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.
Original language | English |
---|---|
Pages (from-to) | 1592-1601 |
Number of pages | 10 |
Journal | Molecular therapy : the journal of the American Society of Gene Therapy |
Volume | 21 |
Issue number | 8 |
DOIs | |
Publication status | Published - Aug 2013 |
Keywords
- Animals
- Autoimmunity
- C-Peptide
- CD8-Positive T-Lymphocytes
- Cytotoxicity, Immunologic
- Diabetes Mellitus, Type 1
- Gene Expression
- Gene Order
- Genetic Vectors
- HLA-A2 Antigen
- Humans
- Insulin
- Insulin-Secreting Cells
- Islets of Langerhans
- Islets of Langerhans Transplantation
- Lentivirus
- Male
- Mice
- Organ Specificity
- Promoter Regions, Genetic
- Protein Precursors
- Serpins
- T-Lymphocytes, Cytotoxic
- Transduction, Genetic
- Viral Envelope Proteins