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DnIKK2-Transfected Dendritic Cells Induce a Novel Population of Inducible Nitric Oxide Synthase???Expressing CD4+CD25??? Cells with Tolerogenic Properties
- Source :
- Transplantation. 83:474-484
- Publication Year :
- 2007
- Publisher :
- Ovid Technologies (Wolters Kluwer Health), 2007.
-
Abstract
- Background. We previously documented that rat bone marrow-derived dendritic cells (DCs), transfected with an adenovirus encoding a dominant negative form of IKK2 (dnIKK2), have impaired allostimulatory capacity and generate CD4 + T cells with regulatory function. Here we investigate the potency, the phenotype, and the mechanism of action of dnIKK2-DC-induced regulatory cells and we evaluated their tolerogenic properties in vivo. Methods. Brown Norway (BN) transfected dnIKK2-DCs were cultured with Lewis (LW) lymphocytes in primary mixed lymphocyte reaction (MLR). CD4 + T cells were purified from primary MLR and incubated in secondary coculture MLR with LW lymphocytes. Phenotypic characterization was performed by fluorescence-activated cell sorting and real-time polymerase chain reaction. The tolerogenic potential of CD4 + T cells pre-exposed to dnIKK2-DCs was evaluated in vivo in a model of kidney allotransplantation. Results. CD4 + T cells pre-exposed to dnIKK2-DCs were cD4 + CD25 -/dim and expressed interleukin (IL)-10, transforming growth factor-beta, interferon-gamma, IL-2, and inducible nitric oxide synthase (iNOS). These cells (dnIKK2-Treg), cocultured (at up to 1:10 5 ratio) with a primary MLR, suppressed T-cell proliferation to alloantigens. The regulatory effect was cell-to-cell contact-independent since it was also observed in a transwell system. A nitric oxide synthase inhibitor significantly reverted dnIKK2-Treg-mediated suppression, whereas neutralizing antibodies to IL-10 and TGF-beta had no significant effect. DnIKK2-Treg given in vivo to LW rats prolonged the survival of a kidney allograft from BN rats (the donor rat strain used for generating DCs). Conclusions. DnIKK2-Treg is a unique population of CD4 + CD25 - T cells expressing high levels of iNOS. These cells potently inhibit T-cell response in vitro and induce prolongation of kidney allograft survival in vivo.
- Subjects :
- CD4-Positive T-Lymphocytes
medicine.medical_specialty
Population
Nitric Oxide Synthase Type II
chemical and pharmacologic phenomena
Immune tolerance
In vivo
Internal medicine
Immune Tolerance
medicine
Animals
Transplantation, Homologous
IL-2 receptor
Antigen-presenting cell
education
Cells, Cultured
Transplantation
education.field_of_study
biology
Graft Survival
Interleukin-2 Receptor alpha Subunit
Cell Differentiation
hemic and immune systems
Dendritic Cells
Transfection
Dendritic cell
Kidney Transplantation
Molecular biology
Coculture Techniques
I-kappa B Kinase
Rats
Nitric oxide synthase
Phenotype
Endocrinology
Solubility
biology.protein
Subjects
Details
- ISSN :
- 00411337
- Volume :
- 83
- Database :
- OpenAIRE
- Journal :
- Transplantation
- Accession number :
- edsair.doi.dedup.....8831206997b5fffce25c4f3843c48d7c