Dendritic cells genetically engineered with adenoviral vector encoding dnIKK2 induce the formation of potent CD4+ t-regulatory cells

Susanna Tomasoni, Sistiana Aiello, Linda Cassis, Marina Noris, Lorena Longaretti, Regiane A. Cavinato, Nadia Azzollini, Anna Pezzotta, Giuseppe Remuzzi, Ariela Benigni

Research output: Contribution to journalArticle

Abstract

Background. Immature dendritic cells (DC), characterized by low expression of both major histocompatibility complex class II antigens and co-stimulatory molecules, can be instrumental in the induction of peripheral tolerance. Because nuclear factor (NF)-κB is central to DC maturation, the authors engineered DC with an adenoviral vector (Adv) encoding for a kinase-defective dominant negative form of IKK2 (dnIKK2) to block NF-κB activation and inhibit DC maturation. Methods. DC were obtained by culturing bone marrow from Brown Norway (BN) rats with granulocyte-macrophage colony-stimulating factor and interleukin-4 for 11 days. To block NF-κB activation, at day 9, cells were transfected with AdV-dnIKK2. At day 11, cells were used as stimulators in primary mixed leukocyte reaction (MLR) with naive Lewis rat lymphocytes as responders. CD4+ T cells were purified from primary MLR and tested in secondary MLR with allogeneic mature DC and in co-culture MLR with naive lymphocytes. The tolerogenic potential of dnIKK2-DC was evaluated in vivo in a model of rat kidney allotransplantation. Results. DnIKK2-DC were immature and lacked any allostimulatory activity. T cells preexposed to allogeneic dnIKK2-DC were hyporesponsive to a secondary stimulation with mature DC and acquired potent regulatory properties, inhibiting naive T-cell proliferation toward allogeneic stimuli. Pretransplant infusion of allogeneic donor dnIKK2-DC prolonged the survival of a kidney allograft from the same allogeneic donor, without the need for immunosuppressive therapy. Conclusions. Allogeneic DC, rendered immature by dnIKK2 transfection, induce in vitro differentiation of naive T cells into CD4+ T-regulatory cells, effective at low ratios with target cells, rendering them applicable for cellular therapy of immune-mediated abnormalities and for preventing transplant rejection.

Original languageEnglish
Pages (from-to)1056-1061
Number of pages6
JournalTransplantation
Volume79
Issue number9
DOIs
Publication statusPublished - May 15 2005

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Keywords

  • Dendritic cells
  • Nuclear factor-κB
  • T-regulatory cells

ASJC Scopus subject areas

  • Transplantation
  • Immunology

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