Generation of infectious influenza viruses from virus-like particles
View Patent ↗Compositions and methods to prepare influenza virus-like particles (VLPs) are provided.
1. A method to prepare influenza A or B virus comprising:
infecting a host cell with two different VLPs, wherein a first isolated VLP has at least one and up to eight influenza A virus segments and a second VLP has at least one and up to eight influenza A virus viral segments, wherein the first VLP and the second VLP comprise one or up to eight influenza A virus viral segments selected from:
an influenza virus PA segment;
an influenza virus PB1 segment;
an influenza virus PB2 segment;
an influenza virus NP segment;
an influenza virus M segment;
an influenza virus NS segment;
an influenza virus NA segment; or
an influenza virus HA segment or a modified HA segment having sequences for a non-influenza host cell binding protein; or
wherein the first VLP and the second VLP comprise one or up to eight influenza B virus viral segments selected from:
an influenza virus PA segment;
an influenza virus PB1 segment;
an influenza virus PB2 segment;
influenza virus NP segment;
an influenza virus M segment;
an influenza virus NS segment;
an influenza virus NA segment; or
an influenza virus HA segment or a modified HA segment having sequences for a non-influenza host cell binding protein;
and
wherein the first VLP and the second VLP each comprise PA, PB1 PB2, NP, and HA or a non-influenza host cell binding protein;
wherein the first VLP has at least one viral segment that is not present in the second VLP or wherein the first VLP has at least one segment that is modified in the second VLP so that a functional influenza virus protein is not expressed from that modified segment; and
isolating progeny virus.
2. The method of claim 1 , wherein the second VLP has at least one viral segment that is not present in the first VLP.
3. The method of claim 1 , wherein at least one of the viral segments of the first VLP is from the same virus isolate as the corresponding viral segment in the second VLP.
4. The method of claim 1 , wherein at least one of the viral segments of the first VLP is from a different virus isolate as the corresponding segment in the second VLP.
5. The method of claim 1 , wherein the first VLP has 7 viral segments of or the second VLP has 7 viral segments; or wherein the first VLP has 7 viral segments and the second VLP has 6 viral segments; or wherein the first VLP has at least 3 viral segments and the second VLP has 7 viral segments; or wherein the first VLP has at least 4 viral segments and the second VLP has at least 4 viral segments; or wherein the first VLP has less than 8 viral segments and the second VLP has less than 8 viral segments.
6. The method of claim 1 , wherein the viral segments for HA and NA are from the same virus isolate.