Enhanced antigen presenting ability of RNA CAR T cells by co-introduction of costimulatory molecules

A method of generating a T cell having enhanced anti-tumor activity, the method comprising introducing a nucleic acid in a T cell, wherein:

• (a) the nucleic acid comprises a first nucleic acid sequence encoding a chimeric antigen receptor (CAR) comprising an extracellular domain, a transmembrane domain, and an intracellular signaling domain, wherein the intracellular signaling domain comprises a costimulatory signaling domain that is a functional signaling domain from a protein selected from the group consisting of OX40, CD27, CD28, CD30, CD40, PD-1, CD2, CD7, CD258, NKG2C, B7-H3, ICAM-1, LFA-1 (CD11a/CD18), ICOS and 4-1BB (CD137), or any combination thereof, and
• (b) the nucleic acid comprises a second nucleic acid sequence encoding a polypeptide which enhances T cell priming, or a functional fragment or variant thereof, wherein the polypeptide is selected from the group consisting of CD70, CD86, and CD252,
• wherein the polypeptide is expressed at a level that:
• (1) the T cell enhances the priming of another T cell that does not comprise the nucleic acid and (2) expression of the polypeptide does not inhibit the cell-killing function of the CAR by more than 40%,
• provided that
• (i) the first and/or second nucleic acid sequence comprises an RNA;
• or
• (ii) the CAR further comprises a second intracellular signaling domain.