[Antigen presentation efficiency of recombinant Vaccinia virus expressing minigene product OVA(257-264) in eukaryotic cells].
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.
Qian
Li
L
Department of Biochemistry, Laboratory of Molecular Biology, Shanghai Second Medical University, Shanghai 200025, China. qiangx@shsmu.edu.cn
Qian
Guan-Xiang
GX
D000818:Animals; D017951:Antigen Presentation / Q000276:immunology*; D002460:Cell Line; D003601:Cytotoxicity Tests, Immunologic; D004274:DNA, Recombinant / Q000235:genetics / Q000276:immunology; D005260:Female; D015870:Gene Expression; D006367:HeLa Cells; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D010047:Ovalbumin / Q000235:genetics / Q000276:immunology*; D014616:Vaccinia virus / Q000235:genetics*
D004274:DNA, Recombinant; D007371:Interferon-gamma; D010047:Ovalbumin