Drug – bio-affecting and body treating compositions – Whole live micro-organism – cell – or virus containing – Genetically modified micro-organism – cell – or virus
Reexamination Certificate
1999-10-19
2004-01-27
Wehbe′, Anne M. (Department: 1632)
Drug, bio-affecting and body treating compositions
Whole live micro-organism, cell, or virus containing
Genetically modified micro-organism, cell, or virus
C424S093100, C424S093210, C424S093400, C424S184100, C435S455000, C435S320100, C435S325000, C435S472000, C435S480000, C435S069100, C514S04400A
Reexamination Certificate
active
06682729
ABSTRACT:
The development of this invention was supported by the University of Maryland, Baltimore, Md.
FIELD OF THE INVENTION
The present invention relates to a method for introducing endogenous or foreign genes into animal cells using live invasive bacteria as vectors. The method allows for the delivery of eukaryotic expression cassettes encoding the endogenous or foreign genes into animal cells or animal tissue, and is useful for expressing, e.g., vaccine antigens, therapeutic agents, immunoregulatory agents, antisense RNAs, and catalytic RNAs, in animal cells or animal tissue.
BACKGROUND OF THE INVENTION
I. Live Bacterial Vector Vaccines
The advent of recombinant DNA technology has greatly accelerated the development of vaccines to control epidemic, endemic, and pandemic infectious diseases (Woodrow et al,
New Generation Vaccines: The Molecular Approach
, Eds., Marcel Dekker, Inc., New York, N.Y. (1989); Cryz,
Vaccines and Immunotherapy
, Ed., Pergamon Press, New York, N.Y. (1991); and Levine et al,
Ped. Ann
., 22:719-725 (1993)). In particular, this technology has enabled the growth of a new class of vaccines called bacterial vector vaccines (Curtiss, In:
New Generation Vaccines: The Molecular Approach
, Ed., Marcel Dekker, Inc., New York, N.Y., pages 161-188 and 269-288 (1989); and Mims et al, In:
Medical Microbiology
, Eds., Mosby-Year Book Europe Ltd., London (1993)). These vaccines can enter the host, either orally, intranasally or parenterally. Once gaining access to the host, the bacterial vector vaccines express an engineered prokaryotic expression cassette contained therein that encodes a foreign antigen(s) Foreign antigens can be any protein (or part of a protein) or combination thereof from a bacterial, viral, or parasitic pathogen that has vaccine properties (
New Generation Vaccines: The Molecular Approach
, supra;
Vaccines and Immunotherapy
, supra; Hilleman,
Dev. Biol. Stand
., 82:3-20 (1994); Formal et al,
Infect. Immun
. 34:746-751 (1981); Gonzalez et al,
J. Infect. Dis
., 169:927-931 (1994); Stevenson et al,
FEMS Lett
., 28:317-320 (1985); Aggarwal et al,
J. Exp. Med
., 172:1083-1090 (1990); Hone et al,
Microbial. Path
., 5:407-418 (1988); Flynn et al,
Mol. Microbiol
., 4:2111-2118 (1990); Walker et al,
Infect. Immun
., 60:4260-4268 (1992); Cardenas et al,
Vacc
., 11:126-135 (1993); Curtiss et al,
Dev. Biol. Stand
., 82:23-33 (1994); Simonet et al,
Infect. Immun
., 62:863-867 (1994); Charbit et al,
Vacc
., 11:1221-1228 (1993); Turner et al,
Infect. Immun
., 61:5374-5380 (1993); Schodel et al,
Infect. Immun
., 62:1669-1676 (1994); Schodel et al,
J. Immunol
., 145:4317-4321 (1990); Stabel et al,
Infect. Immun
., 59:2941-2947 (1991); Brown,
J. Infect. Dis
., 155:86-92 (1987); Doggett et al,
Infect. Immun
., 61:1859-1866 (1993); Brett et al,
Immunol
., 80:306-312 (1993); Yang et al,
J. Immunol
., 145:2281-2285 (1990); Gao et al,
Infect. Immun
., 60:3780-3789 (1992); and Chatfield et al,
Biol/Technology
, 10:888-892 (1992)). Delivery of the foreign antigen to the host tissue using bacterial vector vaccines results in host immune responses against the foreign antigen, which provide protection against the pathogen from which the foreign antigen originates (Mims,
The Pathogenesis of Infectious Disease
, Academic Press, London (1987); and
New Generation Vaccines: The Molecular Approach
, supra).
Of the bacterial vector vaccines, live oral Salmonella vector vaccines have been studied most extensively. There are numerous examples showing that Salmonella vectors are capable of eliciting humoral and cellular immunity against bacterial, viral and parasitic antigens (Formal et al,
Infect. Immun
., 34:746-751 (1981); Gonzalez et al, supra; Stevenson et al, supra; Aggarwal et al, supra; Hone et al, supra; Flynn et al, supra; Walker et al, supra; Cardenas et al, supra; Curtiss et al, supra; Simonet et al, supra; Charbit et al, supra; Turner et al, supra; Schodel et al, supra, Schodel et al (1990), supra; Stabel et al, supra; Brown, supra; Doggett et al, supra; Brett et al, supra; Yang et al, supra; Gao et al, supra; and Chatfield et al, supra). These humoral responses occur in the mucosal (Stevenson et al, supra; Cardenas et al, supra; Walker et al, supra; and Simonet et al, supra), and systemic compartments (Gonzalez et al, supra; Stevenson et al, supra; Aggarwal et al, supra; Hone et al, supra; Flynn et al, supra; Walker et al, supra; Cardenas et al, supra; Curtiss et al, supra; Simonet et al, supra; Charbit et al, supra; Turner et al, supra; Schodel et al, supra, Schodel et al (1990), supra; Stabel et al, supra; Brown, supra; Doggett et al, supra; Brett et al, supra; Yang et al, supra; Gao et al, supra; and Chatfield et al, supra). Live oral Salmonella vector vaccines also elicit T cell responses against foreign antigens (Wick et al,
Infect. Immun
., 62:4542-4548 (1994)). These include antigen-specific cytotoxic CD8
+
T cell responses (Gonzalez et al, supra; Aggarwal et al, supra; Flynn et al, supra; Turner et al, supra; and Gao et al, supra).
Ideally, bacterial vector vaccines are genetically defined, attenuated and well-tolerated by the recipient animal or human, and retain immunogenicity (Hone et al,
Vaccine
, 9:810-816 (1991); Tacket et al,
Infect. Immun
., 60:536-541 (1992); Hone et al,
J. Clin. Invest
., 90:412-420 (1992); Chatfield et al,
Vaccine
, 10:8-11 (1992); Tacket et al,
Vaccine
, 10:443-446 (1992); and Mims, supra). Recently, the number of potential bacterial vector vaccines for the delivery of prokaryotic expression cassettes has grown. They now include, but are not restricted to
Yersinia enterocolitica
(van Damme et al,
Gastroenterol
., 103:520-531 (1992)), Shigella spp. (Noriega et al,
Infect. Immun
., 62:5168-5172 (1994)),
Vibrio cholerae
(Levine et al, In:
Vibrio cholerae, Molecular to Global Perspectives
, Wachsmuth et al, Eds, ASM Press, Washington, D.C., pages 395-414 (1994)), Mycobacterium strain BCG (Lagranderie et al,
Vaccine
, 11:1283-1290 (1993); Flynn,
Cell. Molec. Biol
., 40(Suppl. 1):31-36 (1994)), and
Listeria monocytogens
(Schafer et al,
J. Immunol
., 149:53-59 (1992)) vector vaccines.
II. Eukaryotic and Prokaryotic Expression Cassettes
Normally, an expression cassette is composed of a promoter region, a transcriptional initiation site, a ribosome binding site (RBS), an open reading frame (orf) encoding a protein (or fragment thereof), with or without sites for RNA splicing (only in eukaryotes), a translational stop codon, a transcriptional terminator and post-transcriptional poly-adenosine processing sites (only in eukaryotes) (Wormington,
Curr. Opin. Cell Biol
., 5:950-954 (1993); Reznikoff et al,
Maximizing Gene Expression
, Eds., Butterworths, Stoneham, Mass. (1986)). The promoter region, the RBS, the splicing sites, the transcriptional terminator and post-transcriptional poly-adenosine processing sites are markedly different in eukaryotic expression cassettes than those found in prokaryotic expression cassettes (Wasylyk, In:
Maximizing Gene Expression
, supra, pages 79-99; Reznikoff et al, In:
Maximizing Gene Expression
, supra, pages 1-34; and Lewin,
Genes V
, Oxford University Press, Oxford (1994)). These differences prevent expression of prokaryotic expression cassettes in eukaryotic cells and visa versa (Miller et al,
Mol. Micro
., 4:881-893 (1990); and Williamson et al,
Appl. Env. Micro
., 60:771-776 (1994)).
Prokaryotic promoters are not active in eukaryotic cells and eukaryotic promoters are not active in prokaryotic cells (Eick et al,
Trends in Genetics
, 10:292-296 (1994)). The basis for the functional diversity of eukaryotic versus prokaryotic promoters is mediated by RNA-polymerase, transcription regulatory factors and the DNA structure of the promoter (Eick et al, supra).
RNA polymerases are DNA-binding proteins that recognize specific sequences in the DNA of promoter regions. RNA polymerases catalyze the synthesis of RNA molecules by polymerizing nucleoside triphosphates in the specific order that is dictated by the DNA coding sequence (Libby et al,
Mol. Micro
., 5:
Hone David M.
Lewis George K.
Powell Robert J.
University of Maryland Baltimore
Wehbe′ Anne M.
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