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.
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.
Wroblewski
Joanne M
JM
Division of Endocrinology, Department of Internal Medicine, College of Medicine, University of Kentucky, Lexington, KY 40536-0093, USA.
Yannelli
John R
JR
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*
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