Abstract: The present invention relates to methods for intradermally delivering one or more biologically active agents such as vaccines and therapeutic agents into the dermis layer of the skin of a subject to obtain systemic delivery or an immune response using a microneedle drug delivery device containing the agent to be delivered. The methods employ a microneedle device with a row of hollow microneedles. The microneedles penetrate the skin of the subject and assume an anchored state in which the microneedles are anchored in the skin and project laterally from the device. A pivotal motion is then performed with the device so that the skin in which the microneedles are engaged is lifted above the initial plane of the surface of the skin while the biologically active agent is delivered. The methods of the invention elicit increased humoral and/or cellular response as compared to conventional vaccine delivery routes, facilitating dose sparing.

Abstract: This invention relates to a recombinant vector including a recombinant porcine adenovirus, stably incorporating and capable of expression of at least one heterologous nucleotide sequence. The nucleotide sequence is preferably one which encodes an antigenic determinant of Hog Cholera Virus or Pseudorabies virus. The further invention relates to a method of production of recombinant vectors, to methods of preparation of vaccines based on the vectors, to administration strategies and to methods of protecting pigs from disease.

Abstract: Agents and methods for enhancing recombinant virus transduction in the bladder epithelium are described. A first method involves contacting the luminal surface of the bladder with a composition comprising a transduction enhancing agent and an oncolytic virus. Alternatively, the luminal surface of the bladder can be contacted first with a pretreatment composition comprising a transduction enhancing agent and, subsequently, with a composition comprising an oncolytic virus. Bladder treatment compositions comprising a transduction enhancing agent and an oncolytic virus are also described.

Abstract: A modified virus including one or more non-native polypeptides, and the use of such viruses in therapy, particularly in the treatment of tumours or other cancerous cells are disclosed. The polypeptides includes one or more framework moieties each containing one or more binding moieties, and is capable of being expressed in the cytoplasm and nucleus of a mammalian host cell in a conformation which is maintained in the absence of a ligand for the binding moieties. The conformation allows the binding moieties subsequently to bind with the ligand, and the polypeptide is capable of transport through the nuclear membrane, wherein the modified virus has an altered tropism conferred by the binding moieties.

Abstract: The invention relates to viral formulations and related pharmaceutical products for use in gene therapy and/or vaccine applications. Especially preferred viral formulations disclosed herein are liquid adenovirus formulations, which show improved stability when stored in about the 2-8° C. range while also being compatible with parenteral administration. These formulations comprise a buffer, a sugar, a salt, a divalent cation, a non-ionic detergent, as well as a free radical scavenger and/or a chelating agent to inhibit free radical oxidation.

Abstract: The present invention includes a method to determine the immune status of an animal that includes the steps of (a) contacting a biological specimen of the animal with a recombinant infectious agent antigen that is specific for detecting an antibody selective for that infectious agent, under conditions suitable for formation of a complex between the recombinant antigen and the antibody and (b) detecting the presence or absence of the complex, wherein presence or absence of a complex is indicative of the immune status of the animal. Preferably such a method indicates whether the animal should be vaccinated. The present invention also includes an assay comprising (a) a recombinant infectious agent antigen that is specific for detecting an antibody selective for that infectious agent; and (b) a means to detect an antibody that selectively binds to the recombinant antigen.

Abstract: The present invention addresses the need to improve the yields of viral vectors when grown in cell culture systems. In particular, it has been demonstrated that for adenovirus, the use of low-medium perfusion rates in an attached cell culture system provides for improved yields. In other embodiments, the inventors have shown that there is improved Ad-p53 production witrh cells grown in serum-free conditions, and in particular in serum-free suspension culture. Also important to the increase of yields is the use of detergent lysis. Combination of these aspects of the invention permits purification of virus by a single chromatography step that results in purified virus of the same quality as preparations from double CsCl banding using an ultracentrifuge.

Abstract: The present invention relates to the field of biological engineering technology, specifically, to a SARS virus vaccine with adenovirus carrier, preparation method thereof and use of SARS virus S gene for preparation of severe acute respiratory syndrome (SARS) virus vaccine.

Abstract: An immunogenic regimen is provided. The regimen involves sequential administration of a recombinant adenoviral vector and a recombinant adeno-associated viral vector, each of which delivers a heterologous expression cassette encoding the same immunogenic product, or a cross-reactive immunogenic product. Also provided are products containing the vectors for use in the regimen of the invention.

Abstract: It is intended to provide a safe and efficient method of producing a virus which is free from animal-origin components in the whole process from culturing adhesive cells to producing the virus on an industrial scale by the cell culture. Namely, a method of producing a virus comprising: adhering adhesive cells to a support which has a polypeptide (P) having at least one cell-adhesive minimum amino acid sequence (X) per molecule, and is free from animal-origin components; culturing the adhesive cells in a medium free from animal-origin components; subculturing the cultured adhesive cells using a cell dispersing agent free from animal-origin components; and then inoculating and proliferating a virus in the cells obtained by culturing the adhesive cells.

Abstract: The present invention is intended to provide a pharmaceutical composition for delivering a chemotherapeutic, preferably an anticancer drug, into cells or into a living organism, using a viral envelope vector, and provides a pharmaceutical composition comprising a chemotherapeutic encapsulated in, or used in combination with, a viral envelope vector having an adjuvanticity as an active ingredient. Thereby it is possible to introduce an anticancer drug encapsulated in a viral envelope vector directly into a tumor, with coadministration of another anticancer drug so as to induce tumor cell-specific antitumor immunity also thanks to the adjuvant action of HVJ-E, and hence to regress the tumor. The present invention also provides a pharmaceutical composition comprising a viral envelope vector and a chemotherapeutic as active ingredients.

Abstract: A vaccine composition for vaccinating dogs comprising any one or more of (a) an agent capable of raising an immune response against Streptococcus equi sub species zooepidemicus in a dog, (b) an agent capable of raising an immune response against Mycoplasma cynos in a dog, and (c) an agent capable of raising an immune response against a Chlamydophila in a dog.

Abstract: Disclosed is a method of modulating neutralizing antibodies formation against a heterologous protein. The method may be used to induce tolerance of the immune system towards the protein, such tolerance being useful to allow long-term gene therapy or transgene expression. The method may also be used to provide an animal with a reproducible functional inactivation phenotype of an endogenous protein of the animal.

Abstract: The present invention addresses the need to improve the yield of adenovirus when grown in cell culture systems. In particular, it has been demonstrated that for adenovirus, the use of infection temperatures lower than 37° C. in a cell culture system results in improved yields of adenovirus. In addition, it has been demonstrated that when host cells are grow in a bioreactor, initiating adenovirus infection by diluting the host cells with fresh media and adenovirus results in improved yield of adenovirus. Methods of adenoviral production and purification using infection temperatures less than 37° C. are disclosed. Methods of adenoviral production and purification wherein the host cells are grown in a bioreactor and adenovirus infection is initiated by diluting the host cells with fresh media and adenovirus are also disclosed.

Abstract: The present invention provides nucleotide sequences from SARS (Serve Acute Respiratory Syndrome) coronavirus genomes, as well as the applications of the partial fragments thereof in preparing DNA vaccine or expressing corresponding proteins. Furthermore, the present invention also provides the uses of said proteins in preventing and treating diseases, and preparing antibodies.

Abstract: Disclosed herein are novel composition and methods for altering the proliferation of a cell. Included are wild-type and mutant hKIS polypeptides along with cyclin kinase inhibitors containing mutations that prevent their inhibition with serine/threonine kinases.

Abstract: This invention provides novel replication-defective adenoviral vectors comprising an adenoviral genome in which the protein IX gene, preferably under the control of its own promoter, is in an inverted orientation relative to the direction of transcription of the native protein IX gene at a location where the protein IX gene normally resides, for production of replication-competent adenovirus (RCA) free, or substantially RCA-free, adenovirus preparations. Said vector preferably encodes a gene of interest. The invention relates to viral particles, host cells and compositions comprising said adenoviral vector. This invention further relates a method for propagating adenovirus preparations, free, or substantially free, of replication-competent adenovirus (RCA) particles, from host cells comprising vectors of this invention, for use to treat a subject suffering from a disease or disorder or to prevent a subject from getting a disease or disorder, such as cancer.

Abstract: The present invention provides an adenovirus serotype 30 (Ad30) fiber amino acid sequence. The present invention also provides polynucleotides and expression vectors encoding an Ad30 fiber and viral particles and cells containing such expression vectors. The present invention further provides methods of treating genetic diseases or cancers in a mammal using the polynucleotides, polypeptides, expression vectors, viral particles and cells of the present invention.

Abstract: The present disclosure provides compositions and methods for eliciting an immune response against avian or pandemic influenza. The compositions include adenovirus vectors comprising avian influenza antigens, recombinant adenovirus and immunogenic compositions comprising such recombinant vectors and adenovirus. Methods for eliciting an immune response against avian or pandemic influenza involving administering such adenovirus vectors or recombinant adenovirus are also provided.

Abstract: The invention relates to novel vaccine regimens in which specific prime/boost regimens are applied using low-neutralized recombinant adenoviral vectors harboring nucleic acids encoding antigens from Plasmodium falciparum and purified recombinant protein vaccines such as RTS,S, in the context of appropriate adjuvants.

Abstract: Provided are adenovirus vectors and the production of such vectors. In particular, adenoviruses with modified or heterologous fiber proteins for targeting to dendritic cells are provided.

Abstract: The invention relates to viral formulations and related pharmaceutical products for use in gene therapy and/or vaccine applications. Especially preferred viral formulations disclosed herein are liquid adenovirus formulations, which show improved stability when stored in about the 2-8° C. range while also being compatible with parenteral administration. These formulations comprise a buffer, a sugar, a salt, a divalent cation, a non-ionic detergent, as well as a free radical scavenger and/or a chelating agent to inhibit free radical oxidation.

Abstract: A simple yet effective method of increasing production of a thermo-stable virus, such as adenovirus and picornavirus, is presented. The method entails a temperature shift strategy whereby the culture of host cells are shifted to a sub-optimal temperature for a period of time prior to virus infection or cells are grown at a sub-optimal level for the entire cell expansion process including one or more than one passages of cell growth from cryopreserved cells, followed by a shift back to a more optimal temperature at or near the time of virus infection of the respective host cells. Adaptation of such a temperature shift strategy present a simple yet effective method to substantially increase recoverable virus within a respective host cell/virus production scheme without the need to further manipulate other culture and/or media conditions within an established host cell/virus production scheme.

Abstract: The complete nucleotide sequence of the genome of porcine adenovirus type 3 (PAV-3) is provided. Methods for construction of infectious PAV genomes by homologous recombination in procaryotic cells are provided. Recombinant PAV viruses are obtained by transfection of mammalian cells with recombinant PAV genomes. The PAV-3 genome can be used as a vector for the expression of heterologous nucleotide sequences, for example, for the preparation and administration of subunit vaccines to swine or other mammals. In addition, PAV-3 vectors can be used for gene therapy and expression of heterologous polypeptides. PAV-3 genome sequences can also be used for diagnostic purposes, to detect the presence of PAV-3 DNA in a subject or biological sample.

Abstract: A process for purifying virus particles, especially recombinant adenovirus vector particles, is presented. The process relies on various combinations of cell lysis, detergent-based precipitation of host cell contaminants away from the virus, depth filtration or centrifugation, ultrafiltration, nuclease digestion and chromatography to robustly and economically produce highly purified product. This process results in contaminating DNA levels which are consistently below detectable levels.

Abstract: This invention relates to a recombinant vector including a recombinant porcine adenovirus, stably incorporating and capable of expression of at least one heterologous nucleotide sequence. The nucleotide sequence is preferably one which encodes an antigenic determinant of Hog Cholera Virus or Pseudorabies virus. The further invention relates to a method of production of recombinant vectors, to methods of preparation of vaccines based on the vectors, to administration strategies and to methods of protecting pigs from disease.

Abstract: The invention concerns nucleic acids derived from human V9 erythrovirus, fragments thereof, encoded polypeptides, and applications as diagnostic reagents and as immunogenic agents.

Abstract: The present invention relates to foreign peptide sequences fused to recombinant plant viral structural proteins and a method of their production. Fusion proteins are economically synthesized in plants at high levels by biologically contained tobamoviruses. The fusion proteins of the invention have are useful as antigens for inducing the production of antibodies having desired binding properties, e.g., protective antibodies, or for use as vaccine antigens for the induction of protective immunity against the parvovirus. Feline parvovirus epitopes were fused to the N-terminus of the TMV coat protein, expressed in Nicotiana plants, extracted, purified, characterized and administered to animals, resulting in protective immunity.

Abstract: Agents and methods for enhancing recombinant virus transduction in the bladder epithelium are described. A first method involves contacting the luminal surface of the bladder with a composition comprising a transduction enhancing agent and an oncolytic virus. Alternatively, the luminal surface of the bladder can be contacted first with a pretreatment composition comprising a transduction enhancing agent and, subsequently, with a composition comprising an oncolytic virus. Bladder treatment compositions comprising a transduction enhancing agent and an oncolytic virus are also described.

Abstract: This invention relates to a parvovirus vector having a parvovirus DNA excisable from the vector DNA in a parvovirus-permissive cell, the parvovirus DNA having a left terminus which comprises a parvovirus minimal origin of replication, and a system comprising the parvovirus vector. Furthermore, this invention concerns a method of producing the parvovirus vector, parvoviral particles as well as their use.

Abstract: The invention concerns a method for purifying a crude viral preparation containing viral, in particular adenoviral, particles of interest. The invention is characterised in that it comprises a step of adsorption on a fluidised bed. The invention also concerns a protocol for producing viral particles for use in gene therapy comprising such a purifying process.

Abstract: The present invention relates to the identification of bovine adenovirus sequence(s) essential for encapsidation and E1 transcriptional control regions. The present invention provides adenovirus expression systems, host cells and compositions comprising adenovirus vectors which comprise one or more BAV sequence(s) essential for encapsidation, as well as helper virus which express BAV sequences essential for encapsidation. The present invention also provides helper vectors comprising a BAV sequence essential for encapsidation which is used in a helper virus for propagating recombinant adenovirus. The present invention also provides adenovirus expression systems, host cells and compositions comprising adenovirus vectors which comprise modifications in BAV E1 transcriptional control regions. The present invention also provides methods for making adenovirus vectors comprising BAV sequence(s) essential for encapsidation as well as modifications in BAV E1 transcriptional control regions.

Abstract: Methods of making and using recombinant AAV virions with decreased immunoreactivity are described. The recombinant AAV virions include mutated capsid proteins or are derived from non-primate mammalian AAV serotypes and isolates that display decreased immunoreactivity relative to AAV-2.

Abstract: The invention relates to a structural protein of adeno-associated virus (AAV) that contains at least one modification that reduces the antigenicity of the virus. The invention further relates to the production of the structural protein and to its use.

Abstract: The invention provides an isolated and purified DNA molecule comprising at least one DNA segment, a biologically active subunit or variant thereof, of a circular intermediate of adeno-associated virus, which DNA segment confers increased episomal stability, persistence or abundance of the isolated DNA molecule in a host cell. The invention also provides a composition comprising at least two adeno-associated virus vectors.

Abstract: The present invention is based upon the finding that the protein p300 has acetylation activity which is directed to the retinoblastoma tumour suppressor protein pRb by the presence in the cell of the adenovirus E1A protein. This represents a target for modulators of the cell cycle, to which end the invention provides an assay for a modulator of acetylation of pRb by p300, which comprises: a) bringing into contact a p300 protein a pRb protein and a putative modulator compound under conditions where the p300 protein, in the absence of said modulator is capable of acetylating the pRb protein; b) providing conditions for acetylation of said pRb protein; and c) measuring the degree of inhibition of acetylation caused by said modulator compound.

Abstract: The present invention provides packaging cell lines for the efficient production of an Adeno-associated virus (AAV) vector which does not require “helper” virus function for the replication and encapsidation of the AAV vector particles. Packaging cells, methods for their production and methods for producing recombinant AAV vector particles useful for human gene therapy are provided.

Abstract: The present invention relates to methods for culturing circovirus and in particular, porcine circovirus. The present invention provides compositions and methods for culturing porcine circovirus in mammalian cells expressing mammalian adenovirus E1 function.

Abstract: The present invention provides methods and compositions useful in localized transfer of genetic material or proteins. Moreover, the present invention provides methods and compositions for improving and/or controlling wound healing by applying a wound care device comprising HoxD3 and/or HoxA3 and/or HoxB3. In addition, the present invention provides methods and compositions for improved wound healing in subjects having impaired healing capabilities, such as diabetic subjects.

Abstract: The present invention relates to an AAV DNA having helper virus sequences which are necessary for developing AAV viral particles, a system containing such a DNA and the use of both.

Abstract: This invention provides methods and compositions to preserve bioactive materials in a dried foam matrix. Methods provide non-boiling foam generation and penetration of preservative agents at temperatures near the phase transition temperature of the membranes.

Abstract: The present invention relates to methods for efficient and reliable construction of adenovirus vectors which contain and express foreign DNA and are useful for gene transfer into mammalian cells, for vaccines and for gene therapy. The invention provides for the growth and purification of adenovirus vectors (helper dependent vectors or HDVs) from which all or most of the viral genes have been removed. The vector system described herein is a new method designed to eliminate helper viruses from the final HDV preparation by cleavage of the helper virus DNA with an endonuclease, alone or in combination with other methods known to limit the level of helper virus contamination of helper dependent vector preparations. The disclosed methods and compositions also provide for regulated control of gene expression.

Abstract: Methods and compositions for the production of recombinant proteins in a human cell line. The methods and compositions are particularly useful for generating stable expression of human recombinant proteins of interest that are modified post-translationally, for example, by glycosylation. Such proteins may have advantageous properties in comparison with their counterparts produced in non-human systems such as Chinese hamster ovary cells.

Abstract: The present invention addresses the need to improve the yields of viral vectors when grown in cell culture systems. In particular, it has been demonstrated that for adenovirus, the use of low-medium perfusion rates in an attached cell culture system provides for improved yields. In other embodiments, the inventors have shown that there is improved Ad-p53 production cells grown in serum-free conditions, and in particular in serum-free suspension culture. Also important to the increase of yields is the use of detergent lysis. Combination of these aspects of the invention permits purification of virus by a single chromatography step that results in purified virus of the same quality as preparations from double CsCl banding using an ultracentrifuge.

Abstract: The present invention provides methods and compositions useful in localized transfer of genetic material or proteins. Moreover, the present invention provides methods and compositions for improving and/or controlling wound healing by applying a wound care device comprising HoxD3 and/or HoxA3. In addition, the present invention provides methods and compositions for improved wound healing in subjects having impaired healing capabilities, such as diabetic subjects.

Abstract: An in vivo replicative and recombined porcine adenovirus characterized in that it comprises a heterologous nucleotide sequence inserted into the porcine adenovirus in conditions enabling the latter to be replicated in vivo and to express the inserted heterologous nucleotide sequence, and in that the adenovirus genome comes from a 3 or 5 serotype (PAV-3 or PAV-5) adenovirus. Insertion occurs in a non-essential zone of the E3 region, preferably with deletion of said zone. The invention also relates to a recombined porcine vaccine comprising one such porcine adenovirus. The invention further relates to a serotype 3 or 5 porcine adenovirus vector that is replicative in vivo and is deleted in a non-essential region of the genome thereof. The invention also relates to a DNA fragment comprising all or part of the referenced SEQ ID NO.5 nucleotide sequence.

Abstract: A viral DNA construct, and virus encoded thereby, is provided having one or more tumour specific transcription factor binding sites in place of one or more wild type transcription factor binding sites operatively positioned in the promoter region which controls expression of early genes responsible for viral nucleic acid replication. Preferred constructs place the tumour specific transcription factor binding sites in operative relation to DNA polymerase, DNA terminal protein and/or DNA binding protein. Compositions and constructs contained therein are provided, particularly for use in therapy. Methods of treating patients for neoplasms are also provided.

Abstract: The present invention provides delivery vectors for transferring a nucleic acid sequence to a cell in vitro, ex vivo or in vivo. The delivery vector comprises a segment encoding a secretory signal peptide. In embodiments of the invention, the delivery vector is an adeno-associated virus (AAV) vector. In other embodiments, the secretory signal peptide is a fibronectin secretory signal peptide (including variations and modifications, thereof). The delivery vectors of the invention may further comprise a heterologous nucleic acid sequence encoding a polypeptide of interest for transfer to a target cell, where the polypeptide of interest is operably associated with the secretory signal. Also disclosed are methods of transferring a nucleic acid of interest to a cell using the delivery vectors of the invention.

Abstract: The present invention provides a vaccine accelerator factor (VAF) which is an in ovo nucleotide immuno-stimulant. The VAF contains one or more DNA constructs, each having a DNA molecule and a vector. Each of the DNA molecule contains one or more genes or gene fragments, each encoding an antigenic peptide of an avian virus. The VAF is preferably administered to the amniotic fluid of an egg after being fertilized for about 17–19 days. The VAF can be co-administered with a viral vaccine containing one or more attenuated or inactive avian viruses. Alternatively, the VAF can be administered prior to the administration of the viral vaccine, which is administered at hatch or post-hatch. The VAF stimulates and accelerate a protective immune response of a viral vaccine.

Abstract: The present invention provides a modified adenovirus comprising genomic adenoviral DNA which has been modified so that (i) the only gene product of the early region (E4) that is expressed is open reading frame 6 (ORF-6), (ii) neither the gene product of the E1A region nor the gene product of the E1B region is expressed, and (iii) no other early or late gene products are expressed. The present invention also provides methods of inhibiting repair of breaks in double-stranded DNA in a cell, preventing concatamerization of linear wild-type adenoviral DNA, inhibiting V(D)J recombination of nucleic acid sequences encoding immunoglobulins, preventing apoptosis, and preventing and treating cancer.