SEREX analysis for tumor antigen identification in a mouse model of adenocarcinoma

Abstract

Evaluation of immunotherapy strategies in mouse models of carcinoma is hampered by the limited number of known murine tumor antigens (Ags). Although tumor Ags can be identified based on cytotoxic T-cell activation, this approach is not readily accomplished for many tumor types. We applied an alternative strategy based on a humoral immune response, SEREX, to the identification of tumor Ags in the murine colon adenocarcinoma cell line MC38. Immunization of syngeneic C57BL/6 mice with MC38 cells by three different methods induced a protective immune response with concomitant production of anti-MC38 antibodies. Immunoscreening of an MC38-derived expression library resulted in the identification of the endogenous ecotropic leukemia virus envelope (env) protein and the murine ATRX protein as candidate tumor Ags. Northern blot analysis demonstrated high levels of expression of the env transcript in MC38 cells and in several other murine tumor cell lines, whereas expression in normal colonic epithelium was absent. ATRX was found to be variably expressed in tumor cell lines and in normal tissue. Further analysis of the expressed env sequence indicated that it represents a nonmutated tumor Ag. Polynucleotide immunization with DNA encoding the env polypeptide resulted in strong and specific antibody responses to this self Ag in all immunized mice. Thus, SEREX offers a rapid means of identifying tumor Ags in murine cancer models. Cancer Gene Therapy (2000) 7, 446 -455

Identification of a novel prostate cancer‐associated tumor antigen

“…Following the initial development of SEREX by Sahin et al [9] the serological approach for the identification of human tumor antigens has been applied to several tumor types [11][12][13][14]20,23,27,[30][31][32][33][34][35][36][37][38][39][40] resulting in the identification of more than 2,500 antigens, one-third of which are novel. A proportion of tumor antigens, identified via this method and other approaches, form a unique class of differentiation antigens with expression confined to tumor tissue and normal testis, placenta and embryonic tissue; this led to their designation as cancer testis antigens [18,25,26,[41][42][43][44][45][46].…”

PHF3-specific antibody responses in over 60% of patients with glioblastoma multiforme

“…Bearing in mind these assumptions, it is important to focus the search for GBM-related antigens on those antigens that have induced an antibody response in the patient. We used a combined immunological and molecular approach known as the SEREX method to detect GBM-related antigens (Sahin et al, 1995;Tureci et al, 1997;Hampton et al, 2000). PHF3 was identi®ed from a GBM cDNA library using autologous patient serum in the immunological screening.…”

Molecular effects of dADD1 misexpression in chromatin organization and transcription

“…There have been studies linking different levels of expression of ATRX as drivers of specific phenotypes that give rise to disease and cancer [13,18,19]. In the present study, we searched human somatic cancer databases and found that the ATRX gene is overexpressed in a wide variety of human cancers.…”