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The Journal of Immunology, Vol 154, Issue 9 4630-4641, Copyright © 1995 by American Association of Immunologists

ARTICLES

Multiple point mutations in an endogenous retroviral gene confer high immunogenicity on a drug-treated murine tumor

U Grohmann, P Puccetti, ML Belladonna, F Fallarino, R Bianchi, L Binaglia, K Sagakuchi, MG Mage, E Appella and MC Fioretti
Department of Experimental Medicine and Biochemical Sciences, University of Perugia, Italy.

Exposure in vivo of murine L5178Y lymphoma cells to cytoreductive triazene derivatives leads to the generation of immunogenic variant lines expressing new transplantation Ags recognized by CTL. In one such clonal variant (clone D), at least one subset of T cell neoepitopes are provided by proteins previously shown by serology to be products of endogenous retroviral env sequences. We report here on characterization of PCR-amplified gp70 env genes in clone D. Relative to known gp70 sequences in parental cells and in current databases, one gp70 sequence presented four distinct nucleotide changes, two of which were apparently unique to clone D DNA and cDNA upon differential hybridization analysis. Transfection experiments with the entire gp70 gene or subgenic fragments encompassing a single putative mutation showed that products of the mutated env gene or fragments may confer immunogenicity in vivo and susceptibility in vitro to lysis by clone D- primed, H-2Kd- or H-2Ld-restricted CTL. By skin test assay of mice primed with either clone D or three mutated synthetic peptides, evidence was obtained that amino acid substitutions at the relevant positions of the gp70 protein may produce immunogenic T cell epitopes and that these epitopes are expressed in vivo by clone D. These data point to the role of mutated retroviral tumor peptides as rejection Ags in histocompatible hosts.


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