Recombinant Marek's disease virus (MDV) and vaccine

Drug – bio-affecting and body treating compositions – Antigen – epitope – or other immunospecific immunoeffector – Recombinant virus encoding one or more heterologous proteins...

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4242291, 4242141, 424816, 424 932, 435 693, 4353201, 536 2372, 536 241, A61K 3912, A61K 39245

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060132612

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BRIEF SUMMARY
TECHNICAL FIELD OF THE INVENTION

The present invention relates to a novel recombinant virus which can express an exogenous gene in an animal cell or in an animal body and can persistently infect even in the presence of a maternal antibody to continue antigen stimulation for a long period of time while escaping from challenge of the host immune system, thereby providing a vaccine which is effective even for conventional animals having a maternal antibody. The present invention also relates to a polyvalent live vaccine for animals using said recombinant virus. The present invention further relates to a recombinant virus vector which can be used as a drug delivery system (DDS) for production of physiologically active substances such as hormones or cytokines within the living body.


BACKGROUND OF THE INVENTION

Virus vector has widely been studied for efficiency of vaccination and for use as a gene introduction system into the living body. Particularly in the poultry industry, a study for application of poxviruses, especially fowlpox virus, has made a rapid progress. Under the circumstances, in order to develop a more efficient vector, the present inventors have investigated to make a vector from Marek's disease virus type 1 (hereinafter also referred to as "MDV1"), a kind of avian herpes viruses, and have reported many results thereof. For example, the present inventors have investigated more than 20 sites on the MDV1 genome, and as a result, have identified US10 gene as an insertion site for an exogenous gene which can stably retain an exogenous gene but does not impair the vaccine effects against Marek's disease [Japanese patent application No. 4-205933 (Japanese patent publication No. 6-22757); 4th International Symposium on Marek's Disease (1992), Amsterdam; Vaccine 1994 Vol. 12, 953-957]. A recombinant virus which incorporates F protein gene of Newcastle disease virus into the US10 gene exhibited sufficient effects as a vaccine in SPF chickens, said effects being persistent over at least 24 weeks after inoculation (4th International Symposium on Marek's Disease (1992), Amsterdam). This recombinant virus has been proved to show protective effects against both Marek's disease and Newcastle disease even in chickens having maternal antibodies but the protective effects were somewhat lowered than in SPF chickens. That is, this recombinant virus showed efficiently 100% protective effects against Newcastle disease in SPF chickens whereas it showed somewhat lowered effects, i.e. 70 to 90% level, in field chickens having maternal antibodies (Current Developments in the Molecular Biology of Marek's Disease Virus Workshop, 1995, Florida). Such a decrease in the effects was supposedly due to suppression of in vivo growth of the recombinant virus by maternal antibodies against Newcastle disease and Marek's disease when field chickens are immunized with the recombinant virus.


DISCLOSURE OF THE INVENTION

As mentioned above, when a live vaccine comprising a recombinant virus is inoculated into an animal having maternal antibodies, a problem arises that said virus is eliminated or viral growth is inhibited by the host immune system such as maternal antibodies. Thus, an improved recombinant virus is desired which retains the properties of a live vaccine and can continuously provide antigen stimulation for a long period of time while escaping from the challenge of the immune system.
In order to solve the above-mentioned problem, the present inventors have noted promoters derived from MDV1 and have cloned several putative promoter genes from Marek's disease virus (hereinafter also referred to as "MDV") genome. Using these promoter genes, the present inventors have conducted an experiment for expression of an exogenous gene in a recombinant virus comprising said genes. As a result, it was found, that among several promoter genes which the present inventors have cloned, a promoter from glycoprotein B (hereinafter referred to as "gB") gene, a gene homologous to herpes simplex virus gB gene, exhibited particularly excellen

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