Pseudorabies vaccines and DNA vectors for recombination with pox viruses

Claim 2

A recombinant pox virus capable of expressing in a host one or more immunogenic proteins of pseudorabies virus.

Claim 3

A recombinant pox virus of claim 1, which is of the species vaccinia.

Claim 4

A recombinant pox virus of claim 1, wherein the immunogenic protein is an envelope protein of pseudorabies virus.

Claim 5

A recombinant pox virus of claim 3, wherein the envelope glycoprotein of pseudorabies virus is the gp50 glycoprotein, the gII glycoprotein, the gIII glycoprotein or immunogenic subunits or fragments thereof.

Claim 6

A recombinant vaccinia virus containing, in a region of the viral genome nonessential for replication of the virus, one or more genes encoding the protein components of envelope glycoproteins of pseudorabies virus, the gene or genes being under control of individual vaccinia promoters.

Claim 7

A recombinant vaccinia virus containing, in a region of the viral genome nonessential for replication of the virus, one or more genes encoding the protein components of envelope glycoproteins of pseudorabies virus, the gene or genes being under control of individual vaccinia promoters.

Claim 8

A recombinant vaccinia virus of Claim 5, wherein the envelope glycoprotein is the gp50, gII, or gIII glycoprotein or immunogenic subunits or fragments thereof.

Claim 9

A vector for insertion of foreign DNA and for subsequent in vivo recombination with pox virus to produce a recombinant pox virus capable of expressing the foreign DNA in a host, com­prising; a. a pox virus promoter linked to; b. A DNA sequence containing a multiple cloning site for insertion of antigen encoding DNA, the cloning site being located such that the inserted DNA will be under control of the promoter; and; c. DNA sequences of the pox virus flanking the construct of elements a and b, the DNA sequences being substantially homologous to a region of the pox viral genome which is nonessential for replication of the pox virus; d. a replicon for replication in a prokaryo­tic host; and e. a structural gene encoding a marker or indicator for selection of prokaryotic hosts transformed with the vector, the gene being under control of a prokaryotic promoter.

Claim 10

A vector for insertion of foreign DNA and for subsequent in vivo recombination with pox virus to produce a recombinant pox virus capable of expressing the foreign DNA in a host, com­prising; a. a pox virus promoter linked to; b. A DNA sequence containing a multiple cloning site for insertion of antigen encoding DNA, the cloning site being located such that the inserted DNA will be under control of the promoter; and; c. DNA sequences of the pox virus flanking the construct of elements a and b, the DNA sequences being substantially homologous to a region of the pox viral genome which is nonessential for replication of the pox virus; d. a replicon for replication in a prokaryo­tic host; and e. a structural gene encoding a marker or indicator for selection of prokaryotic hosts transformed with the vector, the gene being under control of a prokaryotic promoter.

Claim 11

A vector of Claim 7, wherein the foreign DNA encodes a bacterial, mycobacterial or viral antigen.

Claim 12

A vector of Claim 7, for recombination with vaccinia virus, wherein the pox virus promoter is the 11K promoter, the 11KΔ5 promoter, the 7.5K promoter or the 30K promoter of vaccinia virus and where the flanking DNA sequences are sequences homologous to the thymidine kinase gene of vaccinia virus.

Claim 13

A vector of Claim 7, further comprising: f. a pox virus promoter linked to a struc­tural gene which encodes a marker or indicator for selection of recombinant pox virus, the promoter and structural gene being located between the flanking DNA sequences.

Claim 14

A vector of Claim 10, for recombination with vaccinia virus, wherein the pox virus promoter is the vaccinia BamF promoter and the struc­tural gene encoding a marker or indicator is the E. coli lacZ gene encoding B-galactosidase.

Claim 15

A vector for insertion of foreign DNA and for subsequent in vivo recombination with vaccinia virus to produce a recombinant vaccinia virus capable of expressing the foreign DNA in a host, comprising: a. a vaccinia promoter linked to; b. a DNA sequence containing a multiple cloning site for insertion of the antigen encoding DNA, the cloning site being positioned such that the inserted DNA will be under control of the vaccinia promoter; c. a vaccinia promoter linked to a structural gene encoding a marker or indicator for selection of recombinant vaccinia virus; d. DNA sequences of vaccinia virus flanking the construct of elements a-c, the DNA sequences being substantially homologous to a region of the vaccinia viral genome which is nonessential for replication of the virus; e. a replicon for replication in a pro­karyotic host cell; and f. a structural gene encoding a marker or indicator for selection of prokaryotic hosts transformed with the vector, the gene being under control of a prokaryotic promoter.

Claim 16

A vector for insertion of foreign DNA and for subsequent in vivo recombination with vaccinia virus to produce a recombinant vaccinia virus capable of expressing the foreign DNA in a host, comprising: a. a vaccinia promoter linked to; b. a DNA sequence containing a multiple cloning site for insertion of the antigen encoding DNA, the cloning site being positioned such that the inserted DNA will be under control of the vaccinia promoter; c. a vaccinia promoter linked to a structural gene encoding a marker or indicator for selection of recombinant vaccinia virus; d. DNA sequences of vaccinia virus flanking the construct of elements a-c, the DNA sequences being substantially homologous to a region of the vaccinia viral genome which is nonessential for replication of the virus; e. a replicon for replication in a pro­karyotic host cell; and f. a structural gene encoding a marker or indicator for selection of prokaryotic hosts transformed with the vector, the gene being under control of a prokaryotic promoter.

Claim 17

A vector of Claim 12, wherein the foreign DNA is DNA which encodes a bacterial, mycobacterial or viral antigen.

Claim 18

A vector of Claim 14, wherein: (i) the vaccinia promoter linked to a DNA sequence containing a multiple cloning site is the 7.5K, 11K, 11KΔ5 or 30K promoter of vac­cinia virus; (ii) the vaccinia promoter controlling the structural gene encoding a marker or indicator for selection of recombinant vaccinia virus is the vaccinia BamF promoter; (iii) the structural gene encoding a marker or indicator for selection of recombinant vaccinia virus is the E. coli lacZ gene; and (iv) the flanking DNA sequences are substan­tially homologous to the gene encoding thymi­dine kinase of vaccinia virus.

Claim 19

Plasmid vectors for insertion of antigen encoding DNA and for subsequent in vivo recom­bination with vaccinia virus to produce a recombinant vaccinia virus capable of expres­sing the antigen in a host, the vectors having the characteristics of pAbT752 and pAbT4007.

Claim 20

Plasmid vectors for insertion of antigen encoding DNA and for subsequent in vivo recom­bination with vaccinia virus to produce a recombinant vaccinia virus capable of expres­sing the antigen in a host, the vectors having the characteristics of pAbT752 and pAbT4007.

Claim 21

A recombinant pox virus produced by: a. inserting DNA encoding an immunogenic protein into the cloning site of a vector of claim 7 or 10; b. allowing the resulting vector containing the protein-encoding DNA to recombine with pox virus in vivo to produce a recombinant pox virus; and c. isolating the recombinant pox virus.

Claim 22

A recombinant vaccinia virus produced by: a. inserting DNA encoding an immunogenic protein into the cloning site of a vector of claim 13 or 14; b. allowing the resulting vectors containing the protein-encoding DNA to recombine with vaccinia virus in vivo to produce a recombinant vaccinia virus; and c. isolating the recombinant vaccinia virus.

Claim 23

A recombinant vaccinia virus according to claim 16 or 17, wherein the DNA encodes a bacterial, mycobacterial or viral antigen.

Claim 24

A vector of claim 7 or 12, containing inserted DNA which encodes an immunogenic bacterial, mycobacterial or viral protein.

Claim 25

A vector of claim 19, wherein the protein is an envelope protein of pseudorabies virus for example the gp50, gII or gIII glycoprotein or an immunogenic subunit or fragment thereof.

Claim 26

Plasmid vectors for in vivo recombination with vaccinia virus to produce a recombinant vac­cinia virus which expresses an immunogenic envelope glycoprotein of pseudorabies virus in a host, the plasmid vectors having the charac­teristics of pAbT756, pAbT4018, pAbT501 or pAbT502.

Claim 27

Plasmid vectors for in vivo recombination with vaccinia virus to produce a recombinant vac­cinia virus which expresses an immunogenic envelope glycoprotein of pseudorabies virus in a host, the plasmid vectors having the charac­teristics of pAbT756, pAbT4018, pAbT501 or pAbT502.

Claim 28

A vector for insertion of two or more foreign DNA sequences and for subsequent in vivo recombination with pox virus to produce a recombinant pox virus capable of expressing the DNA sequence, comprising: a. two or more pox virus promoters, each promoter linked to; b. a DNA sequence containing a multiple cloning site for insertion of a protein-­encoding DNA sequence, the cloning site being located such that the inserted DNA sequence will be under control of the promoter; and; c. DNA sequences of the pox virus flanking the construct of elements a and b, the DNA sequences being substantially homologous to a region of the pox viral genome which is nonessential for replication of the pox virus; d. a replicon for replication in a prokaryo­tic host; and e. a structural gene encoding a marker or indicator for selection of prokaryotic hosts transformed with the vector, the gene being under control of a prokaryotic promoter.

Claim 29

A vector for insertion of two or more foreign DNA sequences and for subsequent in vivo recombination with pox virus to produce a recombinant pox virus capable of expressing the DNA sequence, comprising: a. two or more pox virus promoters, each promoter linked to; b. a DNA sequence containing a multiple cloning site for insertion of a protein-­encoding DNA sequence, the cloning site being located such that the inserted DNA sequence will be under control of the promoter; and; c. DNA sequences of the pox virus flanking the construct of elements a and b, the DNA sequences being substantially homologous to a region of the pox viral genome which is nonessential for replication of the pox virus; d. a replicon for replication in a prokaryo­tic host; and e. a structural gene encoding a marker or indicator for selection of prokaryotic hosts transformed with the vector, the gene being under control of a prokaryotic promoter.

Claim 30

A vector of claim 22, wherein each foreign DNA sequence encodes a bacterial, mycobacterial or viral antigen.

Claim 31

A vector of claim 22 or 23, for recombination with vaccinia virus, wherein the pox viral promoters are selected from the group consisting of the 11K promoter, the 7.5K promoter, and the 30K promoter of vaccinia virus and modified ver­sions thereof, and wherein the flanking se­quences are sequences homologous to the thy­midine kinase gene of vaccinia virus.

Claim 32

A vector of claim 22, 23 or 24, further comprising: f. a pox viral promoter linked to a struc­tural gene which encodes a marker or indicator for selection of recombinant pox virus, the promoter and structural gene being located between the flanking DNA sequences.

Claim 33

A vector of claim 25, for recombination with vaccinia virus, wherein the pox virus promoter is the vaccinia virus promoter and the struc­tural gene encoding a marker or indicator is the E. coli lacZ gene encoding B-galacto­sidase.

Claim 34

Plasmid vectors for insertion of two sequences of protein-encoding DNA and for subsequent in vivo recombination with vaccinia virus to produce a recombinant vaccinia virus capable of expressing the antigens, the vectors having the characteristics of pAbT4026 and pAbT4027.

Claim 35

Plasmid vectors for insertion of two sequences of protein-encoding DNA and for subsequent in vivo recombination with vaccinia virus to produce a recombinant vaccinia virus capable of expressing the antigens, the vectors having the characteristics of pAbT4026 and pAbT4027.

Claim 36

A vector of claim 22 or 26 containing inserted DNA sequences, each sequence encoding different proteins of pseudorabies virus, the proteins being selected from gp50, gII, gIII proteins and immunogenic subunits or fragments thereof.

Claim 37

Plasmid vectors for recombination with vaccinia virus to produce a recombinant vaccinia virus capable of expressing two different glycopro­teins of pseudorabies virus, the vectors having the characteristics of pAbT503 and pAbT506.

Claim 38

Plasmid vectors for recombination with vaccinia virus to produce a recombinant vaccinia virus capable of expressing two different glycopro­teins of pseudorabies virus, the vectors having the characteristics of pAbT503 and pAbT506.

Claim 39

A vaccine for vaccinating a host animal against pseudorabies virus, comprising a recombinant pox virus capable of expressing an immunogenic protein or proteins of pseudorabies virus, said pox virus e.g. being a vaccinia virus capable of expressing the gp50, gII or gIII proteins or pseudorabies virus, and/or said pox virus is e.g. vAbT20, vAbT67, vAbT69, vAbT78A, vAbT90, vAbT93, vAbT54R, or mixtures thereof.

Claim 40

A vaccine for vaccinating a host animal against pseudorabies virus, comprising a recombinant pox virus capable of expressing an immunogenic protein or proteins of pseudorabies virus, said pox virus e.g. being a vaccinia virus capable of expressing the gp50, gII or gIII proteins or pseudorabies virus, and/or said pox virus is e.g. vAbT20, vAbT67, vAbT69, vAbT78A, vAbT90, vAbT93, vAbT54R, or mixtures thereof.

Claim 43

A method of claim 31, for producing antigens against the gp50, gII or gIII glycoproteins of pseudorabies virus, wherein the cells are infected with a recombinant vaccinia virus capable of expressing the gp50, gII or gIII proteins pseudorabies virus or immunogenic fragments of subunits thereof.

Claim 48

Recombinant vaccinia viruses vAbT20, vAbT67, vAbT69, vAbT93, vAbT54R, vAbT78A, and vAbT90.

Claim 49

Recombinant vaccinia viruses vAbT20, vAbT67, vAbT69, vAbT93, vAbT54R, vAbT78A, and vAbT90.

Claim 50

Plasmids pAbT501, pAbT502, pAbT503, pAbT506, pAbT771 and pAbT4018.

Claim 51

Plasmids pAbT501, pAbT502, pAbT503, pAbT506, pAbT771 and pAbT4018.

Claim 52

Use of recombinant pox, e.g. vaccinia, viruses to combat diseases, e.g. Aujeszky's disease, caused by pseudorabies virus.

Claim 53

Use of recombinant pox, e.g. vaccinia, viruses to combat diseases, e.g. Aujeszky's disease, caused by pseudorabies virus.

Claim 54

Use of recombinant pox, e.g. vaccinia, viruses in the manufacture of vaccines against diseases, e.g. Aujeszky's disease, caused by pseudorabies virus.

Claim 55

Use of recombinant pox, e.g. vaccinia, viruses in the manufacture of vaccines against diseases, e.g. Aujeszky's disease, caused by pseudorabies virus.