title: Interferon-gamma rescues HLA class Ia cell surface expression in term villous trophoblast cells by inducing synthesis of TAP proteins.
abstract: Human placental trophoblast cells that constitute the materno-fetal interface during pregnancy escape maternal alloimmune attack. The different trophoblast cell subpopulations have developed efficient regulatory mechanisms to prevent expression of beta2-microglobulin-associated HLA class Ia molecules at their cell surface. We previously reported the presence of HLA class Ia messages in villous cytotrophoblast cells and in the syncytiotrophoblast differentiated in vitro purified from term placenta. In this study, we found that these transcripts are translated in heavy chain proteins that are endoglycosidase H sensitive and thus retained in the endoplasmic reticulum or cis-Golgi. Moreover, these class Ia heavy chains can be co-immunoprecipitated with the chaperone protein calnexin resident in the endoplasmic reticulum. When these trophoblast cells are treated with interferon (IFN)-gamma, part of the class Ia heavy chains become endoglycosidase H resistant, demonstrating that they have left the endoplasmic reticulum. Furthermore, after such a treatment, these heavy chains are detectable at the cell surface of these trophoblast cells, as assessed by two-color flow cytometry analysis and immunoprecipitation of cell surface biotinylated proteins using the W6/32 anti-HLA class I monoclonal antibody (mAb). IFN-gamma treatment induces a significant enhancement of the transcription of transporters associated with antigen processing (TAP1 and TAP2) rather than an increase of HLA class I or beta2-microglobulin messages. Finally, we demonstrate that an anti-TAP1 mAb co-immunoprecipitates TAP1 proteins and HLA class Ia heavy chains in these IFN-gamma-treated trophoblast cells. Thus, the constitutive absence of HLA class Ia cell surface expression in term villous cytotrophoblast and syncytiotrophoblast is likely to be due to a lack of transporter proteins that participate in the proper assembly of these molecules in the endoplasmic reticulum. Such a defect can be modified upon IFN-gamma treatment.
authors.affiliation: INSERM U395, Centre Hospitalo-Universitaire Purpan, Toulouse, France.
authors.forename: A M
authors.initials: AM
authors.lastname: Rodriguez
authors.forename: V
authors.initials: V
authors.lastname: Mallet
authors.forename: F
authors.initials: F
authors.lastname: Lenfant
authors.forename: J
authors.initials: J
authors.lastname: Arnaud
authors.forename: M
authors.initials: M
authors.lastname: Girr
authors.forename: S
authors.initials: S
authors.lastname: Urlinger
authors.forename: A
authors.initials: A
authors.lastname: Bensussan
authors.forename: P
authors.initials: P
authors.lastname: Le Bouteiller
mesh_terms: D000071181:ATP Binding Cassette Transporter, Subfamily B, Member 2; D000071450:ATP Binding Cassette Transporter, Subfamily B, Member 3; D018528:ATP-Binding Cassette Transporters / Q000096:biosynthesis*; D000911:Antibodies, Monoclonal; D002135:Calcium-Binding Proteins / Q000378:metabolism; D037281:Calnexin; D002470:Cell Survival; D002478:Cells, Cultured; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005260:Female; D006680:HLA Antigens / Q000378:metabolism; D059951:HLA-G Antigens; D006596:Hexosaminidases / Q000494:pharmacology; D015395:Histocompatibility Antigens Class I / Q000378:metabolism*; D006801:Humans; D016130:Immunophenotyping; D007371:Interferon-gamma / Q000494:pharmacology*; D008198:Lymph Nodes / Q000166:cytology*; D010920:Placenta / Q000166:cytology*; D011247:Pregnancy; D014176:Protein Biosynthesis; D011994:Recombinant Proteins; D013950:Thymus Gland / Q000166:cytology*; D014158:Transcription, Genetic; D014327:Trophoblasts / Q000166:cytology / Q000276:immunology*
chemical_list: D000071181:ATP Binding Cassette Transporter, Subfamily B, Member 2; D000071450:ATP Binding Cassette Transporter, Subfamily B, Member 3; D018528:ATP-Binding Cassette Transporters; D000911:Antibodies, Monoclonal; D002135:Calcium-Binding Proteins; D006680:HLA Antigens; D059951:HLA-G Antigens; D015395:Histocompatibility Antigens Class I; D011994:Recombinant Proteins; C073119:TAP1 protein, human; D037281:Calnexin; C071692:TAP2 protein, human; D007371:Interferon-gamma; D006596:Hexosaminidases