NOVEL METHODS AND INTERFERON DEFICIENT SUBSTRATES FOR THE PROPAGATION OF VIRUSES
The present invention relates, to novel methods and substrates for the propagation of viruses. The invention relates to IFN-deficient substrates and methods for propagating viruses in these unconventional substrates. In particular, the invention relates to methods of propagating viruses in immature embryonated eggs, preferably six- to nine-day-old chicken eggs. The methods of the invention are particularly attractive for growing viruses suitable for use in vaccine and pharmaceutical formulations.
1 . An interferon deficient cell line containing a genetically engineered attenuated influenza virus, in which the genome of the attenuated influenza virus encodes a truncated NS1 protein composed of between 90 and 130 N-terminal amino acid residues of the NS1 protein of the same or a different influenza virus strain, so that the genetically engineered influenza virus has an impaired interferon antagonist phenotype.
2 . The interferon deficient cell line of claim 1 in which the genetically engineered attenuated influenza virus genome encodes a truncated NS1 protein composed of between 90 and 100 N-terminal amino acid residues of the NS1 protein of the same or a different influenza virus strain.
3 . The interferon deficient cell line of claim 1 in which the genetically engineered attenuated influenza virus genome encodes a truncated NS1 protein of between 100 and 110 N-terminal amino acid residues of the NS1 protein of the same or a different influenza virus strain.
4 . The interferon deficient cell line of claim 1 in which the genetically engineered attenuated influenza virus genome encodes a truncated NS1 protein of between 110 and 120 N-terminal amino acid residues of the NS1 protein of the same or a different influenza virus strain.
5 . The interferon deficient cell line of claim 1 in which the genetically engineered attenuated influenza virus genome encodes a truncated NS1 protein of between 120 and 130 N-terminal amino acid residues of the NS1 protein of the same or a different influenza virus strain.
6 . An interferon deficient cell line containing a genetically engineered attenuated influenza virus, in which the genome of the attenuated influenza virus encodes a truncated NS1 protein composed of amino acid residues 1 to 130, amino acid residues 1 to 124, amino acid residues 1 to 120, amino acid residues 1 to 110, amino acid residues 1 to 100, amino acid residues 1 to 99, amino acid residues 1 to 90, amino acid residues 1 to 89, amino acid residues 1 to 70, or amino acid residues 1 to 60 of the NS1 protein of the same or a different influenza virus strain, so that the genetically engineered attenuated influenza virus has an impaired interferon antagonist phenotype.
7 . The interferon deficient cell line of claim 6 in which the genetically engineered influenza virus genome encodes a truncated NS1 protein composed of amino acid residues 1 to 130 of the NS1 protein of the same or a different influenza virus strain.
8 . The interferon deficient cell line of claim 6 in which the genetically engineered influenza virus genome encodes a truncated NS1 protein composed of amino acid residues 1 to 124 of the NS1 protein of the same or a different influenza virus strain.
9 . The interferon deficient cell line of claim 6 in which the genetically engineered influenza virus genome encodes a truncated NS1 protein composed of amino acid residues 1 to 120 of the NS1 protein of the same or a different influenza virus strain.
10 . The interferon deficient cell line of claim 6 in which the genetically engineered influenza virus genome encodes a truncated NS1 protein composed of amino acid residues 1 to 110 of the NS1 protein of the same or a different influenza virus strain.
11 . The interferon deficient cell line of claim 6 in which the genetically engineered influenza virus genome encodes a truncated NS1 protein composed of amino acid residues 1 to 100 of the NS1 protein of the same or a different influenza virus strain.
12 . The interferon deficient cell line of claim 6 in which the genetically engineered influenza virus genome encodes a truncated NS1 protein composed of amino acid residues 1 to 99 of the NS1 protein of the same or a different influenza virus strain.
13 . The interferon deficient cell line of claim 6 in which the genetically engineered influenza virus genome encodes a truncated NS1 protein composed of amino acid residues 1 to 90 of the NS1 protein of the same or a different influenza virus strain.
14 . The interferon deficient cell line of claim 6 in which the genetically engineered influenza virus genome encodes a truncated NS1 protein composed of amino acid residues 1 to 89 of the NS1 protein of the same or a different influenza virus strain.
15 . The interferon deficient cell line of claim 6 in which the genetically engineered influenza virus genome encodes a truncated NS1 protein composed of amino acid residues 1 to 70 of the NS1 protein of the same or a different influenza virus strain.
16 . The interferon deficient cell line of claim 6 in which the genetically engineered influenza virus genome encodes a truncated NS1 protein composed of amino acid residues 1 to 60 of the NS1 protein of the same or a different influenza virus strain.
17 . The interferon deficient cell line of claim 6 in which the interferon deficient cell line is not a STAT1 (−) cell line.
18 . The interferon deficient cell line of claim 17 in which the interferon deficient cell line is not Vero cells.
19 . The interferon deficient cell line of claim 1 in which the interferon deficient cell line is not Vero cells.
20 . The interferon deficient cell line of claim 6 in which the interferon deficient cell line is Vero cells.
21 . An interferon deficient cell line containing an attenuated influenza virus, in which the genome of the attenuated influenza virus has an NS1 gene of the same or a different influenza virus strain, wherein the NS1 gene has a mutation such that the attenuated influenza virus has an impaired interferon antagonist phenotype, wherein said virus is not influenza C virus and the interferon deficient cell line is not Vero cells and is not STAT1 (−) cell lines.
22 . The interferon deficient cell line of claim 21 , wherein the attenuated influenza virus is genetically engineered.
23 . The interferon deficient cell line of claim 21 , wherein the mutation in the NS1 gene is a deletion at the C-terminus of NS1.
24 . The interferon deficient cell line of claim 21 , wherein the NS1 gene encodes a truncated NS1 protein composed of amino acid residues 1 to 80 of the NS1 protein of the same or a different influenza virus strain.
25 . The interferon deficient cell line of claim 6 , wherein the influenza virus is an influenza A virus.
26 . The interferon deficient cell line of claim 6 , wherein the influenza virus is an influenza B virus.
27 . The interferon deficient cell line of claim 6 , wherein the attenuated influenza virus is engineered to encode a foreign antigen.
28 . The interferon deficient cell line of claim 6 , wherein the attenuated influenza virus is engineered to encode a tumor antigen.
29 . The interferon deficient cell line of claim 6 , wherein the attenuated influenza virus is engineered to encode an epitope from an infectious agent other than an influenza virus.
30 . The interferon deficient cell line of claim 6 , wherein the attenuated influenza virus is engineered to encode an epitope from another virus.
31 . The interferon deficient cell line of claim 6 , wherein the attenuated influenza virus is engineered to encode an epitope from a bacteria.
32 . The interferon deficient cell line of claim 6 , wherein the attenuated influenza virus is engineered to encode an epitope from a parasite.