| Vector | adenovirus |
| Construct | Ad5/35-E2F-delta24 |
| Vector type | novel oncolytic serotype 5/35 adenovirus |
| Transgene/Inserted gene | CD40L, which includes CD40-mediated tumor cell apoptosis and adaptive immune activation; and 4-1BBL, which enhances immunologic memory and expands natural killer (NK) cells. |
| Viral genome modification | The gene sequence of trimerized membrane-bound (TMZ)-CD40L, alone or in combination with 4-1BBL (LOAd703), separated by a T2A peptide, was synthesized with a cytomegalovirus (CMV) promoter upstream of the 5′ gene and adenovirus-flanking regions for homologous recombination into adenoviral vectors at both ends. |
| Vector production method | Viruses were produced by transfection of 293 cells (ATCC), followed by expansion in A549 cells (ATCC). The virus supernatants were purified by CsCl gradient centrifugation and diluted in 20 mmol/L Tris, 25 mmol/L NaCl, and 2.5% glycerol. The viable virus titer was measured using a fluorescence-forming units (ffu) assay. The LOAd(−) is the basic LOAd virus without the CMV transgene cassette. |
| Additional feature | CD40L|4-1BBL |