title: Genetically engineered human islets protected from CD8-mediated autoimmune destruction in vivo.
abstract: Islet transplantation is a promising therapy for type 1 diabetes, but graft function and survival are compromised by recurrent islet autoimmunity. Immunoprotection of islets will be required to improve clinical outcome. We engineered human β cells to express herpesvirus-encoded immune-evasion proteins, "immunevasins." The capacity of immunevasins to protect β cells from autoreactive T-cell killing was evaluated in vitro and in vivo in humanized mice. Lentiviral vectors were used for efficient genetic modification of primary human β cells without impairing their function. Using a novel β-cell-specific reporter gene assay, we show that autoreactive cytotoxic CD8(+) T-cell clones isolated from patients with recent onset diabetes selectively destroyed human β cells, and that coexpression of the human cytomegalovirus-encoded US2 protein and serine proteinase inhibitor 9 offers highly efficient protection in vitro. Moreover, coimplantation of these genetically modified pseudoislets with β-cell-specific cytotoxic T cells into immunodeficient mice achieves preserved human insulin production and C-peptide secretion. Collectively, our data provide proof of concept that human β cells can be efficiently genetically modified to provide protection from killing mediated by autoreactive T cells and retain their function in vitro and in vivo.
authors.affiliation: Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, The Netherlands. A.Zaldumbide@lumc.nl
authors.forename: Arnaud
authors.initials: A
authors.lastname: Zaldumbide
authors.forename: Gonnie
authors.initials: G
authors.lastname: Alkemade
authors.forename: Françoise
authors.initials: F
authors.lastname: Carlotti
authors.forename: Tatjana
authors.initials: T
authors.lastname: Nikolic
authors.forename: Joana Rf
authors.initials: JR
authors.lastname: Abreu
authors.forename: Marten A
authors.initials: MA
authors.lastname: Engelse
authors.forename: Anja
authors.initials: A
authors.lastname: Skowera
authors.forename: Eelco J
authors.initials: EJ
authors.lastname: de Koning
authors.forename: Mark
authors.initials: M
authors.lastname: Peakman
authors.forename: Bart O
authors.initials: BO
authors.lastname: Roep
authors.forename: Rob C
authors.initials: RC
authors.lastname: Hoeben
authors.forename: Emmanuel Jhj
authors.initials: EJ
authors.lastname: Wiertz
mesh_terms: D000818:Animals; D015551:Autoimmunity; D002096:C-Peptide / Q000378:metabolism; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D003602:Cytotoxicity, Immunologic; D003922:Diabetes Mellitus, Type 1 / Q000276:immunology / Q000378:metabolism; D015870:Gene Expression; D023061:Gene Order; D005822:Genetic Vectors / Q000235:genetics; D015789:HLA-A2 Antigen / Q000276:immunology; D006801:Humans; D007328:Insulin / Q000235:genetics / Q000276:immunology; D050417:Insulin-Secreting Cells / Q000378:metabolism; D007515:Islets of Langerhans / Q000276:immunology* / Q000378:metabolism*; D016381:Islets of Langerhans Transplantation; D016086:Lentivirus / Q000235:genetics; D008297:Male; D051379:Mice; D009928:Organ Specificity / Q000235:genetics; D011401:Promoter Regions, Genetic; D011498:Protein Precursors / Q000276:immunology; D015843:Serpins / Q000235:genetics / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic; D014161:Transduction, Genetic; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology
chemical_list: D002096:C-Peptide; D015789:HLA-A2 Antigen; D007328:Insulin; D011498:Protein Precursors; D015843:Serpins; C118670:US2 protein, Varicellovirus; D014759:Viral Envelope Proteins; C013737:preproinsulin