Abstract: The invention provides recombinant viruses comprising heterologous envelope proteins capable of mediating entry of the recombinant viruses into host cells. In one embodiment, a recombinant virus of the invention is a member of the Paramyx-ovirzdae family (e.g., a respiratory syncytial virus), and the heterologous envelope protein includes the ectodomain of a baculovirus envelope protein (e.g., the GP64 protein). In some cases, the heterologous envelope protein is provided in addition to homologous envelope proteins) which may or may not be functional. The heterologous protein can provide the recombinant virus with enhanced stability (e.g., at 4° C., 22° C., or 37° C.), and allows production of high-titer virus stocks. The heterologous protein can also impart temperature sensitivity to the replication of the recombinant virus. In addition, the recombinant virus can be designed to be infectious but incapable of spreading between host cells by providing the heterologous protein by complementation in trans.

Abstract: The present invention provides recombinant respiratory syncytial viruses (RSV) in which all of the surface glycoprotein genes encoding the attachment protein G, the fusion protein F, and the Small Hydrophobic protein SH are deleted. The genes are replaced by a chimeric gene encoding a heterologous entry protein derived from the Vesicular Stomatitis Virus G protein or GP64 of baculovirus. Alternatively, the replacement proteins are provided in trans. Marker genes such as those encoding ?-glucuronidase (GUS) and green fluorescent protein (EGFP) are also added to the upstream and downstream side of the hybrid gene for easy detection. These infectious recombinant respiratory syncytial viruses offer alternatives and improvements as vaccine candidates.

Abstract: The present invention provides a method of increasing expression of a promoter distal gene in a virus of the order Mononegavirales, and a recombinant virus constructed by such method. Also provided is a method of attenuating a virus of the order Mononegavirales, and of constructing an attenuated virus useful for a vaccine.

Abstract: The present invention relates to recombinant DNA molecules which encode bovine respiratory syncytial (BRS) virus proteins, to BRS viral proteins, and peptides and to recombinant BRS virus vaccines produced therefrom. It is based, in part, on the cloning of substantially full length cDNAs which encode the entire BRS virus G, F, and N proteins. According to particular embodiments of the invention, DNA encoding a BRS virus protein or peptide may be used to diagnose BRS virus infection, or, alternatively, may be inserted into an expression vector, including, but not limited to, vaccinia virus as well as bacterial, yeast, insect, or other vertebrate vectors. These expression vectors may be utilized to produce the BRS virus protein or peptide in quantity; the resulting substantially pure viral peptide or protein may be incorporated into subunit vaccine formulations or may be used to generate monoclonal or polyclonal antibodies which may be utilized in diagnosis of BRS virus infection or passive immunization.

Abstract: The present invention provides a method of increasing expression of a promoter distal gene in a virus of the order Mononegavirales, and a recombinant virus constructed by such method. Also provided is a method of attenuating a virus of the order Mononegavirales, and of constructing an attenuated virus useful for a vaccine.

Abstract: The present invention relates to recombinant DNA molecules which encode bovine respiratory syncytial (BRS) virus proteins, to BRS virus proteins, and peptides and to recombinant BRS virus vaccines produced therefrom. It is based, in part, on the cloning of substantially full length cDNAs which encode the entire BRS virus G, F, and N proteins. According to particular embodiments of the invention, DNA encoding a BRS virus protein or peptide may be used to diagnose BRS virus infection, or, alternatively, may be inserted into an expression vector, including, but not limited to, vaccinia virus as well as bacterial, yeast, insect, or other vertebrate vectors. These expression vectors may be utilized to produce the BRS virus protein or peptide in quantity; the resulting substantially pure viral peptide or protein may be incorporated into subunit vaccine formulations or may be used to generate monoclonal or polyclonal antibodies which may be utilized in diagnosis of BRS virus infection or passive immunization.

Abstract: The present invention provides a method of increasing expression of a promoter distal gene in a virus of the order Mononegavirales, and a recombinant virus constructed by such method. Also provided is a method of attenuating a virus of the order Mononegavirales, and of constructing an attenuated virus useful for a vaccine.

Abstract: The present invention provides a method of increasing expression of a promoter distal gene in a virus of the order Mononegavirales, and a recombinant virus constructed by such method. Also provided is a method of attenuating a virus of the order Mononegavirales, and of constructing an attenuated virus useful for a vaccine.

Abstract: The present invention demonstrates that the M2-1 protein of respiratory syncytial virus has a conserved Cys3—His1 motif known to bind zinc ions in other proteins and that mutations of the predicted zinc coordinating residues in the Cys3—His1 motif affect the transcriptional antitermination activity of M2-1, its ability to interact with nucleocapsid protein, and the phosphorylation state of M2-1. This invention clearly demonstrates the requirement for conservation of the Cys3—His1 motif in order to maintain the functional integrity of the M2-1 protein. Therefore, the present invention provides for methods of designing and screening compounds for antiviral activity towards respiratory syncytial virus based upon the loss of function of the M2-1 protein.

Abstract: The present invention demonstrates that the M2-1 protein of respiratory syncytial virus has a conserved Cys3-His1 motif known to bind zinc ions in other proteins and that mutations of the predicted zinc coordinating residues in the Cys3-His1 motif affect the transcriptional antitermination activity of M2-1, its ability to interact with nucleocapsid protein, and the phosphorylation state of M2-1. This invention clearly demonstrates the requirement for conservation of the Cys3-His1 motif in order to maintain the functional integrity of the M2-1 protein. Therefore, the present invention provides for methods of designing and screening compounds for antiviral activity towards respiratory syncytial virus based upon the loss of function of the M2-1 protein.

Abstract: Provided is a method of attenuating a virus of the order Mononegavirales, comprising the step of: rearranging said virus' gene order by moving a gene essential for RNA replication away from its wild-type 3' promoter proximal position site, wherein said gene is an essential limiting factor for genome replication and is placed in the next to last position in the gene order. Also provided is a method of making an attenuated virus useful for a vaccine, comprising the steps of: rearranging said virus' gene order by moving a gene essential for RNA replication away from its wild-type 3' promoter proximal position site, wherein a gene which is a n essential limiting factor for genome replication is placed in the next to last position in the gene order; and placing a gene coding for an immune response inducing antigen in the position closest to the 3' end of the gene order. Also provided is a method of attenuating a virus of the order Mononegavirales, comprising the step of: rearranging said virus' gene order.

Abstract: The present invention relates to recombinant DNA molecules which encode bovine respiratory syncytial (BRS) virus proteins, to BRS virus proteins, and peptides and to recombinant BRS virus vaccines produced therefrom. It is based, in part, on the cloning of substantially full length cDNAs which encode the entire BRS virus C, F, and N proteins. According to particular embodiments of the invention, DNA encoding a BRS virus protein or peptide may be used to diagnose BRS virus infection, or, alternatively, may be inserted into an expression vector, including, but not limited to, vaccinia virus as well as bacterial, yeast, insect, or other vertebrate vectors. These expression vectors may be utilized to produce the BRS virus protein or peptide in quantity; the resulting substantially pure viral peptide or protein may be incorporated into subunit vaccine formulations or may be used to generate monoclonal or polyclonal antibodies which may be utilized in diagnosis of BRS virus infection or passive immunization.

Abstract: Recombinant methods for recovering wildtype or engineered negative stranded, non-segmented RNA virus genomes containing non-coding 3' and 5' regions (e.g. leader or trailer regions) surrounding one, several or all of the genes of the virus or one or more heterologous gene(s) in the form of ribonucleocapsids containing N, P and L proteins, which are capable of replicating and assembling with the remaining structural proteins to bud and form virions, or which are only capable of infecting one cell, or are transcribing particles, are disclosed. Novel vaccines, gene therapy vectors and antiviral compounds based on these viral particles are also disclosed.

Abstract: Recombinant methods for recovering wildtype or engineered negative stranded, non-segmented RNA virus genomes containing non-coding 3' and 5' regions (e.g. leader or trailer regions) surrounding one, several or all of the genes of the virus or one or more heterologous gene(s) in the form of ribonucleocapsids containing N, P and L proteins, which are capable of replicating and assembling with the remaining structural proteins to bud and form virions, or which are only capable of infecting one cell, or are defective interfering particles, are disclosed. Novel vaccines, gene therapy vectors and antiviral compounds are also disclosed.

Abstract: This invention discloses compositions of DNA and proteins that are useful for preparing vaccines against human respiratory syncytial virus ?HRSV!. The DNA compositions include structural genes coding for native structural viral proteins and immunogenic fragments of these proteins. Host cells transformed with the above DNA compositions are also disclosed. Vaccines made from the native structural viral proteins or immunogenic fragments are also disclosed as well as methods for protecting humans by inoculation with these vaccines.

Abstract: This invention discloses compositions of DNA and proteins that are useful for preparing vaccines against human respiratory syncytial virus [HRSV]. The DNA compositions include structural genes coding for native structural viral proteins and immunogenic fragments of these proteins. Host cells transformed with the above DNA compositions are also disclosed. Vaccines made from the native structural viral proteins or immunogenic fragments are also disclosed as well as methods for protecting humans by inoculation with these vaccines.