Evaluation of Measles Vaccine Virus as a Vector to Deliver Respiratory Syncytial Virus Fusion Protein or Epstein-Barr Virus Glycoprotein gp350

Mok, Hoyin; Cheng, Xing; Xu, Qi; Zengel, James R; Parhy, Bandita; Zhao, Jackie; Wang, C. Kathy; Jin, Hong

Generation of recombinant measles virus and vectored measles viruses. Rescue of the recombinant viruses from MV antigenomic cDNA was performed as previously described [16, 30, 31] . Briefly, HEp-2 cells were infected with MVA-T7 at a multiplicity of infectivity (moi) of 5 for an hour prior to transfection. A DNA mixture of pEZ (0.8 μg), pN (0.4 μg), pP (0.4 μg) and pL (0.2 μg) were cotransfected into MVA-T7 infected HEp-2 cells using Lipofectamine 2000 (Invitrogen, Carlsbad, CA). The culture medium was replaced with DMEM containing 2% FBS after overnight incubation at 37 o C. The transfected cells were incubated for an additional 3 to 4 days and the culture supernatant was used to infect fresh Vero cells to amplify the rescued virus. The rescued viruses were then plaque purified twice in Vero cells. The viruses rescued from pEZ, pEZ-RSVsF1, pEZ-RSVsF3, pEZ-EBVgp350 and pEZ-EBVgp350tr were designated as rEZ, sF1, sF3, gp350 and gp350tr, respectively.