title: Identification of HLA-CW3, GNAS and IMPA as cytotoxic T-lymphocyte (CTL) target antigens using an allogeneic mixed lymphocyte tumor cell culture (MLTC) system and subsequent cDNA library screening.
abstract: Allogeneic mixed lymphocyte tumor cell cultures (MLTCs) were established using lymphocytes from non-small-cell lung cancer (NSCLC) patient UKY-53 and HLA-A2+ NSCLC tumor cells (UKY-29). The tumor cells expressed the lymphocyte costimulatory molecule CD80 (UKY29.7). Cytolytic activity showed the cytotoxic T-lymphocytes (CTL) lysed UKY-29, but not K562 or Daudi. The CTL also lysed: HLA-A2+ and -A24+ tumor cell lines from a number of tumor histologies. The CTL also lysed Epstein Barr virus transformed (EBV) B-cells, UKY-29EBV, autologous to the stimulating cell line, UKY29TC. These data suggested the presence of both tumor-specific and allogeneic reactivities in the bulk CTL population. Subsequent cDNA cloning analysis and sequencing demonstrated that the bulk CTL population was recognizing: (i) allogeneic target HLA-CW3, and two minor histocompatibility antigens; (ii) guanine nucleotide-binding protein, G(S) (GNAS), and (iii) inositol myophosphatase (IMPA). All three antigens, we believe, were restricted by HLA-A2. Whereas the system described was initially intended to identify tumor-associated antigens recognized by CTL, the nature of the allogeneic system provides a unique opportunity for the identification of epitopes that confer both allo and minor antigen recognition.
authors.affiliation: Division of Endocrinology, Department of Internal Medicine, College of Medicine, University of Kentucky, Lexington, KY 40536-0093, USA.
authors.forename: Joanne M
authors.initials: JM
authors.lastname: Wroblewski
authors.forename: John R
authors.initials: JR
authors.lastname: Yannelli
mesh_terms: D001483:Base Sequence; D045744:Cell Line, Tumor; D002864:Chromogranins; D018076:DNA, Complementary / Q000235:genetics; D019205:GTP-Binding Protein alpha Subunits, Gs / Q000235:genetics / Q000276:immunology*; D019204:GTP-Binding Proteins / Q000235:genetics / Q000276:immunology*; D015723:Gene Library; D015789:HLA-A2 Antigen / Q000276:immunology; D015236:HLA-C Antigens / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007959:Lymphocyte Culture Test, Mixed; D008969:Molecular Sequence Data; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics / Q000276:immunology*; D012333:RNA, Messenger / Q000235:genetics; D012680:Sensitivity and Specificity; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology* / Q000378:metabolism*
chemical_list: D002864:Chromogranins; D018076:DNA, Complementary; D015789:HLA-A2 Antigen; D015236:HLA-C Antigens; C038653:HLA-C*03 antigen; D012333:RNA, Messenger; D010744:Phosphoric Monoester Hydrolases; C406732:GNAS protein, human; D019204:GTP-Binding Proteins; D019205:GTP-Binding Protein alpha Subunits, Gs