title: In vitro generated Rh(null) red cells recapitulate the in vivo deficiency: a model for rare blood group phenotypes and erythroid membrane disorders.
abstract: Lentiviral modification combined with ex vivo erythroid differentiation was used to stably inhibit RhAG expression, a critical component of the Rh(rhesus) membrane complex defective in the Rh(null) syndrome. The cultured red cells generated recapitulate the major alterations of native Rh(null) cells regarding antigen expression, membrane deformability, and gas transport function, providing the proof of principle for their use as model of Rh(null) syndrome and to investigate Rh complex biogenesis in human primary erythroid cells. Using this model, we were able to reveal for the first time that RhAG extinction alone is sufficient to explain ICAM-4 and CD47 loss observed on native Rh(null) RBCs. Together with the effects of RhAG forced expression in Rh(null) progenitors, this strongly strengthens the hypothesis that RhAG is critical to Rh complex formation. The strategy is also promising for diagnosis purpose in order to overcome the supply from rare blood donors and is applicable to other erythroid defects and rare phenotypes, providing models to dissect membrane biogenesis of multicomplex proteins in erythroid cells, with potential clinical applications in transfusion medicine.
authors: Institut National de la Transfusion Sanguine (INTS), 75015, Paris, France.
authors: Marie
authors: M
authors: Cambot
authors: Christelle
authors: C
authors: Mazurier
authors: Florence
authors: F
authors: Canoui-Poitrine
authors: Nicolas
authors: N
authors: Hebert
authors: Julien
authors: J
authors: Picot
authors: Denis
authors: D
authors: Clay
authors: Véronique
authors: V
authors: Picard
authors: Pierre
authors: P
authors: Ripoche
authors: Luc
authors: L
authors: Douay
authors: Anne
authors: A
authors: Dubart-Kupperschmitt
authors: Jean-Pierre
authors: JP
authors: Cartron
mesh_terms: D053687:Adult Stem Cells / Q000166:cytology / Q000378:metabolism; D000745:Anemia, Hemolytic, Congenital / Q000378:metabolism / Q000473:pathology; D029502:Anemia, Hypoplastic, Congenital / Q000378:metabolism / Q000473:pathology; D001798:Blood Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D051928:CD47 Antigen / Q000378:metabolism*; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D002454:Cell Differentiation; D002460:Cell Line; D002478:Cells, Cultured; D041905:Erythroid Cells / Q000378:metabolism* / Q000473:pathology; D015672:Erythroid Precursor Cells / Q000166:cytology / Q000378:metabolism; D005260:Female; D005312:Fetal Blood; D053686:Fetal Stem Cells / Q000166:cytology / Q000378:metabolism; D030342:Genetic Diseases, Inborn / Q000097:blood / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D008562:Membrane Glycoproteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D017092:Porphyria, Erythropoietic / Q000378:metabolism / Q000473:pathology; D011247:Pregnancy; D034622:RNA Interference; D034741:RNA, Small Interfering; D012156:Reticulocytes / Q000378:metabolism / Q000473:pathology; D012204:Rh-Hr Blood-Group System / Q000097:blood / Q000378:metabolism*
chemical_list: D001798:Blood Proteins; D051928:CD47 Antigen; C497894:CD47 protein, human; D015815:Cell Adhesion Molecules; C095100:ICAM4 protein, human; D008562:Membrane Glycoproteins; C498523:RHAG protein, human; D034741:RNA, Small Interfering; D012204:Rh-Hr Blood-Group System; C026410:Rho(D) antigen