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 nucleic acid molecule encoding a fusion protein comprising the amino acid sequence of SEQ ID NO: 1.
2. An isolated vector comprising the nucleic acid molecule of claim 1 .
3. The isolated vector of claim 2 , wherein said vector is a bacterial plasmid, a bacteriophage vector, a yeast episomal vector, an artificial chromosomal vector, or a viral vector.
4. The isolated vector of claim 3 , wherein said vector is a viral vector.
5. The isolated vector of claim 4 , 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.
6. The isolated vector of claim 5 , wherein said vector is an adenoviral vector.
7. The isolated vector of claim 6 , wherein said vector is an E1- and E3-deleted replication-defective adenoviral vector comprising an isolated nucleic acid molecule encoding a fusion protein comprising the amino acid sequence of SEQ ID NO: 1 inserted in replacement of the E1 region and placed under the control of the CMV promoter.
8. The isolated vector of claim 2 , wherein said vector further comprises one or more transgenes encoding (i) a tumor proliferation inhibitor and/or (ii) at least one antigen.
9. The isolated vector of claim 8 , wherein said tumor proliferation inhibitor is a fusion protein which encodes a two domain enzyme possessing both CDase and UPRTase activities.
10. The isolated vector of claim 8 , wherein said antigen is a HPV antigen selected from the group consisting of E5, E6, E7, L1, and L2 either individually or in combination.
11. The isolated vector of claim 10 , wherein said HPV antigen is a Membrane-anchored form of a non-oncogenic variant of early HPV-16 E6 and/or E7 antigen.
12. An isolated host cell comprising the nucleic acid molecule of claim 1 .
13. An isolated host cell comprising the vector of claim 2 .
14. A composition comprising the vector of claim 2 and a pharmaceutically acceptable vehicle.
15. A composition comprising the host cell of claim 12 and a pharmaceutically acceptable vehicle.
16. A composition comprising the host cell of claim 13 and a pharmaceutically acceptable vehicle.
17. A method for producing a fusion protein, comprising the steps of:
(a) introducing the vector of claim 2 into a suitable host cell to produce a transfected or infected host cell,
(b) culturing in-vitro said transfected or infected host cell under conditions suitable for growth of the host cell,
(c) recovering said fusion protein from the culture of said host cell, and
(d) optionally purifying said recovered fusion protein.