Analysis of Tumor Rejection Antigen of Murine Myeloma MOPC70A Recognized by Specific Cytotoxic T cell.

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Murine myeloma MOPC70A specific cytotoxic T cell (CTL) clone E10 was established from spleen cells of tumor bearing BALB/c mouse. 2. Expression cDNA library was cloned from mRNA of MOPC70A into mammalian expression vector pMET7 and divided into pools of 100 bacterial colonies. And the total 56,000 cDNA clones were tested for their ability to stimulate TNFα production by E10 CTL after transfection into COS7 transfected H-2 gene. 3. A clone (2D2) which stimulate E10 to produce TNFα and IFNγ was cloned. 4. The sequence of 2D2 provided 1398 bp long. Sequence homology analysis revealed that 2D2 is murine apoptosis inducing factor (mAIF). 5. To identify antigenic CTL epitope encoded by the 2D2 gene, the ability of deletion mutant of 2D2 gene to stimulate E10 to produce TNFα were investigated. The results suggested that the antigenic epitope resided within the region of 737-870bp of 2D2. From analysis of the synthetic candidate peptide on the region of 737-870 bp, a nonamer peptide DLGPDVGYEA sensitized E10 CTL was found as an epitope.