title: Rapid T cell-based identification of human tumor tissue antigens by automated two-dimensional protein fractionation.
abstract: Identifying the antigens that have the potential to trigger endogenous antitumor responses in an individual cancer patient is likely to enhance the efficacy of cancer immunotherapy, but current methodologies do not efficiently identify such antigens. This study describes what we believe to be a new method of comprehensively identifying candidate tissue antigens that spontaneously cause T cell responses in disease situations. We used the newly developed automated, two-dimensional chromatography system PF2D to fractionate the proteome of human tumor tissues and tested protein fractions for recognition by preexisting tumor-specific CD4+ Th cells and CTLs. Applying this method using mice transgenic for a TCR that recognizes an OVA peptide presented by MHC class I, we demonstrated efficient separation, processing, and cross-presentation to CD8+ T cells by DCs of OVA expressed by the OVA-transfected mouse lymphoma RMA-OVA. Applying this method to human tumor tissues, we identified MUC1 and EGFR as tumor-associated antigens selectively recognized by T cells in patients with head and neck cancer. Finally, in an exemplary patient with a malignant brain tumor, we detected CD4+ and CD8+ T cell responses against two novel antigens, transthyretin and calgranulin B/S100A9, which were expressed in tumor and endothelial cells. The immunogenicity of these antigens was confirmed in 4 of 10 other brain tumor patients. This fast and inexpensive method therefore appears suitable for identifying candidate T cell antigens in various disease situations, such as autoimmune and malignant diseases, without being restricted to expression by a certain cell type or HLA allele.
authors.affiliation: Translational Immunology Unit, German Cancer Research Center, Heidelberg, Germany.
authors.forename: Philipp
authors.initials: P
authors.lastname: Beckhove
authors.forename: Rolf
authors.initials: R
authors.lastname: Warta
authors.forename: Britt
authors.initials: B
authors.lastname: Lemke
authors.forename: Diana
authors.initials: D
authors.lastname: Stoycheva
authors.forename: Frank
authors.initials: F
authors.lastname: Momburg
authors.forename: Martina
authors.initials: M
authors.lastname: Schnölzer
authors.forename: Uwe
authors.initials: U
authors.lastname: Warnken
authors.forename: Hubertus
authors.initials: H
authors.lastname: Schmitz-Winnenthal
authors.forename: Rezvan
authors.initials: R
authors.lastname: Ahmadi
authors.forename: Gerhard
authors.initials: G
authors.lastname: Dyckhoff
authors.forename: Mariana
authors.initials: M
authors.lastname: Bucur
authors.forename: Simone
authors.initials: S
authors.lastname: Jünger
authors.forename: Thomas
authors.initials: T
authors.lastname: Schueler
authors.forename: Volker
authors.initials: V
authors.lastname: Lennerz
authors.forename: Thomas
authors.initials: T
authors.lastname: Woelfel
authors.forename: Andreas
authors.initials: A
authors.lastname: Unterberg
authors.forename: Christel
authors.initials: C
authors.lastname: Herold-Mende
mesh_terms: D000818:Animals; D000941:Antigens / Q000276:immunology / Q000378:metabolism; D000951:Antigens, Neoplasm / Q000737:chemistry* / Q000276:immunology* / Q000378:metabolism; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D045142:Cross-Priming / Q000276:immunology; D006680:HLA Antigens / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007167:Immunotherapy / Q000379:methods; D051379:Mice; D008810:Mice, Inbred C57BL; D009369:Neoplasms / Q000276:immunology / Q000378:metabolism; D010455:Peptides / Q000737:chemistry* / Q000276:immunology* / Q000378:metabolism; D011506:Proteins / Q000276:immunology / Q000378:metabolism; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology / Q000378:metabolism
chemical_list: D000941:Antigens; D000951:Antigens, Neoplasm; D006680:HLA Antigens; D010455:Peptides; D011506:Proteins
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