title: Ex vivo priming of naïve T cells into EBV-specific Th1/Tc1 effector cells by mature autologous DC loaded with apoptotic/necrotic LCL.
abstract: Posttransplant lymphoproliferative disorders (PTLDs) represent life-threatening complications of bone marrow and solid organ transplantation (SOTx). These are B-cell malignancies triggered by Epstein-Barr Virus (EBV) infection in chronically immunosuppressed (IS) recipients. Immunosuppressed EBV seronegative (EBV(-)) organ recipients are at highest risk of developing PTLD owing to the lack of anti-EBV memory T cells to control subsequent EBV challenges. Our aim is to establish effective anti-EBV T-cell generation protocols for prevention or treatment of PTLD encountered in SOTx. We have used autologous dendritic cells (DCs) loaded with apoptotic/necrotic lymphoblastoid cell lines (LCLs) to evaluate the ability of such an approach to activate naïve T cells in vitro. In EBV(-) individuals, both CD8+ and CD4+ T-cell responses were amplified by this approach, as detected by IFN-gamma ELISPOT and cytotoxicity assays. The CD8+ T cells were poly-specific anti-EBNA3 A, -LMP2 and -BMLF1, with uniform reversion to a CD45RO+/RA-phenotype, decreased CD62L expression, and up-regulation of the activation markers CD28 and CD69. Addition of rhIL-12 improved anti-viral T-cell responses and reduced the functional differences observed between EBV(+) and EBV(-) responders. In conclusion, the DC/LCL method promotes cross-presentation of EBV-associated epitopes and may serve as an effective protocol for the adoptive immunotherapy of PTLD in EBV(-) SOTx patients.
authors.affiliation: Thomas E. Starzl Transplantation Institute, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: Iulia
authors.initials: I
authors.lastname: Popescu
authors.forename: Camila
authors.initials: C
authors.lastname: Macedo
authors.forename: Adriana
authors.initials: A
authors.lastname: Zeevi
authors.forename: Joseph
authors.initials: J
authors.lastname: Nellis
authors.forename: Kevin R
authors.initials: KR
authors.lastname: Patterson
authors.forename: Allison
authors.initials: A
authors.lastname: Logar
authors.forename: David
authors.initials: D
authors.lastname: Rowe
authors.forename: Jorge
authors.initials: J
authors.lastname: Reyes
authors.forename: Abdul S
authors.initials: AS
authors.lastname: Rao
authors.forename: Walter J
authors.initials: WJ
authors.lastname: Storkus
authors.forename: John J
authors.initials: JJ
authors.lastname: Fung
authors.forename: Diana
authors.initials: D
authors.lastname: Metes
mesh_terms: D017951:Antigen Presentation; D000941:Antigens / Q000378:metabolism; D015703:Antigens, CD / Q000096:biosynthesis; D000945:Antigens, Differentiation, T-Lymphocyte / Q000096:biosynthesis; D017209:Apoptosis; D018106:CD28 Antigens / Q000096:biosynthesis; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D003602:Cytotoxicity, Immunologic; D003713:Dendritic Cells / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D020031:Epstein-Barr Virus Infections / Q000276:immunology; D005434:Flow Cytometry; D006680:HLA Antigens / Q000096:biosynthesis; D004854:Herpesvirus 4, Human / Q000378:metabolism*; D006801:Humans; D007167:Immunotherapy / Q000379:methods; D016219:Immunotherapy, Adoptive; D019041:L-Selectin / Q000096:biosynthesis; D037181:Lectins, C-Type; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000378:metabolism; D008232:Lymphoproliferative Disorders / Q000209:etiology; D009000:Monocytes / Q000378:metabolism; D009336:Necrosis; D010455:Peptides / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000821:virology*; D018417:Th1 Cells / Q000821:virology*; D013997:Time Factors; D000099192:CD69 Antigens
chemical_list: D000941:Antigens; D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D018106:CD28 Antigens; D000939:Epitopes; D006680:HLA Antigens; D019041:L-Selectin; D037181:Lectins, C-Type; D010455:Peptides; D000099192:CD69 Antigens