title: Recombinant transmembrane CD59 (CD59-TM) confers complement resistance to GPI-anchored protein defective melanoma cells.
abstract: Protectin (CD59) is a glycosylphosphatidylinositol (GPI)-anchored cell membrane glycoprotein, broadly expressed on melanocytic cells, that represents the main restriction factor of complement (C)-mediated lysis of human melanoma cells. Levels of CD59 expression may impair the clinical efficacy of C-activating monoclonal antibodies (mAb); thus, we investigated the molecular mechanisms underlying the lack of CD59 expression in selected melanoma cells. Serological and biochemical analyses showed that MeWo melanoma cells expressed CD59 neither at cell surface nor at cytoplasmic levels; however, no critical mutations were identified in their CD59 mRNA. Consistently, MeWo CD59 cDNA (MeWo-CD59) was appropriately translated when transfected into the CD59-positive Mel 100 melanoma cells, and into the CD59-negative Nalm-6 pre-B leukemia cells that acquired resistance to C. In contrast, transfection of MeWo cells with CD59 cDNA from Mel 275 melanoma cells did not induce CD59 expression; however, their transfection with the CD59-TM chimeric construct, obtained by replacing the GPI-anchoring signal of MeWo-CD59 with the transmembrane tail of the human low-density lipoprotein receptor, induced the expression of a C-protective transmembrane form of CD59. These data, together with the absent expression of additional GPI-anchored proteins (i.e., CD55), suggest that defects in the biosynthesis and/or processing of GPI-anchored proteins underlie the lack of CD59 expression in MeWo cells. Further unveiling of the molecular mechanism that turns off CD59 expression in human melanoma cells will help to set-up more effective therapeutic strategies utilizing C-activating mAb in melanoma patients.
authors.affiliation: Cancer Bioimmunotherapy Unit, Centro di Riferimento Oncologico, Istituto Nazionale di Ricovero e Cura a Carattere Scientifico, Aviano, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: De Nardo
authors.forename: Ester
authors.initials: E
authors.lastname: Fonsatti
authors.forename: Luca
authors.initials: L
authors.lastname: Sigalotti
authors.forename: Luana
authors.initials: L
authors.lastname: Calabrò
authors.forename: Francesca
authors.initials: F
authors.lastname: Colizzi
authors.forename: Enzo
authors.initials: E
authors.lastname: Cortini
authors.forename: Sandra
authors.initials: S
authors.lastname: Coral
authors.forename: Maresa
authors.initials: M
authors.lastname: Altomonte
authors.forename: Michele
authors.initials: M
authors.lastname: Maio
mesh_terms: D018957:CD59 Antigens / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D003165:Complement System Proteins / Q000494:pharmacology / Q000502:physiology*; D004351:Drug Resistance / Q000502:physiology; D015870:Gene Expression; D017261:Glycosylphosphatidylinositols / Q000172:deficiency*; D006801:Humans; D008545:Melanoma / Q000503:physiopathology*; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D011994:Recombinant Proteins / Q000378:metabolism; D014407:Tumor Cells, Cultured
chemical_list: D018957:CD59 Antigens; D017261:Glycosylphosphatidylinositols; D020033:Protein Isoforms; D011994:Recombinant Proteins; D003165:Complement System Proteins