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Escape mutants of newcastle disease virus as marker vaccines

A Newcastle disease virus, anti-Newcastle disease virus technology, applied in the direction of virus antigen components, antiviral immunoglobulin, inactivation/attenuation, etc., can solve the problem of indistinguishable antibodies

Inactive Publication Date: 2006-03-08
ZOETIS W LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, antibodies elicited by current vaccinations against NDV immunogens are often indistinguishable from those found after infection with highly pathogenic and infectious NDV

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0044] Embodiment 1-experimental scheme

[0045] The following Phase 4 protocol briefly illustrates a useful route for developing marker vaccines against Newcastle Disease Virus.

[0046] Phase 1

[0047] It was determined whether natural infection with PMV1 (paramyxovirus serotype 1, identified as NDV) induces antibodies to the antigenic sites recognized by the mAb 54 antibody. Further analysis was performed to generate escape mutants lacking the antigenic site recognized by mAb 54 on the fusion protein.

[0048] section 2

[0049] Escape mutants lacking the antigenic site recognized by mAb 54 on the fusion protein were genetically identified.

[0050] Phase 3

[0051] The stability test of the escape mutant strain was carried out to check the status of the wild type or virulence back mutation.

[0052] Phase 4

[0053] Development and validation of assays to distinguish between anti-wild-type antibodies and anti-vaccine strain antibodies.

[0054...

Embodiment 2

[0078] The method of embodiment 2-selection escape mutant strain

[0079] The method described by Russell, P.H., J. Gen. Virol. (1984) 65, 795-798 may be used. These procedures are outlined below.

[0080] • A batch of monoclonal antibody 617 / 54 (mAb54) was prepared using 'Technomouse'.

[0081] ·The NDW strain was cultured with 9-day-old poultry embryos.

[0082] • Determination of the neutralization index of mAb54. Virus infectivity titers were determined by micro-neutralization assay with different dilutions of heat-inactivated mAb. Equal amounts (50 μl) of virus and mAb were reacted at 37°C for 2 hours, and then adsorbed into the microwells of VERO cells. Perform IIP test and count the number of infected cells. Expect virus infectivity to be reduced by 10 5 above.

[0083] ·Immune breeding of viral plaques. Equal amounts of virus and heat-inactivated mAb were incubated at 37°C for 2 hours. This mixture was diluted 1 / 10, 1 / 100 and 1 / 1000 with PBS / medium. E...

Embodiment 3

[0087] Efficacy studies of NDW escape mutants and 'POULVAC' vaccine

[0088] method

[0089] • Two vaccinated groups (one mutant and 'Poulvac' vaccine), each consisting of 25 day old chickens.

[0090] • Ten unvaccinated chicks served as the control group.

[0091] • The virus removed from the plaques was passaged six times with avian embryos for all inoculations of the mutant strains.

[0092] ·Each vaccinated chick is scheduled to receive 10 6..5 EID 50 .

[0093] Intramuscular injection of NDV (Herts / 33) with strong pathogenicity to the birds for attacking the virus, the dosage of each bird is 0.5ml10 5.0 ELD 50 .

[0094] Initially, the titer of the virus was determined by titrating the mutant strain. As shown in Table 1. The mutant strain p.13 was diluted to 10 6.5 EID 50 / 0.5ml and the total amount of 12.5ml is extensively sprayed to 25 chicks, and each is isolated and fed. The titer of each vial of 'Poulvac' vaccine is 10 10.02 EID 50 (provided by FortDo...

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Abstract

A vaccine against Newcastle Disease contains one or more mutant immunogens of the NDW strain. The mutant immunogen lacks the antigenic binding site on the F glycoprotein which is recognized by the monoclonal antibody mAb 54. Reagent kits and assay methods help to distinguish vaccinated members of a poultry flock from those that may have been infected with wild-type Newcastle Disease virus.

Description

field of invention [0001] The present invention relates to a novel anti-Newcastle disease vaccine and to a method of protecting poultry from Newcastle disease. In more detail, the present invention relates to a safe and effective marker vaccine against Newcastle disease, as well as kits and detection methods for testing poultry, so that poultry farmers can distinguish between vaccinated poultry and those that may have been infected in poultry flocks Poultry birds infected with wild-type Newcastle disease virus were differentiated. The invention also relates to novel mutant immunogens for use in vaccines against Newcastle disease. Background of the invention [0002] Newcastle disease (ND) is often a severe and fatal disease of poultry and can cause significant economic losses. The disease is caused by Newcastle disease virus (NDV), which belongs to the genus Paramyxovirus of the family Paramyxovirdae. Among avian paramyxoviruses, nine different serotypes have been identif...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/17C12N7/08C12N7/02C07K16/10
Inventor H·J·吉利斯I·H·布朗D·J·亚历山大M·S·科林斯
Owner ZOETIS W LLC
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