Recombinant Vaccinia Virus MVA for Generation and Analysis of T Cell Responses Against Tumor Associated Antigens

Abstract

Live attenuated viruses used as vaccines are known for their efficacy to elicit protective immunity against viral diseases. More recently, with an increasing number of tumor-associated antigens (TAA) being identified and molecularly cloned (1) the development of vaccines for cancer immunotherapy has gained considerable interest. In particular, live recombinant viral vectors seem to be appropriate delivery systems for efficient presentation of TAA to the immune system. The promise of viral vectors is likely to be founded on their capacity for high-level expression of target genes combined with their intrinsic property to activate immunological control systems mimicking an infection with a disease causing agent.

Transient Host Range Selection for Genetic Engineering of Modified Vaccinia Virus Ankara

“…Genomic viral DNA was isolated from infected cells as described previously (5). Recombinant MVA were analyzed by PCR using oligonucleotides annealing within flanking regions of deletion III (MVA-III-5 ′ : 5 ′ -GAATGC -ACATACATAAGTACCGGCATCTCT -AGCAGT-3 ′ and MVA-III -3 ′ : 5 ′ -CAC -CAGCGTCTACATGACGAGCTTCC-GAGTTCC-3 ′ ), according to the following protocol: an initial step at 94°C for 2 min, followed by 30 cycles at 94°C for 30 s, 55°C for 40 s, 72°C for 3 min and a termination step at 72°C for 7 min.…”