Method for the recovery of non-segmented, nagative-stranded RNA viruses from cDNA

A method for producing an infectious, non-segmented, negative-Stranded RNA virus of the Order Mononegavirales comprising the steps of:

• (a) introducing a viral cDNA expression vector comprising a polynucleotide encoding a genome or antigenome of a non-segmented negative-stranded RNA virus operably linked with an expression control sequence to direct synthesis of viral RNA transcripts from said cDNA expression vector into a host cell, wherein said host cell expresses an N protein, a P protein, and an L protein, and transiently expresses an RNA polymerase from a transiently-transfected expression vector;
• and
• (b) recovering assembled infectious, non-segmented, negative-stranded RNA virus from said host cells.

A method for producing an infectious, non-segmented, negative-stranded RNA virus of the Order Mononegavirales comprising the steps of:

• introducing into a suitable host cell a viral cDNA expression vector comprising a polynucleotide encoding a genome or antigenome of a non-segmented negative-stranded RNA virus operably linked with an expression control sequence to direct synthesis of viral RNA transcripts from said viral cDNA expression vector, and one or more transient expression vectors that encode(s) and direct(s) transient expression of an RNA polymerase, an N protein, a P protein, and an L protein;
• and
• recovering assembled infectious, non-segmented, negative-stranded RNA virus from said host cells.

An infectious, non-segmented, negative-stranded RNA virus of the Order Mononegavirales produced by the process of:

• introducing into a suitable host cell a viral cDNA expression vector comprising a polynucleotide encoding a genome or antigenome of a non-segmented negative-stranded RNA virus operably linked with an expression control sequence to direct synthesis of viral RNA transcripts from said cDNA expression vector, one or more support vector(s) that encode(s) and direct(s) expression of an N protein, a P protein, and an L protein, and a transient expression vector that encodes and directs transient expression of an RNA polymerase;
• and
• recovering assembled infectious, non-segmented, negative-stranded RNA virus from said host cells.

A method for stimulating the immune system of a mammalian subject to induce an immune response in the subject against a non-segmented negative-stranded RNA virus which comprises administering to the subject an immunogenically effective amount of an isolated, infectious, non-segmented, negative-stranded RNA virus produced according to the process of claim 39 in a pharmaceutically acceptable carrier.

A method for producing a growth- or replication-defective non-segmented, negative-stranded RNA virus of the Order Mononegavirales comprising the steps of:

• (a) introducing a viral cDNA expression vector comprising a polynucleotide encoding a genome or antigenome of a growth- or replication-defective non-segmented negative-stranded RNA virus operably linked with an expression control sequence to direct synthesis of viral RNA transcripts from said cDNA expression vector into a host cell, wherein said host cell expresses an N protein, a P protein, and an L protein, and transiently expresses an RNA polymerase from a transiently-transfected expression vector;
• and
• (b) recovering assembled growth- or replication-defective, non-segmented, negative-stranded RNA virus from said host cells.

A method for producing a growth- or replication-defective, non-segmented, negative-stranded RNA virus of the Order Mononegavirales comprising the steps of:

• introducing into a suitable host cell a viral cDNA expression vector comprising a polynucleotide encoding a genome or antigenome of a growth- or replication-defective non-segmented negative-stranded RNA virus operably linked with an expression control sequence to direct synthesis of viral RNA transcripts from said viral cDNA expression vector, and one or more transient expression vectors that encode(s) and direct(s) transient expression of an RNA polymerase, an N protein, a P protein, and an L protein;
• and
• recovering assembled growth- or replication-defective, non-segmented, negative-stranded RNA virus from said host cells.

A growth- or replication-defective, non-segmented, negative-stranded RNA virus of the Order Mononegavirales produced by the process of:

• introducing into a suitable host cell a viral cDNA expression vector comprising a polynucleotide encoding a genome or antigenome of a growth- or replication-defective non-segmented negative-stranded RNA virus operably linked with an expression control sequence to direct synthesis of viral RNA transcripts from said cDNA expression vector, one or more support vector(s) that encode(s) and direct(s) expression of an N protein, a P protein, and an L protein, and a transient expression vector that encodes and directs transient expression of an RNA polymerase;
• and
• recovering assembled growth- or replication-defective, non-segmented, negative-stranded RNA virus from said host cells.

An immunogenic composition comprising an immunogenically effective amount of an isolated, growth- or replication-defective, non-segmented, negative-stranded RNA virus produced according to the process of claim 98 in a pharmaceutically acceptable carrier.

A method for stimulating the immune system of a mammalian subject to induce an immune response in the subject against a non-segmented negative-stranded RNA virus which comprises administering to the subject an immunogenically effective amount of an isolated, growth- or replication-defective, non-segmented, negative-stranded RNA virus produced according to the process of claim 98 in a pharmaceutically acceptable carrier.