Presentation of mouse tum- P91A antigen from chimeric proteins with different subcellular localizations by class I molecules of the major histocompatibility complex.
Like many antigens presented by class I molecules the mouse Ld-binding tum- antigen P91A derives from a cytosolic protein. To decide how stringent this localization is for presentation to cytotoxic lymphocytes (CTL) the P91A template has been inserted in the cDNA of rat esterase ES-10, a protein located in the endoplasmic reticulum (ER), and in the cDNA of mouse interleukin-9, a secretory product of lymphocytes. The esterase construct was also engineered to replace the C-terminal leucine by arginine, which causes secretion of the protein, or to delete the N-terminal presequence, which prevents transfer of the nascent chain to the ER. After cell-free transcription-translation, or transfection in COS cells, the products of the chimeric cDNA had the expected size and localization; however, the truncated form of esterase remained undetected in COS cells. The various chimeric templates were transfected in P1.HTR cells (H-2d); upon challenge with Ld-restricted anti-P91A CTL the cells were lyzed almost as efficiently as cells transfected with the full-length P91A cDNA. We conclude that peptide fragments that bind to class I molecules of the major histocompatibility complex can be generated in the ER.
Godelaine
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D
Laboratoire de Chimie Physiologique, Université Catholique de Louvain, Brussels, Belgium.
Van Pel
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A
Van Malderen
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M
Beaufay
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H
D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology*; D000951:Antigens, Neoplasm / Q000276:immunology*; D001483:Base Sequence; D002451:Cell Compartmentation; D002522:Chlorocebus aethiops; D003602:Cytotoxicity, Immunologic; D006649:Histocompatibility Antigens / Q000276:immunology*; D015395:Histocompatibility Antigens Class I / Q000276:immunology*; D066298:In Vitro Techniques; D051379:Mice; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D011993:Recombinant Fusion Proteins / Q000276:immunology; D013347:Subcellular Fractions; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014162:Transfection
D000951:Antigens, Neoplasm; D006649:Histocompatibility Antigens; D015395:Histocompatibility Antigens Class I; D009838:Oligodeoxyribonucleotides; D011993:Recombinant Fusion Proteins; C070850:transplantation antigen P91A, mouse; C029237:tumor-associated transplantation antigen