York '93 / C” thereafter.

[0152] Infection of cells, immunofluorescence assay and virus peroxidase assay. Since pestiviruses are highly associated with their host cells, lysates of infected cells were used for reinfection of culture cells. Lysates were prepared by freezing and thawing cells 3 to 5 days after infection and were stored at −70° C. Unless indicated otherwise in the text, a multiplicity of infection ...

example 2

[0199] Experimental Design

[0200] Twelve pregnant heifers were selected from a BVDV negative herd. The following group of 5 / 7 heifers were included in the trial:

No.InoculationVirusGroup 1:5One i.n. administration,XIKE-A3 ml in each nostrilGroup 2:5One i.n. administration,NY-933 ml in each nostril

[0201] Heifers were moved to the experimental facilities 8 days before inoculations. Pregnancy status was confirmed after transport into the experimental facility. Heifers were between days 60 and 90 of gestation on the day of inoculation. Inoculation took place for all Is animals at one point of time with 2.5×104 TCID50 / ml of the respective virus applied in 6 ml tissue culture supernatant.

[0202] Heifers were monitored for the presence of clinical signs of BVDV infection including abortions during the observation period. The experiment was terminated 9 weeks after infection. Non-aborted cows were slaughtered, the uterus examined and collected. Foetal organ samples were collected during ro...

example 3

[0205] This study aimed to assess the efficacy of BVDV isolates against foetal infection. Efficacy of the NY93 infectious copy derivative BVDV recombinant (type II) with a deletion of the RNase function in the E(RNS) protein XIKE-B (H349Δ) is investigated to prevent fetal infection after an heterologous type I challenge.

[0206] Between day 60 and 90 is the most sensitive period for fetal exposure to BVDV. Therefore in this heifers derived from BVDV-free farm (and confirmed seronegative for BVDV) have been immunized by a single exposure with XIKE B (i.m.). Thereafter heifers were inseminated and between day 60-90 animals, when animals are supposed to be highly sensitive to BVDV fetal infection, a challenge infection with a wild type field virus was performed. The intranasal route for challenge was chosen as this mimics the normal route of infection in the field best.

[0207] Experimental Design:

[0208] Heifers were selected from a BVDV negative herd. The heifers were tested serologica...

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Abstract

The invention belongs to the field of animal health and in particular Bovine Viral Diarrhea Virus (BVDV). The invention provides infectious BVDV clones and methods to produce said BVDV clones. The invention further relates to methods of attenuating said clones, attenuated BVDV clones and vaccines comprising said attenuated clones.

Description

RELATED APPLICATION [0001] This application is a continuation of U.S. application Ser. No. 10 / 236,542, filed Sep. 6, 2002, which claims the benefit of priority to U.S. Provisional Application Ser. No. 60 / 322,974, filed Sep. 18, 2001, which are herein incorporated by reference.BACKGROUND OF THE INVENTION [0002] The invention belongs to the field of animal health and in particular Bovine Viral Diarrhea Virus (BVDV). The invention provides infectious BVDV clones and methods to produce said BVDV clones. The invention further relates to methods of attenuating said clones, attenuated BVDV clones and vaccines comprising said attenuated clones. [0003] Bovine Viral Diarrhea Virus (BVDV) is the causative agent of BVD and mucosal disease in cattle (Baker, J. C., 1987, J. Am. Vet. Med. Assoc. 190:1449-1458; Moennig, V. and Plagemann, J., 1992; Adv. Virus Res. 41:53-91; Thiel, H. J. et al., 1996, Fields Virology 1059-1073). Fetal infection during pregnancy can result in the resorption of the fet...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12Q1/70C12Q1/68C12N7/00C12N7/04
CPCA61K2039/5254A61K2039/53C12N2770/24362C12N2770/24361C12N7/00A61P37/04
Inventor ELBERS, KNUTMEYER, CHRISTIANEVON FREYBURG, MARTINAMEYERS, GREGOR
Owner ELBERS KNUT
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