Method for introducing and expressing RNA in animal cells

Details Method for introducing and expressing RNA in animal cells Method for introducing and expressing RNA in animal cells

2000-04-07

2002-12-31

Priebe, Scott D. (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, C435S252300, C435S320100, C435S455000, C514S04400A

Reexamination Certificate

active

06500419

ABSTRACT:

BACKGROUND OF THE INVENTION
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 nucleic acid vaccines. Although the first nucleic acid vaccine was only reported in 1992, the study of nucleic acid vaccines has grown dramatically and examples of this approach have been reported in a wide array of animals using numerous antigens (Tang, D. C., et al. (1992)
Nature
356:152; Fynan, E. F. et al. (1993)
PNAS USA
90:11478; Donnelly, J. J. et al. (1995)
Nat Med
1:583; Wang, B. et al. (1993)
PNAS USA
90:4156; Davis, H. L., et al. (1993)
Hum Mol Genet
2:1847; Ulmer, J. B. et al. (1993)
Science
259:1745; Robinson, H. L. et al. (1993)
Vaccine
11:957; Eisenbraun, M. D. et al. (1993)
DNA Cell Biol
12:791; Wang, B. et al. (1994)
AIDS Res Hum Retroviruses
10:S35; Coney, L. et al. (1994)
Vaccine
12:1545; Sedegah, M. et al. (1994)
Proc Natl Acad Sci USA
91:9866; Raz, E. et al. (1994)
Proc Natl Acad Sci USA
91:9519; Xiang, Z. Q. et al. (1994)
Virology
199:132) (for a more comprehensive list see www.genweb.com/ Dnavax/Biblio/articles). Early murine experiments with HIV-1 DNA vaccines produced impressive immunological responses, including neutralizing antibody responses against HIV-1 and strong CTL responses against several HIV-1 antigens (Wang, B. et al. (1993) supra; Wang, B. et al. (1994) supra; Coney, L. et al. (1994) supra; Lu, S. et al. (1995)
Virology
209:147; Shiver, J. W. et al. (1995)
Annals of the New York Academy of Sciences
772:198; Wahren, B. et al. (1995)
Ann N Y Acad Sci
772:278). However, an initial attempt to induce protective immunity with an SIV DNA vaccine in Rhesus monkeys was disappointing (Lu, S. et al. (1996)
J Virol
70:3978). In contrast, an HIV-1
MN
Env DNA vaccine induced measurable protection against a chimeric SIV/HIV (SHIV) challenge in vaccinated cynomologous macaques (Boyer, J. D. et al. (1996)
Journal of Medical Primatology
25:242). More recently, intramuscular immunization of chimpanzees with this latter vaccine engendered protection against parenteral challenge with HIV-1
SF-2
(Boyer, J. D. et al. (1997)
Nature Medicine
3:526). These differences may have resulted from antigenic differences in the vaccines or differences in the potency of the challenges (Lu, S. et al. (1996) supra; Boyer, J. D. et al. (1996) supra; Boyer, J. D. et al. (1997) supra). It is noteworthy to mention that in the aforementioned chimpanzee study, DNA vaccination only induced modest humoral and cellular responses despite giving 9 doses of vaccine containing a total of 2.9 mg of DNA before challenge (Boyer, J. D. et al. (1997) supra). Thus, although these results are encouraging, the immunogenicity of HIV-1 DNA vaccines must be improved before this approach achieves practical utility in large scale vaccination programs.
The mechanism though which DNA vaccines induce immunity is not fully understood. Muscle cells express low levels of MHC class 1 and do not express detectable levels of co-stimulatory molecules B7-1 and B7-2 (review by Ertl, H. C. and Z. Q. Xiang (1996)
Journal of Immunology
156:3579). While it remains conceivable that muscle cells may serve as an antigen depot (Ertl and Xiang (1996) supra), their participation in the induction of MHC class I and II responses may be secondary to other antigen presenting cells (Ertl and Xiang (1996) supra). Xiang and Ertl (Ertl and Xiang (1996) supra; Xiang, Z. and H. C. Ertl (1995)
Immunity
2:129) have suggested that resident dendritic cells may be involved in the primary inductive events. They showed that co-expression of GM-CSF, a cytokine know to activate growth of dendritic cells, at the site of inoculation resulted in a more rapid response to DNA vaccine encoded antigens (Xiang and Ertl (1995) supra). In contrast, co-expression of IFN-&ggr; diminished the responses (Xiang and Ertl (1995) supra). In agreement, Manickan et al. ((1997)
Journal of Leukocyte Biology
61:125) showed that immunization with dendritic cells transfected with a DNA vaccine induced elevated immune responses, compared to the identical DNA vaccine given alone. In addition, dendritic cells have been shown to express antigen following intradermal vaccination with a DNA vaccines (Raz, E. et al. (1994) supra). Although inconclusive, these data strongly suggest that dendritic cells may play a substantial role in the presentation of DNA vaccine-encode antigens.
Another new class of vaccines are 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,
Bio/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 a

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