Drug – bio-affecting and body treating compositions – Antigen – epitope – or other immunospecific immunoeffector – Recombinant virus encoding one or more heterologous proteins...
Reexamination Certificate
1997-09-04
2001-05-01
Salimi, Ali R. (Department: 1643)
Drug, bio-affecting and body treating compositions
Antigen, epitope, or other immunospecific immunoeffector
Recombinant virus encoding one or more heterologous proteins...
C424S229100, C424S813000, C424S205100, C435S320100, C435S235100, C435S069300, C435S005000, C536S023720
Reexamination Certificate
active
06224878
ABSTRACT:
FIELD OF THE INVENTION
The subject of the present invention is segments of the genome of the infectious bovine rhinotracheitis herpes-virus (IBR), contained in its short unique region, and encoding especially the g{dot over (I)} glycoproteins. The present invention relates to the DNA segments containing the gene encoding these antigenic glycoproteins and containing potential promoter sequences up to 400 nucleotides in 5′ of each gene, which segments are useful as possible source of BHV-1 promoters, for the expression of the genes for producing this and other glycoproteins, such as gE which in turn can be used to induce antibodies recognizing them, as potential sites for the insertion of foreign genes, and as potential sites for deletion or insertion in order to prevent the synthesis of these antigenic glycoproteins by the virus.
The subject of the invention is especially an insertion region in the genomic DNA of the bovine herpesvirus (BHV) comprising three open reading frames located in the short unique region (Us) of the genome corresponding to the nucleotide sequence SEQ ID No:1 of FIG.
2
.
Such an insertion region may comprise, for example the DNA sequence delimited by nucleotides 172 and 1311 of the sequence presented in FIG.
2
.
Another insertion region may comprise the DNA sequence delimited by nucleotides 1594 and 3318 of the sequence presented in FIG.
2
.
The subject of the invention is also plasmids comprising the insertion region according to the invention.
The plasmids according to the invention may also contain a foreign gene, isolated from an agent pathogenic for bovines, inserted in the said insertion region.
The said foreign gene is preferably chosen from a group essentially comprising the bovine coronavirus, the bovine rotavirus, the bovine leukosis virus, the foot-and-mouth virus, the mucosal disease virus, Salmonella,
E. coli
, where the said foreign gene is placed under the control of a promoter capable of directing its expression.
The subject of the present invention is particularly mutants of the IBR (BHV-1) virus containing mutations caused by total or partial deletion and/or by insertion in the said region and especially in the g{dot over (I)}-encoding gene, or the surrounding non-coding sequences, such that there is no longer any expression of the glycoprotein encoded by the gene which has been mutated or rendered inactive. Consequently, animals vaccinated with these mutants do not develop antibodies against the viral glycoprotein and can be serologically distinguished from animals infected by field BHV-1 strains and other vaccinal strains currently used.
The subject of the present invention is also the use of the mutant BHV-1 viruses described above as vectors for expression of foreign genes to prepare vaccines against bovine (ruminant) diseases. As above, animals vaccinated with these expression vectors do not develop antibodies against the viral glycoprotein and can be serologically distinguished from animals infected by field BHV-1 strains.
The recombinant BHV viruses may comprise a promoter for the insertion region, which promoter is functional in a host cell for the expression of the said foreign gene. The recombinant BHV viruses are designed so that at least one polypeptide encoded by the said foreign gene and at least one polypeptide encoded by the said BHV genomic DNA are expressed.
The subject of the present invention is furthermore the methods for producing the deleted mutants, the methods for producing the expression vectors and the methods for using the vaccines prepared from the deleted viruses or from the expression vectors expressing foreign genes.
The subject of the invention is also the vaccines comprising a recombinant BHV virus according to the invention. The vaccines may consist of the live virus, or of inactivated viral particles, or of subunits of the particles, with, preferably, a conventional adjuvant.
The subject of the present invention is also the expression of the BHV-1 g{dot over (I)} gene in various expression systems in order to use the expression products obtained as antigens. The present invention also relates to the preparation of serological reagents from the BHV-1 g{dot over (I)} antigens alone or with BHV-I gE antigens.
The subject of the present invention is furthermore the use of the BHV-1 g{dot over (I)} and optionally BHV-1 gE antigens to detect the presence of specific antibodies in the serum of bovines infected by BHV-1.
The present invention finally describes, for the first time, a 4190 bp DNA sequence contained in the Us region of the BHV-1 genome and encoding the genes homologous to HSV-1 g{dot over (I)}, HSV-1 gE and HSV-1 US9.
BACKGROUND OF THE INVENTION
Infectious bovine rhinotracheitis (IBR) is an acute and contagious infectious bovine disease, often aggravated by bacterial complications (Pastoret P. P., Ann. Med. Vet., 122, 371-391, 1978). This disease occurs, either in a respiratory form (IBR proper), which is predominant and which mainly affects young bovines, or in a genital form, infectious pustular vulvovaginitis (IPV) which has been historically important but which is currently more rare (Yates W. P., Can. J. Comp. Med., 46, 225-263, 1982). IBR is one of the principal causes of respiratory ailments in bovines and causes considerable economic losses both in the dairy industry and in meat production. Given the economic importance of this disease, sanitary barriers aimed at banning the import of infected animals have been introduced by a number of countries.
This disease is of viral nature and its etiological agent has been identified as type 1 bovine herpesvirus (BHV-1) (Madin S. H. et al., Science, 124, 721-722, 1956). This virus is classified in the family Herpesviridae and belongs more specifically to the subfamily Alphaherpesviridae of which the prototype is type 1 human herpes simplex virus (HSV-1) (Armstrong J. A. et al., Virology, 14, 276-285, 1961; Roizman B. et al., Arch. Virol., 123, 425-449, 1992). As most herpesviruses, the BHV-1 virus can be present in the latent state in its host and be reactivated under certain circumstances (stress, transport and the like). The control of this disease is based both on sanitary prophylaxis and on medical prophylaxis.
The clinical diagnosis is rendered difficult in the serious superinfected forms, which show, in this case, similarities with other bovine respiratory infections, and is not always easy to implement. Definite proof of an IBR disease can be provided only with laboratory tests (Gilbert Y. et al., Rev. Med. Vet., 3, 383-389, 1976; Fedida H. and Dannacher G., Bull. Lab. Vet., 5, 35-46, 1982). The isolation of the virus during cell culture from ecouvillonnages produced on the suspected animals remains the best means of diagnosing the disease. But this technique is slow and in particular cumbersome to implement. Consequently, simpler and more rapid techniques based on the detection of the BHV-1 antigens or, increasingly, on the detection of specific antibodies in the serum of the suspected animals, are now preferred in its place. Numerous tests using the ELISA technique are currently on the market (Cheng-Feng Z. and Forbes S. D., New Zealand, Vet. J., 36, 204-205, 1988). Seroneutralization remains, however, the official technique during European or international commercial transactions.
The control of IBR is based mainly on vaccination (Straub O. C. and Mawhinney I. C., Vet. Rec., 122, 407-411, 1988).
Three types of vaccines have been used to protect bovines against IBR (Kit M. et al., European Patent Application No. 0 316 658 A1, 1988).
The vaccines based on attenuated viruses obtained by a large number of successive passages of the pathogenic viruses on cells of bovine origin, by adaptation of these same viruses on cells of an origin other than bovine, or by selection of heat-stable spontaneous viral mutants. These vaccines are easy to produce, of relatively low cost and injectable.
They induce a rapid protection of long duration (Casselberry N. H., J.A.V.M.A., 152, 853-856, 1968). They have, nevertheless, a number of disadvantage
Audonnet Jean-Christophe Francis
Legastelois Isabelle Christine Marie-Andrée
Leung-Tack Patricia
Riviere Michel Emile Albert
Larson & Taylor
Merial
Salimi Ali R.
LandOfFree
Mutants and vaccines of the infectious bovine... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Mutants and vaccines of the infectious bovine..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Mutants and vaccines of the infectious bovine... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2522128