Chemistry: molecular biology and microbiology – Process of mutation – cell fusion – or genetic modification – Introduction of a polynucleotide molecule into or...
Reexamination Certificate
1999-07-09
2004-08-03
Paras, Peter (Department: 1632)
Chemistry: molecular biology and microbiology
Process of mutation, cell fusion, or genetic modification
Introduction of a polynucleotide molecule into or...
C435S455000, C435S320100, C435S325000, C514S04400A, C424S093100, C424S093200, C424S093600
Reexamination Certificate
active
06770479
ABSTRACT:
FIELD OF THE INVENTION
This invention relates to vaccines for bacterial toxins from
Bacillus anthracis.
INTRODUCTION
Anthrax is a disease cause by the sporulating bacteria
Bacillus anthracis
. Humans working with animal products are at risk from contracting anthrax. Areas such as Iran, Turkey, Iraq, Pakistan, and sub-Saharan Africa are hyperendemic for anthrax, although the organism can be found in most areas of the world. Anthrax manifests disease in three different ways. Inhalation, gastrointestinal, and cutaneous anthrax result from inhaling spores, ingesting spores in contaminated meat, or contacting spores in an open wound, respectively. Untreated inhalation or gastrointestinal anthrax has a case fatality rate of essentially 100 percent while cutaneous anthrax has a case fatality rate of up to 25 percent. Previous research has shown that the protective antigen (PA) produced by
B. anthracis
can protect mice from anthrax. Even though the Anthrax vaccine is FDA licensed, reactogenicity is mild to moderate.
Therefore, there is a need for an efficacious vaccine for anthrax useful for protecting humans.
SUMMARY OF THE INVENTION
The present invention satisfies the need discussed above. The present invention relates to a method and composition for use in inducing an immune response which is protective against infection with anthrax.
In this vaccine strategy, a gene coding for a protein of interest is cloned into a VEE virus vector in place of the VEE virus structural genes; the result is a self-replicating RNA molecule, a replicon, that encodes its own replicase and transcriptase functions, and in addition makes abundant quantities of mRNA encoding the foreign protein. When replicon RNA is transfected into eukaryotic cells along with two helper RNAs that express the VEE structural proteins (glycoproteins and nucleocapsid), the replicon RNA is packaged into VEE virus-like particles by the VEE virus structural proteins, which are provided in trans. Since the helper RNAs lack packaging signals neccessary for further propagation, the resulting VEE replicon particles (VRPs) which are produced are infectious for one cycle but are defective thereafter. Upon infection of an individual cell with a VRP, an abortive infection occurs in which the infected cell produces the protein of interest in abundance, is ultimately killed by the infection, but does not produce any viral progeny (Pushko et al., 1997
, Virology
239, 389-401).
The PA gene which contains a prokaryotic secretory signal and the entire 83 kDa coding sequence (Welkos et al., 1988
, Gene
69, 287-300), was inserted into the VEE replicon vaccine vector (
FIG. 1
) and have demonstrated high level expression of this bacterial protein in eukaryotic cells in culture. Mice, either the C57BL/6 strain or the A/J strain, inoculated with VRP containing the PA-replicon produced high specific antibody titers and were protected from developing anthrax when challenged subcutaneously with
B. anthracis.
Therefore, it is one object of the present invention to provide a VEE virus replicon vector comprising a VEE virus replicon and DNA fragments encoding the PA protein from
B. anthracis.
It is another object of the present invention to provide a self replicating RNA comprising the VEE virus replicon and any of the
B. anthracis
fragments described above.
It is another object of the present invention to provide infectious VEE virus replicon particles produced from the VEE virus replicon RNA described above.
It is further an object of the invention to provide an immunological composition for the protection of mammals against
B. anthracis
infection comprising VEE virus replicon particles containing any of the
B. anthracis
fragments described above or a combination of different VEE virus replicons each having a different
B. anthracis
fragment.
REFERENCES:
patent: WO 95/07994 (1995-03-01), None
patent: WO 96/17067 (1996-06-01), None
patent: WO 96/37616 (1996-11-01), None
patent: WO 98/08952 (1998-03-01), None
SH Orkin et al., “Report and Recommendations of the Panel to Assess the NIH Investment in Research on Gene Therapy,” Dec. 1995, pp. 1-41.*
M Chattergoon et al., FASEB, “Genetic immunization: a new era in vaccines and immune therapeutics,” May 11 1997, pp. 753-763.*
WM McDonnell et al., The New England Journal of Medicine, “Molecular Medicine DNA Vaccines,” Jan. 1996,vol. 334, No. 1, pp. 42-45.*
FD Ledley, Human Gene Therapy, “Clinical Considerstions in the Design of Protocols for Somatic Gene Therapy,” 1991, 2:77-83.*
RM Zinkernagel, Fundamental Immunology, “Immunity to Viruses,” Chap. 34, 3rdEd., pp. 1211-1250.*
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Iocono-Connors et al., “Protection against Anthrax with Recombinant Virus-Expressed Protective Antigen in Experimental Animals”, Infection and Immunity, Jun. 1991, pp. 1961-1965.
Pushko et al., “Replican-Helper Systems from attenuated Venezuelan Equine Encephalitis Virus Expression of Heterologous Genes in Vitro and Immunization against Heterologous Pathogens In Vivo”, Virology, 239, pp. 389-401 (1997).
Singh, et al., “A Deleted Variant ofBacillus antracixProtective Antigen is Non-toxic and Blocks Anthrax Toxin Action in Vivo”, The Journal of Biological Chemistry, vol. 264, No. 32, pp. 19101-19107, Nov. 15, 1989.
Ivins and Welkos, “Cloning and Expression of the Bacillus anthracis Protective Antigen Gene inBacillus subtills”, Infectin and Immunity, vol. 54, No. 2, pp. 537-542 (Nov. 1986).
Clayton et al., “Protective Vaccination with a Recombinant Fragment of Clostridium botulinum Neurotoxin Serotype A Expressed frm a Synthetic Gene inE. coli”, Infection and Immunity, Jul. 1995, vol. 63, No. 7, pp. 2738-2742.
Byrne, et al., “Purification, Potency, and Efficacy of the Botulinum Neurotoxin Type A Binding Domain fromPichia pastoris as a Recombinant Vaccine Candidate”, Infection and Immunity, Oct. 1998, vol. 66, No. 10, pp. 4817-4822.
Bavari, et al., “Engineered Bacterial Superantigen Vaccines”, Vaccines 96, 1996, pp. 135-141.
Welkos et al., “Sequence and analysis of the DNA encoding protective antigen ofBacillus anthracis”, Gene, 69, 1988, pp. 287-300.
Lee John S.
Parker Michael D.
Pushko Peter
Smith Jonathan F.
Welkos Susan L.
Arwine Elizabeth
Harris Charles H.
Paras Peter
The United States of America as represented by the Secretary of
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