The membrane-associated cytokines bound their corresponding surface receptors mainly in cis. Co-transfection of cytokine mRNAs with caTLR4 mRNA, caCD40 mRNA, or both, enhanced IFNγ production by T cells, induced upregulation of T cell activation molecules (CD25, 4-1BB, OX40), and improved the cytotoxicity of TILs against autologous melanoma cells in vitro in comparison to cells transfected with caTLR4 mRNA, caCD40 mRNA, or both [204, 205]. Here, IL2RA is linked to melanoma.