Chemistry: molecular biology and microbiology – Virus or bacteriophage – except for viral vector or...
Reexamination Certificate
2000-03-24
2003-09-02
Smith, Lynette R. F. (Department: 1645)
Chemistry: molecular biology and microbiology
Virus or bacteriophage, except for viral vector or...
C435S091330, C424S204100, C424S218100
Reexamination Certificate
active
06613556
ABSTRACT:
FIELD OF THE INVENTION
This invention relates to virus replicated in vertebrate cell cultures, which are suitable for use in vaccine production. Viral strains useful as vaccines can be adapted to vertebrate cells by serial passage and optimization of replication of the vaccine strains. Vaccine strains adapted for growth in vertebrate cells offer many advantages. The invention also relates to methods for making and using such replicated viruses such as for vaccine.
BACKGROUND OF THE INVENTION
Cultured vertebrate cells have been used for virus replication and have been classified into at least three distinct groups. Primary cells, derived from normal tissue; diploid cell strains are those that have a limited, finite life span, depending on the species of origin; and continuous cell lines, cultured cells that replicate indefinitely and may be capable of growth in suspension culture. Hayflick, in
Continuous Cell Lines as Substrates for Biologicals
, Arlington, Va., p2 (1988).
At present most viral vaccines are produced using primary cells or diploid cell strains. Species of origin include monkeys, chicken, embryos, and hamsters. These cell cultures have certain advantages such as easy preparation using simple media and bovine sera and sensitivity to a wide range of viruses. However, primary cells suffer from disadvantages, such as contamination by various adventitious agents, variable quality and sensitivity, and difficulty in obtaining suitable tissue for cell cultivation, low virus titers and the high cost of obtaining and preparing such cell cultures. Diploid cell strains have many of the same disadvantages as well as a finite life span that limits expansion of the cells.
African green monkey kidney (Vero) cells are a continuous cell line, that is not tumorigenic, and is suitable and advantageous for human vaccine production. In fact, several licensed vaccines are currently manufactured in these cells. Vero cells, and possibly other continuous cell lines, may be suitable for dengue virus vaccine production. However, it is not known whether the antigenic characteristic of a vaccine virus strain would not be altered after passaging in a continuous cell line.
SUMMARY OF THE INVENTION
The present invention relates to methods, replicated viruses and vaccine compositions using high growth strains of dengue virus.
The invention thus provides for a method for increasing dengue viral replication in culture by passaging the desired viral strain in continuous vertebrate cells and choosing the viral strain which replicates to high titer as the seed strain. When the dengue strain is attenuated, the resulting seed strain can be suitable for production of vaccine.
Preferably, the vertebrate host cells are continuous cell cultures derived from epithelial cells or fibroblasts, as mammalian cell lines of passage number 10-250. Preferably used for vaccine production are Vero cells as a continuous line of a passage number of about 20-250, currently available and certified (e.g., by the WHO).
Dengue virus of the invention, in isolated, purified or concentrated from, preferably has an infectivity titer of about 10
6
-10
9
(such as 10
6
-10
7
, 10
7
and 10
8
-10
9
, or any range or value therein) plaque forming units (PFU) per ml.
The present invention also provides vaccine compositions comprising at least one strain of a dengue virus of any of the four serotypes of dengue of the present invention, in inactivated or attenuated form, optionally further comprising at least one of: (a) at least one pharmaceutically acceptable carrier or diluent; (b) at least one adjuvant and/or (c) at least one therapeutic agent.
The present invention also provides a method for eliciting an immune response to at least one dengue virus strain in a mammal, which response is prophylactic or therapeutic for a dengue virus infection. The method comprises administering to the mammal a vaccine composition comprising an inactivated and/or attenuated dengue virus of the present invention. The composition is provided in an amount that is protective or therapeutic for the mammal against a dengue virus pathology caused by infection with dengue virus.
Other objects, features, advantages, utilities and embodiments of the present invention will be apparent to skilled practitioners from the following detailed description and examples relating to the present invention.
REFERENCES:
Smith et al. 1985 J. Gen. Virol. pp. 559-571.*
Kraiselburd et al 1987 Publication GRAI8723.*
Osatomi, K., “Complete Nucleotide Sequence of Dengue Type 3 Virus Genome RNA”, Virology 176:643-647 (1990).
Puri, B., “Molecular analysis of dengue virus attenuation after serial passage in primary dog kidney cells”, J. General Virology (1997) 78:2287-2291.
Botstein and Shortle, 1985, “Strategies and applications of in vitro mutageNesis”, Science, vol. 229, No. 4719, pp. 1193-1201.
Clarke and Casals, 1958, “Techniques for hemagglutination and hemagglutination-inhibition with arthropod-borne viruses”, Am. J. Trop. Med. Hyg., 7, 561-573.
Halstead et al., 1984, “Selection of attenuated Dengue 4 viruses by serial passage in primary kidney cells,” Am. J. Trop. Med. Hyg. 33(4), pp. 654-665.
Halstead et al., 1984, “Selection of attenuated Dengue 4 viruses by serial passage in primary kidney cells,” Am J. Trop. Med. Hyg, 33(4), pp. 666-671.
Halstead et al., 1984, “Selection of attenuated Dengue 4 viruses by serial passage in primary kidney cells,” Am J. Top. Med. Hyg, 33(4), pp. 672-678.
Halstead et al., 1984, “Selection of attenuated Dengue 4 viruses by serial passage in primary kidney cells,” Am J. Trop. Med. Hyg, 33(4), pp. 679-683.
Hayflick, 1988, “History of cell substrates used for human biologicals”, Symposium on Continuous Cell Lines as Substrates for Biologicals, Arlington, Virginia, USA, pp. 11-26.
Hoke et al., 1990, “Preparation of an attenuated Dengue 4 virus vaccine”, Am J. Top. Med., Hyg., 43(2), pp. 219-226.
Marchette, 1990, “Preparation of an attenuated Dengue 4 virus vaccine”, Am J. Top. Med. Hyg., 43(2), pp. 212-218.
Mizrahi, ed., Viral Vaccines, “WHO Attitude to Viral Vaccines”, Wiley-Liss, New York (1990), pp. 39-60.
Putnak et al, 1996, “Development of a purified inactivated, Dengue-2 virus, vaccine prototype in vero cells: immunogenecity and protection in mice and Rhesus monkeys,” J. Infectious Dis., 174, pp. 1176-1184.
Russell, et al., “A plaque reduction test for dengue virus neutralizing antibodies”, J. Immunology, vol. 99, No. 2, 1967, pp. 285-290.
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Sukhavachana et al, 1966, “Tissue culture techniques for the study of dengue viruses”, Abreges des Communications, Bull. WHO 35, pp. 65-66.
Zollerf and Smith, 1984, Laboratory Methods, “Oligonucleotide-directed mutagenesis: a simple method using two oligonucleotide primers and a single-stranded DNA template”, DNA, vol. 3, No. 6, pp. 479-488.
Bhamarapravati, 1987, “Immunization with a live attenuated dengue-2-virus candidate vaccine: clinical, immunolgical and biological responses in adult volunteers”, Bull. WHO, 65(2), pp. 189-195.
Conrad et al., “Infection with Nippostrongylus Brasiliensis or injection of anti-IgD antibodies markedly enhances Fc-receptor-mediated interleukin 4 produciton by non-B, non-T Cell,”, J. Exp. Med., vol. 171, pp. 1497-1508.
Dharakul, et al., “Dengue virus-specific memory T cell responses in human volunteers receiveing a live attenuated dengue virus Type 2 candidate vaccine”, J. Infect. Dis., vol. 170, pp. 27-33.
Edelman et al., “A live attenuated Dengue-1 vaccine candidate passaged inprimary dog kidney cell cultures is attenuated and immunogenic for humans”, 1994, Am. J. Trop. Med. Hyg., 170, pp. 1448-1455.
Halstead, 1978, “Studies on the attenuation of Dengue 4”, Asian J. Infectious Dis., vol. 2, pp. 112-117.
Halstead, 1970, “Long ‘cure ’improves results of pig heterograft heart valves,” JAMA, vol. 211, No. 6, pp. 911-916.
Johnson and Roehrig, “New mouse model for Dengue virus vaccien testing”, J. Virology, Jan. 1999,
Eckels Kenneth H.
Innis Bruce L.
Putnak Joseph R.
Arwine Elizabeth
Smith Lynette R. F.
The United States of America as represented by the Secretary of
Zeman Robert A.
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