Defective glycosylation and multisystem abnormalities characterize the primary immunodeficiency XMEN disease
Juan C. Ravell, … , Matthias Mann, Michael J. Lenardo
Juan C. Ravell, … , Matthias Mann, Michael J. Lenardo
Published November 5, 2019
Citation Information: J Clin Invest. 2020;130(1):507-522. https://doi.org/10.1172/JCI131116.
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Research Article Immunology

Defective glycosylation and multisystem abnormalities characterize the primary immunodeficiency XMEN disease

Abstract

X-linked immunodeficiency with magnesium defect, EBV infection, and neoplasia (XMEN) disease are caused by deficiency of the magnesium transporter 1 (MAGT1) gene. We studied 23 patients with XMEN, 8 of whom were EBV naive. We observed lymphadenopathy (LAD), cytopenias, liver disease, cavum septum pellucidum (CSP), and increased CD4–CD8–B220–TCRαβ+ T cells (αβDNTs), in addition to the previously described features of an inverted CD4/CD8 ratio, CD4+ T lymphocytopenia, increased B cells, dysgammaglobulinemia, and decreased expression of the natural killer group 2, member D (NKG2D) receptor. EBV-associated B cell malignancies occurred frequently in EBV-infected patients. We studied patients with XMEN and patients with autoimmune lymphoproliferative syndrome (ALPS) by deep immunophenotyping (32 immune markers) using time-of-flight mass cytometry (CyTOF). Our analysis revealed that the abundance of 2 populations of naive B cells (CD20+CD27–CD22+IgM+HLA-DR+CXCR5+CXCR4++CD10+CD38+ and CD20+CD27–CD22+IgM+HLA-DR+CXCR5+CXCR4+CD10–CD38–) could differentially classify XMEN, ALPS, and healthy individuals. We also performed glycoproteomics analysis on T lymphocytes and show that XMEN disease is a congenital disorder of glycosylation that affects a restricted subset of glycoproteins. Transfection of MAGT1 mRNA enabled us to rescue proteins with defective glycosylation. Together, these data provide new clinical and pathophysiological foundations with important ramifications for the diagnosis and treatment of XMEN disease.

Authors

Juan C. Ravell, Mami Matsuda-Lennikov, Samuel D. Chauvin, Juan Zou, Matthew Biancalana, Sally J. Deeb, Susan Price, Helen C. Su, Giulia Notarangelo, Ping Jiang, Aaron Morawski, Chrysi Kanellopoulou, Kyle Binder, Ratnadeep Mukherjee, James T. Anibal, Brian Sellers, Lixin Zheng, Tingyan He, Alex B. George, Stefania Pittaluga, Astin Powers, David E. Kleiner, Devika Kapuria, Marc Ghany, Sally Hunsberger, Jeffrey I. Cohen, Gulbu Uzel, Jenna Bergerson, Lynne Wolfe, Camilo Toro, William Gahl, Les R. Folio, Helen Matthews, Pam Angelus, Ivan K. Chinn, Jordan S. Orange, Claudia M. Trujillo-Vargas, Jose Luis Franco, Julio Orrego-Arango, Sebastian Gutiérrez-Hincapié, Niraj Chandrakant Patel, Kimiyo Raymond, Turkan Patiroglu, Ekrem Unal, Musa Karakukcu, Alexandre G.R. Day, Pankaj Mehta, Evan Masutani, Suk S. De Ravin, Harry L. Malech, Grégoire Altan-Bonnet, V. Koneti Rao, Matthias Mann, Michael J. Lenardo

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Figure 5

Selective N-glycosylation defects in XMEN disease.

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Selective N-glycosylation defects in XMEN disease. (A) Model of MAGT1 as...
(A) Model of MAGT1 as a facilitator of STT3B-dependent transference of oligosaccharides (blue and pale pink) to glycosites (red) of nascent peptides (gray) in the ER (13, 15, 16, 19). (B) Flow cytometric histogram and MFI quantification relative to HCs of NKG2D and CD5 in CD8+T cells from HCs (blue) and patients with XMEN (red), with an unstained control (gray, n = 6). (C) Immunoblot of NKG2D, MAGT1, and β-tubulin in T cells from HCs and patients with XMEN, with (+) or without (–) PNGase F treatment. Unglycosylated (0), partially glycosylated (1), and fully glycosylated (3) NKG2D bands and nonspecific (2) and PNGase F (*) bands. (D) Flow cytometric histogram and MFI quantification relative to HCs of NKG2D and CD5 in CD8+ T cells treated for 24 hours with DMSO vehicle (blue) or tunicamycin (red) with unstained control (gray). PE, fluorophore R-phycoerythrin. (E) Immunoblot of NKG2D and β-tubulin in T cells with (+) or without (–) PNGase F treatment after incubation with DMSO (Veh) or 10 μg/mL tunicamycin (Tun) for 18 hours. Fully glycosylated (1) and unglycosylated (0) NKG2D and PNGase F (*) bands. (F) Heatmap depicting significantly different glycopeptides between HCs (n = 3) and patients with XMEN (n = 3), with increased (red) and decreased (black) glycosylation shown. Numbers on the left for immunoblots indicate kDa standards. Data are representative of 6 (C) and 3 (E) replicates, respectively. Data represent the mean ± SEM. ****P < 0.0001, by 1-sample t test with μ = 1 (B and D) and 2-sample t test (F).