Viral particles encoding multifunctional cytokines
The present invention relates to a novel fusion protein with the formula X-Y, or Y-X, wherein X represents a first immunoregulating polypeptide and Y represents a second immunoregulating polypeptide different from X. The present invention also relates to a nucleic acid molecule encoding such a fusion protein and a vector comprising such a nucleic acid molecule. The present invention also provides infectious viral particles and host cells comprising such a nucleic acid molecule or such a vector as well as a process for producing such infectious viral particles. The present invention also relates to a method for recombinantly producing such a fusion protein. Finally, the present invention also provides a pharmaceutical composition comprising such a fusion protein, a nucleic acid molecule, a vector, infectious viral particles and a host cell as well as the therapeutic use thereof.
1. An isolated infectious viral particle comprising a polynucleotide encoding a fusion protein, wherein said fusion protein comprises the amino acid sequence of SEQ ID NO:1.
2. The isolated infectious viral particle of claim 1 , wherein:
said isolated infectious viral particle comprises a viral vector;
said viral vector comprises a polynucleotide encoding a fusion protein; and
said fusion protein comprises the amino acid sequence of SEQ ID NO:1.
3. The infectious viral particle of claim 2 , wherein said viral vector is selected from the group consisting of baculoviral vectors, papoviral vectors, herpes viral vectors, adenoviral vectors, adenovirus-associated viral vectors, poxviral vectors, foamy viral vectors, and retroviral vectors.
4. The isolated infectious viral particle of claim 3 , wherein said viral vector is an adenoviral vector.
5. The isolated infectious viral particle of claim 4 , wherein:
said adenoviral vector is an E1- and E3-deleted replication-defective adenoviral vector comprising a polynucleotide encoding a fusion protein inserted in replacement of the E1 region;
said fusion protein comprises the amino acid sequence of SEQ ID NO:1; and
said polynucleotide encoding a fusion protein is placed under the control of a CMV promoter.
6. The isolated infectious viral particle of claim 2 , wherein said viral vector further comprises one or more transgenes encoding (i) a tumor proliferation inhibitor and/or (ii) at least one antigen.
7. The isolated infectious viral particle of claim 6 , wherein said tumor proliferation inhibitor is a fusion protein encoding a two domain enzyme possessing both CDase and UPRTase activities.
8. The isolated infectious viral particle of claim 6 , wherein said antigen is a HPV antigen selected from the group consisting of E5, E6, E7, L1, L2, and combinations thereof.
9. The isolated infectious viral particle of claim 8 , wherein said HPV antigen is a membrane-anchored form of a non-oncogenic variant of the early HPV-16 E6 and/or E7 antigen.
10. An isolated host cell comprising the infectious viral particle of claim 1 .
11. An isolated host cell comprising the infectious viral particle of claim 2 .
12. A composition comprising the infectious viral particle of claim 1 and a pharmaceutically acceptable vehicle.
13. A composition comprising the infectious viral particle of claim 2 and a pharmaceutically acceptable vehicle.
14. A composition comprising the isolated host cell of claim 10 and a pharmaceutically acceptable vehicle.
15. A composition comprising the isolated host cell of claim 11 and a pharmaceutically acceptable vehicle.
16. A process for producing the infectious viral particle of claim 1 , comprising the steps of:
(a) introducing a viral vector into a suitable cell line, wherein said viral vector comprises a polynucleotide encoding a fusion protein, and said fusion protein comprises the amino acid sequence of SEQ ID NO:1;
(b) culturing said cell line under suitable conditions so as to allow the production of said infectious viral particle;
(c) recovering the produced infectious viral particle from the culture of said cell line; and
(d) optionally purifying said recovered infectious viral particle.