Virus like particle comprising modified envelope protein E3
A virus like particle comprising a viral structural protein which comprises modified envelope protein E3. The viral structural protein may be that derived from or alphavirus or flavivirus. Especially, the viral structural protein may be derived from Chikungunya virus or Venezuelan equine encephalitis virus.
1. An alphavirus virus-like particle, wherein said virus-like particle comprises an alphaviral structural protein that comprises an envelope protein E3, wherein said envelope protein E3 is modified to contain at least one foreign antigen inserted into furin cleavage site thereof.
2. The virus-like particle according to claim 1 , wherein the alphaviral structural protein is a Chikungunya virus (CHIKV) structural protein or a Venezuelan equine encephalitis virus (VEEV) structural protein.
3. The virus-like particle according to claim 2 , wherein the alphaviral structural protein is a viral structural protein of Chikungunya virus strain 37997 or strain OPY-1, or is a viral structural protein of Venezuelan equine encephalitis virus strain TC-83.
4. The virus-like particle according to claim 1 , which comprises capsid, envelope protein E1, envelope protein E2 and envelope protein E3.
5. The virus-like particle according to claim 1 , which further comprises an envelope protein E2, and wherein the at least one antigen is further inserted into said envelope protein E2.
6. The virus-like particle according to claim 1 , wherein the at least one antigen is inserted between residues corresponding to residues 321 and 326 of SEQ ID NO: 1, residues 321 and 326 of SEQ ID NO: 2 or residues 330 and 335 of SEQ ID NO: 3.
7. The virus-like particle according to claim 1 , wherein the at least one antigen is an antigen from Plasmodium falciparum circumsporozoite protein, PD-1, PD-L1, CTLA-4, DISC1, IL-2, HER2, BTLA or HVEM.
8. The virus-like particle according to claim 7 , wherein a peptide selected from (1) and (2) is inserted into the envelope E3 protein:
(1) (NPNA)n (n=4-30) (SEQ ID NO. 140), and
(2) an amino acid sequence selected from SEQ ID NOs. 6-9 and 15-29.
9. The virus-like particle according to claim 1 , wherein the insertion of said at least one antigen into said furin cleavage site prevents cleavage of said site by furin.
10. An isolated nucleic acid molecule comprising a nucleotide sequence encoding the virus like particle according to claim 1 .
11. A vector comprising the nucleic acid molecule according to claim 10 , wherein the vector optionally comprises an expression control sequence operably linked to the nucleic acid molecule.
12. A composition comprising:
(a) the virus-like particle according to claim 1 ; and
(b) a pharmaceutically acceptable carrier.
13. A cell line expressing a viral structural protein which comprises an alternation/mutation to the amino acid sequence at the furin site in the envelope protein E3 according to claim 1 .
14. The cell line according to claim 13 , wherein the cell line is a stable cell line.
15. The cell line according to claim 13 , wherein the E3 is removed by a protease.
16. The virus-like particle according to claim 3 , wherein the viral structural protein is from one of SEQ ID NOs: 1-3.