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. 2005 Aug 19;94(11):931–936. doi: 10.1111/j.1349-7006.2003.tb01380.x

Murine leukemia RL1 and sarcoma Meth A antigens recognized by cytotoxic T lymphocytes (CTL)

Akiko Uenaka 1,✉, Eiichi Nakayama 1
PMCID: PMC11160269  PMID: 14611667

Abstract

Peptide elution and expression cloning methods have been used to identify T cell‐recognized antigens for which no molecular information is available. We identified a unique tumor antigen peptide pRL1a, IPGLPLSL that is recognized by CTL on BALB/c RLInline graphic1 leukemia by peptide elution. The sequence of the peptide corresponded to the normally untranslated 5’region of akt. Cytotoxicity was generated in BALB/c spleen cells by in vivo and in vitro sensitization with pRL1a peptide in the form of multiple antigen peptide (MAP), but not the original form. pRL1a MAP immunization had a significant growth‐inhibitory effect. pRL1a MAP was mostly internalized into the endosomal compartment of antigen‐presenting cells, leaked to the cytosol, and degraded, and the pRL1a peptide produced was presented through the MHC class I pathway. In vivo depletion of CD4 T cells from tumor‐inoculated BALB/c mice caused RLInline graphic1 regression. Overexpression of the RLakt molecule seemed to induce CD4 immunoregulatory cells, which resulted in progressive RLInline graphic1 growth in BALB/c mice. In vivo administration of anti‐CD25 mAb (PC61) caused regression of RLInline graphic1, suggesting that CD4+CD25+ immunoregulatory cells were involved in the tumor growth. Recently, we improved the sensitivity and the efficacy of T cell antigen cloning from cDNA expression libraries by using large‐ and small‐scale ELISPOT assays. Using the IFN‐γ ELISPOT method, we obtained a cDNA clone S35 of 937 bp recognized by AT‐1 CTL on BALB/c Meth A sarcoma. S35 was a part of the retinoic acid‐regulated nuclear matrix‐associated protein (ramp). AT‐1 CTL recognized the peptide LGAE‐AIFRL, which was derived from a newly created open reading frame due to the exon 14 extension.

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