title: Preconditioning therapy with lentiviral vector-programmed dendritic cells accelerates the homeostatic expansion of antigen-reactive human T cells in NOD.Rag1-/-.IL-2rγc-/- mice.
abstract: Dendritic cell (DC)-based immunization is a potent strategy to direct prompt and durable immune responses against viral reactivations after transplantations. Here, we show that overnight lentiviral vector (LV) gene transfer into human monocytes co-expressing granulocyte-macrophage colony stimulating factor and interleukin (IL)-4 induced self-differentiated DCs (SMART-DCs) with stable DC immunophenotype over weeks in culture and secreted several inflammatory cytokines. SMART-DCs injected subcutaneously in immunodeficient NOD.Rag1(-/-).IL2rγ(-/-) (NRG) mice 1 day after LV transduction were stable for a month in vivo. "Conventional" DCs (cDCs) and SMART-DCs were compared with regard to their potency to accelerate the expansion, biodistribution, and antigenic stimulation of autologous human T cells. Peripheral blood cells obtained from human cytomegalovirus (hCMV)-reactive donors and full-length hCMV pp65 antigenic protein or peptides were used. DCs loaded with pp65 were administered subcutaneously into NRG mice as a preconditioning treatment a week prior to intravenous infusion with T cells. Optical imaging analyses demonstrated that in mice preconditioned with SMART-DC-pp65, T cells were directly recruited to the immunization site and subsequently spread to the spleen and other organs. A dramatic expansion of both human CD8(+) and CD4(+) T cells could be observed within a few days after infusion, and this was associated with consistent measurable CD8(+) effector memory T-cell responses against different pp65 epitopes. Thus, this mouse model demonstrates the proof-of-principle for SMART-DCs to accelerate expansion of human lymphocytes, resulting in poly-functional and antigen-specific immune responses against hCMV-pp65.
authors.affiliation: Department of Hematology, Hemostasis, Oncology and Stem Cell Transplantation, Hannover Medical School, Carl Neuberg Strasse 1, Hannover, Germany .
authors.forename: Gustavo
authors.initials: G
authors.lastname: Salguero
authors.forename: Bala Sai
authors.initials: BS
authors.lastname: Sundarasetty
authors.forename: Sylvia
authors.initials: S
authors.lastname: Borchers
authors.forename: Dirk
authors.initials: D
authors.lastname: Wedekind
authors.forename: Britta
authors.initials: B
authors.lastname: Eiz-Vesper
authors.forename: Sarvari
authors.initials: S
authors.lastname: Velaga
authors.forename: Adan C
authors.initials: AC
authors.lastname: Jirmo
authors.forename: Georg
authors.initials: G
authors.lastname: Behrens
authors.forename: Gregor
authors.initials: G
authors.lastname: Warnecke
authors.forename: Ann-Kathrin
authors.initials: AK
authors.lastname: Knöfel
authors.forename: Rainer
authors.initials: R
authors.lastname: Blasczyk
authors.forename: Eva
authors.initials: E
authors.lastname: Mischak-Weissinger
authors.forename: Arnold
authors.initials: A
authors.lastname: Ganser
authors.forename: Renata
authors.initials: R
authors.lastname: Stripecke
mesh_terms: D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002454:Cell Differentiation / Q000276:immunology; D049109:Cell Proliferation; D003713:Dendritic Cells / Q000276:immunology* / Q000378:metabolism / Q000637:transplantation*; D016654:Genes, RAG-1 / Q000235:genetics; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007156:Immunologic Memory / Q000276:immunology; D007167:Immunotherapy / Q000379:methods*; D053631:Interleukin Receptor Common gamma Subunit / Q000235:genetics; D016086:Lentivirus; D008231:Lymphopenia / Q000628:therapy*; D051379:Mice; D016688:Mice, Inbred NOD; D018345:Mice, Knockout; D009000:Monocytes / Q000166:cytology; D009894:Opportunistic Infections / Q000517:prevention & control* / Q000821:virology; D010750:Phosphoproteins / Q000276:immunology; D014161:Transduction, Genetic; D014763:Viral Matrix Proteins / Q000276:immunology
chemical_list: D053631:Interleukin Receptor Common gamma Subunit; D010750:Phosphoproteins; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa