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WO1997012981A1 - Adenovirus recombine de la rage - Google Patents

Adenovirus recombine de la rage Download PDF

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Publication number
WO1997012981A1
WO1997012981A1 PCT/CA1996/000665 CA9600665W WO9712981A1 WO 1997012981 A1 WO1997012981 A1 WO 1997012981A1 CA 9600665 W CA9600665 W CA 9600665W WO 9712981 A1 WO9712981 A1 WO 9712981A1
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WO
WIPO (PCT)
Prior art keywords
adenovirus
mammal
rabies
vaccine
immune response
Prior art date
Application number
PCT/CA1996/000665
Other languages
English (en)
Inventor
Oksana K. Yarosh
Ludvik Prevec
Frank L. Graham
Original Assignee
Microbix Biosystems Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority to AU70818/96A priority Critical patent/AU7081896A/en
Application filed by Microbix Biosystems Inc. filed Critical Microbix Biosystems Inc.
Priority to EP96931711A priority patent/EP0859846A1/fr
Publication of WO1997012981A1 publication Critical patent/WO1997012981A1/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/86—Viral vectors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
    • C12N2710/00011—Details
    • C12N2710/10011—Adenoviridae
    • C12N2710/10311—Mastadenovirus, e.g. human or simian adenoviruses
    • C12N2710/10341—Use of virus, viral particle or viral elements as a vector
    • C12N2710/10343—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/20011—Rhabdoviridae
    • C12N2760/20111—Lyssavirus, e.g. rabies virus
    • C12N2760/20122—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

Definitions

  • the present invention relates to vaccines and more particularly to vaccines against rabies virus.
  • the rhabdovirus rabies is known to be spreading among selected wild animal species in Eastern Central North America. According to figures released in an August 1993 issue of the New York Times, 1098 people were treated for rabies in New York State in 1992 compared to only 81 in 1989.
  • Vaccination continues to be the only effective way to prevent or to treat rabies and other rhabdovirus infections.
  • the objective of vaccinating wildlife species against rabies is to reduce the number of non-protected animals to the point where the disease cannot sustain itself in nature.
  • Rabies vaccines consisting of killed or highly attenuated rabies virus, have been available since the time of Rasteur. Attenuated rabies strains currently used to vaccinate North American wildlife fail to effectively immunize several vector species and are pathogenic in rodents. Attempts have been made to overcome these problems with limited success.
  • the invention involves a recombinant adenovirus adapted to raise in a mammal an immune response to rabies, comprising a functionally disrupted El region and an expression cassette in which DNA coding for rabies glycoprotein is linked operably with expression controls functional in a eukaryotic host.
  • a vaccine for rabies virus to be administered to mammals comprising recombinant adenovirus having a functionally disrupted El region and an expression cassette in which DNA coding for rabies glycoprotein is linked operably with expression controls functional in a eukaryotic host in an amount effective to generate an immune response in the recipient mammal, and a vehicle suitable for delivering the adenovirus to the mammal.
  • a method useful to immunize a mammal against rabies comprising the step of delivering a dosage of an adenovirus sufficient to generate an immune response to rabies in the mammal, the adenovirus comprising a functionally disrupted El region and an expression cassette in which DNA coding for rabies glycoprotein is linked operably with expression controls functional in a eukaryotic host.
  • a method of producing a defective adenovirus vaccine comprising the steps of: culturing in an El producing eukaryotic host, an adenovirus having a functionally disrupted El region and an expression cassette in which DNA coding for rabies glycoprotein is linked operably with expression controls functional in the eukaryotic host, and harvesting the replicated adenovirus.
  • Figure 1 is a schematic representation of recombinant adenovirus construct
  • Figure 2 is a schematic representation of certain plasmids involved in the preparation of the construct illustrated in Figure 1 ;
  • Figure 3 is an autoradiogram of immuno-precipitated protein expressed by the construct of Figure 1.
  • the invention concerns a recombinant adenovirus of the type having a linear double stranded genome ranging in size from 30 to 40 kbp.
  • the virion is a non-enveloped icosahedron, approximately 70 nm in diameter composed of proteins and DNA.
  • the double stranded genome has what is known as a right strand and a left strand.
  • the adenovirus is known to include early regions El and E3 in the right strand.
  • a particular feature of the present recombinant adenovirus is the presence of an a functionally disrupted El region and an expression cassette in which DNA coding for rabies glycoprotein is linked operably with expression controls functional in a eukaryotic host.
  • 'Eukaryotic hosts' include adenovirus hosts including mammalian cells including cell lines such as 293, MRC-5, MDCK and HeLa and most preferably El producing hosts such as 293 cells.
  • 'Expression controls' are DNA sequences that serve to drive the transcription of the rabies glycoprotein-encoding DNA and include such functional elements as a promoter and a terminator.
  • the promoter is from cytomegalovirus (CMV) and more preferably from a human cytomegalovirus (hCMV) and most desirably an hCMV immediate early (IE) promoter. While hCMV is preferable, other promoters may be used including, for example, murine cytomegalovirus immediate early promoter or adenovirus type 2 major late promoter. Suitable terminators are well known and include SV40 polyadenylation sequence and termination region.
  • An 'immune response' is considered 'raised' when neutralizing antibodies can be detected in serum taken within about three to four months post immunization from a recipient mammal using such detection techniques as an ELISA assay, or other suitable methods of detection such as a rabies fluorescence inhibition microtest (FIMT) described by Zalan et al., (1979 Biological Standards, vol. 7, pp 213-220).
  • FIMT rabies fluorescence inhibition microtest
  • the expression cassette is incorporated within a cloning site available in the functionally disrupted El region of the adenovirus and more desirably, the early region El is deleted and the expression cassette is substituted therefor.
  • the rabies glycoprotein-encoding DNA or the expression cassette may be inserted, so long as the insertion does not interfere with the ability of the recombinant virus to replicate in suitable cells.
  • the rabies glycoprotein-encoding DNA encodes for rabies glycoprotein G, and most desirably for rabies glycoprotein G derived from ERA strain of rabies, though rabies glycoprotein-encoding DNA from other strains may also be used including those derived from arctic fox rabies variants.
  • the expression cassette may be formed by a number of plasmid vector routes.
  • the rabies glycoprotein-encoding DNA is obtained from plasmid pSV2RGla and the recombinant adenovirus is formed by ligating the rabies glycoprotein-encoding DNA into a suitable cloning site of plasmid pHMCVspl3 and co-transfecting the plasmid pHMCVspl3 with plasmid pJM17 into 293 cells.
  • glycoprotein-encoding DNA and accompanying SV40 polyadenylation and termination sequences may be isolated by ligating the approximately 1.8 kb fragment obtained from plasmid pSV2RGla after complete digestion with Xbal, to form a rabies glycoprotein- encoding DNA fragment which may be gel purified and ligated into the Xbal site of pHCMVspl3 in an orientation parallel to its HCMV IE promoter.
  • the resultant plasmid pDCHCMV13RG may then be co-transfected with pJM17 into 293 cells according to the protocol described by Graham and Prevec (Mol. Biotechnology vol.
  • the rabies glycoprotein-encoding DNA sequences may be inserted into any one of pCA3, pCA4, pCA13 or pCA14 (available from MICROBIX BIOSYSTEMS INC.) using one or more of the available cloning sites between the HCMV promoter in each and polyadenylation sequences in each of these plasmids to form a shuttle plasmid.
  • the shuttle plasmid can then be combined with one of pJM17, pBHGlO pBHGll , or pBHGE3 by co- transfection of 293 cells using the calcium phosphate technique (Graham and van der Eb (1973) Virology 52:456-467) to produce a recombinant Ad5HMCV13RG vector.
  • the resulting vectors from the second route have a deleted El region and therefore are defective, some have a disrupted E3 region, including a deleted or substituted E3 region structure.
  • the resulting vector when formed from pJM17, the resulting vector has an E3 region essentially identical to that produced by the first route.
  • the resulting vector has a deleted E3 region while with pBHGE3, the resulting vector has an intact E3 region.
  • a vaccine for rabies virus to be administered to mammals comprising an amount of recombinant adenovirus having a functionally disrupted El region and an expression cassette in which DNA coding for rabies glycoprotein is linked operably with expression controls functional in a eukaryotic host, and a vehicle suitable for delivering said adenovirus in the mammal.
  • a method useful to immunize a mammal against rabies comprising the step of delivering a dosage of an adenovirus sufficient to generate an immune response to rabies in the mammal, said adenovirus comprising a functionally disrupted El region and an expression cassette in which DNA coding for rabies glycoprotein is linked operably with expression controls functional in a eukaryotic host.
  • the resulting defective recombinant adenovirus is cultured by infecting El -producing eukaryotic host cells such as 293 cells available from MICROBIX BIOSYSTEMS INC (Graham et al, (1977) J.
  • the aim of the vaccine is to deliver to the mammal, an amount of the recombinant adenovirus effective to generate an immune response in the mammal to rabies virus.
  • the vaccine may be effective at a strength of 10 6 or more plaque forming units (pfu). For instance, a strength of about 10 8 pfu is suitable for use in vaccinating skunks.
  • the dosage will of course otherwise vary with the animal treated and will also vary according to how the vaccine is administered and may also be effective at lower dosages.
  • the vaccine may be injected intramuscularly, intraperitoneally, or intravenously with a suitable vehicle such as phosphate buffered saline (PBS) solution.
  • a suitable vehicle such as phosphate buffered saline (PBS) solution.
  • the recombinant adenovirus may be administered orally by direct oral installation or by way of an edible bait material with sufficient virus in a single edible bait material, to generate a immune response in wild animals such as skunks or raccoons.
  • Example 1 HAd5 El RABIES GLYCOPROTEIN GENE RECOMBINANT As will be described with reference to Figures 1 and 2, the vector
  • Ad5HCMB13RG contains the rabies glycoprotein gene driven by the human cytomegalovirus (HCMV) immediate early (IE) promoter inserted into an El region deletion.
  • the expression cassette is constructed with the rabies glycoprotein-encoding DNA flanked upstream by the HCMV immediate early promoter and flanked downstream by the SV40 polyadenylation and terminator sequences.
  • the expression cassette containing the rabies glycoprotein- encoding DNA replaces El region sequences from 0.9 to 9.8 map units (mu) in the adenovirus genome.
  • the vector Ad5HCMB13RG is derived from the following:
  • pJM17 pJM17 (available from MICROBIX BIOSYSTEMS INC.) is a derivative of pFG140 (available from MICROBIX BIOSYSTEMS INC.), which is itself derived from the Ad5 mutant dll309 (Jones and Shenk, (1978) Cell 13:181-188; (1979) Cell 17:683-689).
  • Ad5 mutant dll309 Jones and Shenk, (1978) Cell 13:181-188; (1979) Cell 17:683-689.
  • the detailed differences between dl309 and wild type Ad5 in die E3 region have been described by Bett et al.
  • pSV2X3 pSV2X3 contains the SV40 early promoter and polyadenylation sequences separated by a multi-cloning site (HindlH BamHI EcoRI Smal Xhol) (Prevec et al., (1990) J. Infect. Disease 161:27-30)
  • PSV2RG1 pSV2RGl (Prevec et al., (1990) J. Infect. Disease 161:27-30) is derived from pSV2X3 and contains an approximately 1.7 kb EcoRI to BamHI fragment of rabies (ERA strain) glycoprotein G cDNA cloned into the unique Smal site and oriented parallel to the SV40 promoter.
  • pSV2RGla was cut with Ncol and a self-annealed
  • Xbal linker AB1185 (5'CATGTCTAGA 3' ) was ligated into this site. E. coli cells were transformed with the ligation mix and plasmid cDNA was screened for an additional Xbal site replacing a 120 bp Ncol fragment to form pSV2RGla, the modifications to which were confirmed by sequencing with primer AB 1180.
  • pHCMVsp!3 pHCMVs ⁇ l3 is a derivative of the generic pBR322 plasmid and contains left-end sequences of Ad5 (nt- 1-5788) with an Aflll/SlpI El region deletion (nt 342-3533). The deletion was replaced with the SlpI binding site for protein IX, the human cytomegalovirus (HCMV) immediate early (IE) promoter, and a poly-linker region. The hCMV IE promoter and poly-linker sequences were oriented so that transcription from the hCMV promoter of genes inserted into the poly-linker site is toward the left end of the resultant adenovirus vector.
  • HCMV human cytomegalovirus
  • IE immediate early
  • Ad5HCMV13RQ Rabies glycoprotein and SV40 poly A sequences were first isolated by cutting pSV2RGla with Xbal to form the 1.8 kb Xbal fragment, which was later gel-purified and ligated into the Xbal site of pHCMVspl3 in an orientation parallel to the HCMV IE promoter.
  • the resultant plasmid pDCHCMV RG was co-transfected with pJM17 into 293 cells according to the protocol developed by Graham and van der Eb ((1973) Virology 52:456-467) as described by Graham and Prevec (Mol. Biotechnology vol. 3, 207-220 (1995)).
  • IX citrate-saline Large 150mm dishes of 293 cells were washed with IX citrate-saline, and set up into 60mm dishes for 70-80% confluence at the time of use, usually the following day. Five dishes were prepared per DNA concentration per plasmid.
  • IX HEPES buffered saline 20 mM HEPES (nN-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid), 137 mM NaCl, 5 mM KC1, 0.7 mM Na 2 HPO 4 2H 2 O, 6mM glucose, final pH 7.1, autoclaved and stored at 4 degrees Celsius
  • the resultant Ad5HCMV13RG carries the rabies glycoprotein gene flanked by die HCMV IE promoter and the SV40 polyA addition sequences within a deletion of the El region of an Ad5 virus derived from pJM17 and with modified E3 region as described above under pJM17.
  • Rabies glycoprotein expression by Ad5HMCV13RG was examined in cell lines that were either able or unable to provide El helper function in trans. El region coding sequences are required in trans for Ad5HMCV13RG replication as the virus contains a deletion widiin El.
  • Cell lines 293, MRC-5, MDCK and HeLa were infected widi either Wild Type hAd5 or with recombinant Ad5HMCV13RG at a multiplicity of infection (MOI) of 100 pfu/cell.
  • MOI multiplicity of infection
  • the Wild Type hAd5 was strain dl309, which contains an E3 deletion which removes both the EcoRI site (83.6 mu) and Xbal site (84.8 mu) and a base substitution at the 78.3 mu Xbal site (Jones and Shenk, (1979) Cell 17:683-689). Infected cells were metabolically labelled with [ 35 S] methionine prior to harvesting at either 9 or 25 hours post immunization. Rabies glycoprotein was immuno-precipitated with 3 ⁇ l of ascites-derived monoclonal antibodies. Although an El deleted vector of this type can infect all these cell types, it is unable to replicate in all cell lines except for 293. Any detectable rabies glycoprotein expression occurs as a result of the hCMV promoter and, in the absence of replication, the levels of glycoprotein expression depend on the MOL
  • rabies glycoprotein expression was detected in human 293 cells or HeLa cells or MRC-5 cells and in canine MDCK cells. As expected the highest levels of expression were obtained with 293 cells and diese levels were approximately 10 to 100 fold higher than levels in cells non- permissive for virus replication. Cells, other than 293, infected with Ad5HCMV13RG, failed to exhibit a cytopathic effect for an extended period of time as compared to wild type virus, and would therefore be expected to continue to produce the glycoprotein for extended periods.
  • Vaccine was prepared in the following manner and according to the technique described by Hitt et al. (1994 Cell Biology: A laboratory handbook, Academic Press pp 479-490). 150mm dishes of 293 cells were grown to be 80 to 90% confluent at me time of infection. The medium was removed from the
  • Ad5HCMV13RG 293 cells and the cells were infected with Ad5HCMV13RG at a multiplicity of infection of 1-10 PFU per cell (1 ml virus suspension per 150mm dish).
  • Adenovirus titres were determined by assaying for viral plaques on 293 cells. Cells were set up in 60mm dishes one day prior to use so as to have the cells at about 90% confluence at the time of infection. Generally, eight 60mm dishes of 293 cells were seeded from each 150mm dish. Virus dilutions (1 : 10 series) were prepared in PBS+ -f- . Medium was removed, die cells were washed once witii PBS+ + , and 0.2 ml of virus dilution was added per dish. The dishes were tilted to spread the virus evenly, and adsorption was carried out for 30- to 60 minutes in a 37 degree Celsius incubator.
  • Overlay 10 ml per dish, consisted of a 1 : 1 ratio of 1 % (w/v) agarose in sterile water and 2X MEM Fll (supplemented with 8 ml of penicillin/streptomycin solution and 25 Unit/ml nystatin suspension). On 293 cells plaques were usually visible after 5-7 days and were visually counted.
  • the appropriate virus concentration was then diluted in PBS for oral installation into animals.
  • Recombinant vector AdHCMV13RG was evaluated for its ability to induce an immune response in striped skunks (Mephitis mephitis). Skunks were given a 2 ml dosage of virus DIOC by direct instillation into the mouth. At biweekly or monthly intervals, post immunization serum was tested for the production of rabies virus neutralizing antibodies (VNA). VNA determinations were carried out according to an ELISA developed by Barton and Campbell ((1988) J. Wildlife Dis. 24:246-258) or by a rabies fluorescence inhibition microtest (FIMT). Purified rabies glycoprotein was used as the antigen in an ELISA Assay.
  • FIMT equivalent units (E.U.) from a standard curve (Barton and Campbell, (1988) J. Wildlife Dis. 24:246-258).
  • One FIMT E.U. was defined as the inverse of the highest twofold serum dilution resulting in a reduction of fluorescent foci of ERA strain rabies virus grown under standard conditions by at least 50 %.
  • FIMT E.U. were divided by 46 to obtain international units ( U.)
  • FIMT EU International Units are calculated from FIMT equivalent Units by dividing by 46.
  • FIMT EU are based on an ELISA assay specific for rabies glycoprotein.

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  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Chemical & Material Sciences (AREA)
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  • Bioinformatics & Cheminformatics (AREA)
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Abstract

On décrit un adénovirus recombiné, conçu pour augmenter, chez un mammifère, une réponse immune à la rage, cet adénovirus présentant une région e1 interrompue fonctionnellement et comprenant une cassette d'expression dans laquelle l'ADN codant pour la glycoprotéine de la rage est lié de manière opérante à des commandes d'expression fonctionnelles chez un hôte eucaryote.
PCT/CA1996/000665 1995-10-05 1996-10-04 Adenovirus recombine de la rage WO1997012981A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
AU70818/96A AU7081896A (en) 1995-10-05 1996-10-01 Rabies recombinant adenovirus
EP96931711A EP0859846A1 (fr) 1995-10-05 1996-10-04 Adenovirus recombine de la rage

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US53941495A 1995-10-05 1995-10-05
US08/539,414 1995-10-05

Publications (1)

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WO1997012981A1 true WO1997012981A1 (fr) 1997-04-10

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PCT/CA1996/000665 WO1997012981A1 (fr) 1995-10-05 1996-10-04 Adenovirus recombine de la rage

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EP (1) EP0859846A1 (fr)
AU (1) AU7081896A (fr)
CA (1) CA2233983A1 (fr)
WO (1) WO1997012981A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000006751A3 (fr) * 1998-07-30 2000-05-04 Advec Inc Lignees cellulaires servant a produire des proteines represseurs pour la propagation de vecteurs
WO2003048198A3 (fr) * 2001-12-07 2004-04-22 Pasteur Institut Polypeptides induisant l'apoptose, polynucleotides les codant et leurs applications therapeutiques

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991002804A1 (fr) * 1989-08-22 1991-03-07 Ov Limited Sequence d'adenovirus recombinant exprimant l'antigene du rhabdovirus et son utilisation comme vaccin
WO1996039178A1 (fr) * 1995-06-05 1996-12-12 The Wistar Institute Of Anatomy And Biology Recombinant humain du type 5 de l'adenovirus a deficience de replication, utilise comme porteur de vaccins

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991002804A1 (fr) * 1989-08-22 1991-03-07 Ov Limited Sequence d'adenovirus recombinant exprimant l'antigene du rhabdovirus et son utilisation comme vaccin
WO1996039178A1 (fr) * 1995-06-05 1996-12-12 The Wistar Institute Of Anatomy And Biology Recombinant humain du type 5 de l'adenovirus a deficience de replication, utilise comme porteur de vaccins

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
GRAHAM F L ET AL: "METHODS FOR CONSTRUCTION OF ADENOVIRUS VECTORS", MOLECULAR BIOTECHNOLOGY, vol. 3, no. 3, 1995, pages 207 - 220, XP000603535 *
PREVEC L. ET AL.: "A recombinant human adenovirus vaccine against rabies", THE JOURNAL OF INFECTIOUS DISEASES, vol. 161, no. 1, January 1990 (1990-01-01), pages 27 - 30, XP000644392 *
XIANG Z. ET AL.: "A replication-defective human adenovirus recombinant serves as a highly efficacious vaccine carrier", VIROLOGY, vol. 219, no. 1, 1 May 1996 (1996-05-01), pages 220 - 227, XP002025004 *
XU Z. ET AL.: "Investigation of promoter function in human and animal cells infected with human recombinant adenoviruses expressing rotavirus antigen VP7sc", JOURNAL OF GENERAL VIROLOGY, vol. 76, no. 8, August 1995 (1995-08-01), pages 1971 - 1980, XP002025003 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000006751A3 (fr) * 1998-07-30 2000-05-04 Advec Inc Lignees cellulaires servant a produire des proteines represseurs pour la propagation de vecteurs
WO2003048198A3 (fr) * 2001-12-07 2004-04-22 Pasteur Institut Polypeptides induisant l'apoptose, polynucleotides les codant et leurs applications therapeutiques

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Publication number Publication date
AU7081896A (en) 1997-04-28
CA2233983A1 (fr) 1997-04-10
EP0859846A1 (fr) 1998-08-26

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