Cryptic CTL epitope on a murine sarcoma Meth A generated by exon extension as a novel mechanism.

Abstract

Using the recently developed ELISPOT cloning methodology, we obtained cDNA clone S35 coding for the Ag epitope recognized by a murine sarcoma Meth A-specific CTL clone AT-1. Analysis of truncated S35 constructs and overlapping peptides revealed that the peptide epitope was LGAEAIFRL. AT-1 CTL lysed peptide-pulsed CMS8 cells at a nanomolar concentration, and the peptide strongly stimulated IFN-gamma production in AT-1 CTL. Sequence homology indicated that the S35 was derived from a mouse homologue of human retinoic acid-regulated nuclear matrix-associated protein (ramp). The ramp gene consisted of 15 exons. The majority of the ramp mRNA was the transcript normally spliced between exons 14 and 15, but a minor population of mRNA with an extended exon 14 was also present in Meth A cells. The epitope was derived from the newly created open reading frame, which resulted from extension of exon 14 after splicing of the adjacent intronic sequence.

• Authors: Akiko Uenaka, Yoshiki Hirano, Hidenori Hata, Sanda Win, Toshiki Aji, Motoyuki Tanaka, Toshiro Ono, Jonathan C A Skipper, Kenji Shimizu, Eiichi Nakayama
• Journal: Journal of immunology (Baltimore, Md. : 1950) (J Immunol) Vol. 170 Issue 9 Pg. 4862-8 (May 01 2003) ISSN: 0022-1767 [Print] United States
• PMID: 12707369 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antigens, Neoplasm
• Carrier Proteins
• DNA-Binding Proteins
• Epitopes, T-Lymphocyte
• Transcription Factors
• ZMYM2 protein, human
• Zmym2 protein, mouse
• Methylcholanthrene
• Interferon-gamma

Topics

• Amino Acid Sequence
• Animals
• Antigens, Neoplasm (biosynthesis, genetics, isolation & purification, metabolism)
• Base Sequence
• Carrier Proteins (biosynthesis, genetics, isolation & purification, metabolism)
• Clone Cells
• Cloning, Molecular
• Cytotoxicity Tests, Immunologic
• DNA-Binding Proteins (biosynthesis, genetics, isolation & purification, metabolism)
• Enzyme-Linked Immunosorbent Assay
• Epitopes, T-Lymphocyte (biosynthesis, genetics, isolation & purification, metabolism)
• Exons (genetics, immunology)
• Gene Library
• Humans
• Interferon-gamma (analysis)
• Methylcholanthrene
• Mice
• Mice, Inbred BALB C
• Mice, Inbred DBA
• Molecular Sequence Data
• Sarcoma, Experimental (chemically induced, immunology)
• T-Lymphocytes, Cytotoxic (immunology)
• Transcription Factors