Related Experiment Video
Updated: Sep 28, 2026
Methodology for the Efficient Generation of Fluorescently Tagged Vaccinia Virus Proteins
Published on: January 17, 2014
[Antigen presentation efficiency of recombinant Vaccinia virus expressing minigene product OVA(257-264) in eukaryotic
1Department of Biochemistry, Laboratory of Molecular Biology, Shanghai Second Medical University, Shanghai 200025, China. qiangx@shsmu.edu.cn
Abstract:
rVV (recombinant vaccinia virus) expressing minigene products and rVV expressing full length OVA protein were used to infect several eukaryotic cells. Monoclonal antibody 25.D-1.16 (anti-OVA(257-264)/K(b)) was used to detect the conformation of H-2K(b)-OVA complex on the surface of these cells. The tetramer technique was used to determine their ability to induce antigen specific T cells. The results showed that the tetramer had been generated successfully. Moreover, L929-K(b) cells infected by rVV-OVA can be used as APC to induce CTL response. Specific CTL response was demonstrated by H-2K(b-OVA tetramer staining and cytotoxicity assay. Extracelluar IFN-gamma was also quantitatively analyzed. The results from H-2Kb)-OVA tetramer staining were consistent with cytotoxicity assay. It can be concluded that rVV expressing minigene product is a more efficient source of peptides for class I molecules, and can be more immunogenic than rVV expressing full length protein.
