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Expansion of human regulatory T-cells from patients with type 1 diabetes.
- Source :
-
Diabetes [Diabetes] 2009 Mar; Vol. 58 (3), pp. 652-62. Date of Electronic Publication: 2008 Dec 15. - Publication Year :
- 2009
-
Abstract
- Objective: Regulatory T-cells (Tregs) have catalyzed the field of immune regulation. However, translating Treg-based therapies from animal models of autoimmunity to human clinical trials requires robust methods for the isolation and expansion of these cells-a need forming the basis for these studies.<br />Research Design and Methods: Tregs from recent-onset type 1 diabetic patients and healthy control subjects were isolated by fluorescence-activated cell sorting and compared for their capacity to expand in vitro in response to anti-CD3-anti-CD28-coated microbeads and IL-2. Expanded cells were examined for suppressive function, lineage markers and FOXP3, and cytokine production.<br />Results: Both CD4+CD127(lo/-) and CD4+CD127(lo/-)CD25+ T-cells could be expanded and used as Tregs. However, expansion of CD4+CD127(lo/-) cells required the addition of rapamycin to maintain lineage purity. In contrast, expansion of CD4+CD127(lo/-)CD25+ T-cells, especially the CD45RA+ subset, resulted in high yield, functional Tregs that maintained higher FOXP3 expression in the absence of rapamycin. Tregs from type 1 diabetic patients and control subjects expanded similarly and were equally capable of suppressing T-cell proliferation. Regulatory cytokines were produced by Tregs after culture; however, a portion of FOXP3+ cells were capable of producing interferon (IFN)-gamma after reactivation. IFN-gamma production was observed from both CD45RO+ and CD45RA+ Treg populations.<br />Conclusions: The results support the feasibility of isolating Tregs for in vitro expansion. Based on expansion capacity, FOXP3 stability, and functional properties, the CD4+CD127(lo/-)CD25+ T-cells represent a viable cell population for cellular therapy in this autoimmune disease.
- Subjects :
- Adult
Age of Onset
Antigens, CD immunology
CD4-Positive T-Lymphocytes immunology
Cell Division
Cell Proliferation
Cytokines metabolism
Female
Flow Cytometry
Forkhead Transcription Factors analysis
Humans
Immunosuppression Therapy
Ionomycin pharmacology
Male
Phenotype
Reference Values
T-Lymphocytes, Regulatory cytology
T-Lymphocytes, Regulatory drug effects
Tetradecanoylphorbol Acetate pharmacology
Young Adult
Diabetes Mellitus, Type 1 immunology
T-Lymphocytes, Regulatory immunology
Subjects
Details
- Language :
- English
- ISSN :
- 1939-327X
- Volume :
- 58
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- Diabetes
- Publication Type :
- Academic Journal
- Accession number :
- 19074986
- Full Text :
- https://doi.org/10.2337/db08-1168