Drug – bio-affecting and body treating compositions – Antigen – epitope – or other immunospecific immunoeffector – Recombinant virus encoding one or more heterologous proteins...
Reexamination Certificate
1998-07-10
2001-09-25
Salimi, Ali R. (Department: 1648)
Drug, bio-affecting and body treating compositions
Antigen, epitope, or other immunospecific immunoeffector
Recombinant virus encoding one or more heterologous proteins...
C424S232100, C424S202100, C424S221100, C435S320100, C435S235100, C536S023720
Reexamination Certificate
active
06294176
ABSTRACT:
Within this application several publications are referenced by arabic numerals within parentheses. Full citations for these references may be found at the end of the specification immediately preceding the Sequence Listing. The disclosures of these publications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art as known to those skilled therein as of the date of the invention described and claimed herein.
BACKGROUND OF THE INVENTION
Raccoonpox virus (RPV) belongs to the orthopoxvirus family, like vaccinia virus, but has an advantage in that it is naturally attenuated and appears to be native to North America. Unlike swinepox virus whose infection in non-porcine species is non-productive, raccoonpox virus infection is productive, but may not spread systemically like vaccinia virus.
Raccoonpox virus was originally isolated from the upper respiratory tract of 2 of 92 apparently healthy raccoons in 1962 in Maryland (1). Significant HI (hemagglutinating-inhibiting) antibody titers for RPV were demonstrated in 23% [22/92] of raccoon sera indicating that RPV is indigenous in North America (2). Serological surveys testing sera from 593 raccoons in Delaware demonstrated that 4% were positive for RPV neutralizing antibodies, 12% were positive by ELISA and 2.5% of the sera were positive by both tests (10). RPV is classified as a orthopoxvirus based upon biological, serological and biophysical methods. RPV and volepox are the only orthopoxviruses thought to be native to North America. The Hind III DNA restriction map of RPV is the most divergent from other orthopoxvirus DNA maps, which are highly conserved, indicating that RPV has diverged from the other group members (4,6). Of 29 HindIII fragments from the RPV genome, none comigrated with HindIII fragments that were common to other examined orthopoxvirus DNA. RPV is naturally attenuated (10-100×) and avirulent compared with wild type vaccinia virus. There are four published RPV DNA sequences which include a Sal I end fragment (2.2 kb) (5), a hemagglutinin gene (1.5 kb) (7), a protein phosphatase sequence (0.5 kb; H1L ) (15), and the TK gene (834 bp) (16). RPV TK shows 87% amino acid homology to TK of vaccinia and 84.3% nucleotide homology. RPV HA shows 53% amino acid homology with vaccinia HA and 66% nucleotide homology. No other information regarding the genomic organization of RPV is known.
To date, recombinant raccoon poxviruses have been generated by insertion of foreign genes such as rabies virus glycoprotein into the TK locus of raccoon poxvirus (7-14). RPV recombinants expressing foreign viral antigens have been shown to elicit host protective immune responses in non-raccoon species including dogs, cats, and sheep with no negative effects observed.
The present invention provides a recombinant raccoon poxvirus comprising a foreign DNA inserted into the HindIII M region, HindIII N region or HindIII U region of the raccoonpox virus genome. The recombinant raccoon poxvirus is useful as a vaccine in mammalian and avian species. Prior to the present invention, it was unknown whether non-essential regions existed in the HindIII M, HindIII N, or HindIII U regions, and whether a stable recombinant raccoonpox virus could be isolated with foreign DNA insertions into these regions.
SUMMARY OF THE INVENTION
The present invention provides a recombinant raccoonpox virus comprising a raccoonpox virus viral genome which contains a foreign DNA sequence inserted into a non-essential region within the HindIII “U” genomic region of the raccoonpox region of the raccoonpox virus genome.
The present invention provides a recombinant swinepox virus comprising a swinepox virus genome which contains a foreign DNA sequence inserted into a non-essential region of the raccoonpox virus genome, wherein the foreign DNA sequence is a host range gene selected from the group consisting of raccoonpox virus K1L and raccoonpox virus C7L.
The present invention provides a recombinant raccoonpox virus comprising a raccoonpox virus viral genome which contains a foreign virus viral genome which contains a foreign DNA sequence inserted into a non-essential region within the HindIII “N” genomic region of the raccoonpox virus genome.
The present invention provides a recombinant raccoonpox virus comprising a raccoonpox virus viral genome which contains a foreign DNA sequence inserted into a non-essential region within the HindIII “M” genomic region of the raccoonpox virus genome.
The present invention provides a recombinant raccoonpox virus comprising a raccoonpox virus viral genome which contains a deletion in a raccoonpox virus host range gene of the viral genome, wherein the raccoonpox virus host range gene is selected from the group consisting of C7L, C6L, C5L, C4L, C3L, C2L, C1L, N1L, N2L, M1L, M2L, and K1L.
The present invention provides a homology vector for producing a recombinant raccoonpox virus by inserting a foreign DNA sequence into the raccoonpox virus genome which comprises a double-stranded DNA molecule consisting of double-stranded foreign DNA sequence encoding an antigenic polypeptide derived from an animal pathogen. Located at one end of the foreign DNA sequence, is double-stranded feline virus genomic DNA homologous to the genomic DNA located at one side of a non-essential site of the raccoonpox viral genomic DNA. Located at the other end of the foreign DNA sequence, is double-stranded raccoonpox virus genomic DNA homologous to the genomic DNA located at the other side of the same site.
REFERENCES:
patent: 5266313 (1993-11-01), Esposito et al.
patent: 5820869 (1998-10-01), Wasmoen et al.
patent: WO9640268 (1996-12-01), None
patent: WO9804684 (1998-02-01), None
Thomas, EK, et al., Further Characterization of Raccoonpox Virus. (1975)Arch. Virol.49(2-3): 217-227 (Exhibit E).
Graham KA, et al., The T1/35kDa family of poxvirus-secreted proteins bind chemokines and modulate leukocyte influx into virus-infected tissues. (Mar. 3, 1997)Virology,229(1):12-24 (Exhibit F).
Knight, JC, et al., Further Analyses of the Orthopoxviruses Volepox Virus and Raccoon Poxvirus (1992)Virology190:423-433 (Exhibit G).
PCT International Search Report issued in connection with Application No. PCT/US99/15565, International Filing date Sep. 7, 1999. (Exhibit H).
Cochran Mark D.
Junker David E.
Salimi Ali R.
Salkeld Pamela G.
Schering-Plough Veterinary Corp.
Zaradic Sandy S.
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