Engineered cells expressing multiple immunomodulators and uses thereof

An in vitro engineered immune cell comprising an adenoviral vector for conditionally expressing proteins, the vector comprising a polynucleotide encoding an ecdysone gene switch, wherein said polynucleotide comprises

• (1) a polynucleotide sequence comprising a first transcription factor sequence and a second transcription factor sequence under the control of a promoter, wherein the proteins encoded by the first transcription factor sequence and second transcription factor sequence interact to form a ligand-dependent transcription factor complex,
• wherein the first transcription factor sequence comprises a nucleic acid encoding a VP-16 transactivation domain and a retinoic acid-X-receptor (RXR) polypeptide,
• wherein the RXR polypeptide is a genetically engineered chimera comprising vertebrate RXR and invertebrate RXR domains,
• wherein said vertebrate RXR ligand binding domain is a human RXR ligand binding domain,
• wherein the second transcription factor sequence comprises a nucleic acid encoding a GAL-4 DNA binding domain and an ecdysone receptor protein,
• wherein said ecdysone receptor ligand binding domain is derived from a Choristoneura fumiferna ecdysone receptor ligand binding domain, and
• wherein said Choristoneura fumiferna ecdysone receptor ligand binding domain further comprises a genetically engineered substitution mutation compared to the naturally occurring Choristoneura fumiferna ecdysone receptor ligand binding domain from which it was derived;
• (2) a polynucleotide encoding a polypeptide at least 90% identical to IL-12;
• and
• (3) a polynucleotide encoding a polypeptide at least 90% identical to IL-21;
• wherein at least one polynucleotide of (2) or (3) are linked to a promoter capable of being activated by said ligand dependent transcription factor complex.