In vivo CTL Immunity

In vivo CTL Immunity
Can be Elicited by OVA-linker-β2m Fusion Protein

QIAN Li, QIAN Guan-Xiang*
( Research Centre of Molecular Biology, Shanghai Second Medical University,
Shanghai 200025, China )

Abstract    To study
the effect of OVA-linker-β2m fusion protein as an immunogen to induce CTL
mediated immune response in mice. The β2m gene was amplified by PCR. OVA257-264 peptide sequence and an
8-amino acid linker were added to the COOH-terminus of β2m gene, and then
cloned into pE42b(+) vector. After that, gene expression, protein purification
and refolding were conducted. Then the protein product was used as an immunogen
to induce an efficient CTL mediated immune response in vivo.
Specific CTL responses were demonstrated by H-2K^b-OVA tetramer
staining and cytotoxicity assay. Extracelluar IFN-γ was also quantitatively
analyzed. The results showed that specific CTL response could be induced in
vivo by OVA-linker-β2m fusion protein. Generation of terameric
peptide-MHC complexes in vitro is a powerful tool to
stain specific CTL.

Key words tetramer; gene expression; purification； refolding; CTL

^*Corresponding authors:  Tel, 86-21-63846590-776881; Fax,
86-21-63842916; e-mail, [email protected]