title: Exclusive Transduction of Human CD4+ T Cells upon Systemic Delivery of CD4-Targeted Lentiviral Vectors.
abstract: Playing a central role in both innate and adaptive immunity, CD4(+) T cells are a key target for genetic modifications in basic research and immunotherapy. In this article, we describe novel lentiviral vectors (CD4-LV) that have been rendered selective for human or simian CD4(+) cells by surface engineering. When applied to PBMCs, CD4-LV transduced CD4(+) but not CD4(-) cells. Notably, also unstimulated T cells were stably genetically modified. Upon systemic or intrasplenic administration into mice reconstituted with human PBMCs or hematopoietic stem cells, reporter gene expression was predominantly detected in lymphoid organs. Evaluation of GFP expression in organ-derived cells and blood by flow cytometry demonstrated exclusive gene transfer into CD4(+) human lymphocytes. In bone marrow and spleen, memory T cells were preferentially hit. Toward therapeutic applications, we also show that CD4-LV can be used for HIV gene therapy, as well as for tumor therapy, by delivering chimeric Ag receptors. The potential for in vivo delivery of the FOXP3 gene was also demonstrated, making CD4-LV a powerful tool for inducible regulatory T cell generation. In summary, our work demonstrates the exclusive gene transfer into a T cell subset upon systemic vector administration opening an avenue toward novel strategies in immunotherapy.
authors.affiliation: Molecular Biotechnology and Gene Therapy, Paul-Ehrlich-Institut, 63225 Langen, Germany;
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhou
authors.affiliation: Molecular Biotechnology and Gene Therapy, Paul-Ehrlich-Institut, 63225 Langen, Germany;
authors.forename: Katharina M
authors.initials: KM
authors.lastname: Uhlig
authors.affiliation: Molecular Biotechnology and Gene Therapy, Paul-Ehrlich-Institut, 63225 Langen, Germany;
authors.forename: Anke
authors.initials: A
authors.lastname: Muth
authors.affiliation: Division of Virology, Medical University of Innsbruck, A-6020 Innsbruck, Austria;
authors.forename: Janine
authors.initials: J
authors.lastname: Kimpel
authors.affiliation: Centre International de Recherche en Infectiologie, Virus Enveloppés, Vecteurs et Réponses Innées Équipe, INSERM U1111, Centre National de la Recherche Scientifique, Unités Mixtes de Recherche 5308, Université de Lyon-1, École Normale Supérieure de Lyon, 69007 Lyon, France;
authors.forename: Camille
authors.initials: C
authors.lastname: Lévy
authors.affiliation: Molecular Biotechnology and Gene Therapy, Paul-Ehrlich-Institut, 63225 Langen, Germany;
authors.forename: Robert C
authors.initials: RC
authors.lastname: Münch
authors.affiliation: Host Pathogen Interactions, Paul-Ehrlich-Institut, 63225 Langen, Germany;
authors.forename: Janna
authors.initials: J
authors.lastname: Seifried
authors.affiliation: Molecular Biotechnology and Gene Therapy, Paul-Ehrlich-Institut, 63225 Langen, Germany;
authors.forename: Anett
authors.initials: A
authors.lastname: Pfeiffer
authors.affiliation: Institute of Medical Virology, University of Zurich, CH-8057 Zurich, Switzerland;
authors.forename: Alexandra
authors.initials: A
authors.lastname: Trkola
authors.affiliation: Central Animal Unit, Paul-Ehrlich-Institut, 63225 Langen, Germany;
authors.forename: Cheick
authors.initials: C
authors.lastname: Coulibaly
authors.affiliation: Division of Virology, Medical University of Innsbruck, A-6020 Innsbruck, Austria;
authors.forename: Dorothee
authors.initials: D
authors.lastname: von Laer
authors.affiliation: Georg-Speyer-Haus, Institute for Tumor Biology and Experimental Therapy, D-60596 Frankfurt, Germany;
authors.forename: Winfried S
authors.initials: WS
authors.lastname: Wels
authors.affiliation: 3rd Department of Medicine-Hematology, Internal Oncology and Pneumology, University Medical Center of Johannes Gutenberg-University Mainz, 55131 Mainz, Germany;
authors.forename: Udo F
authors.initials: UF
authors.lastname: Hartwig
authors.affiliation: Centre International de Recherche en Infectiologie, Virus Enveloppés, Vecteurs et Réponses Innées Équipe, INSERM U1111, Centre National de la Recherche Scientifique, Unités Mixtes de Recherche 5308, Université de Lyon-1, École Normale Supérieure de Lyon, 69007 Lyon, France; INSERM U1065, Centre Méditerranéen de Médecine Moléculaire, Équipe 3, 06204 Nice, France; and.
authors.forename: Els
authors.initials: E
authors.lastname: Verhoeyen
authors.affiliation: Molecular Biotechnology and Gene Therapy, Paul-Ehrlich-Institut, 63225 Langen, Germany; German Cancer Consortium, 69120 Heidelberg, Germany christian.buchholz@pei.de.
authors.forename: Christian J
authors.initials: CJ
authors.lastname: Buchholz
mesh_terms: D000818:Animals; D001853:Bone Marrow / Q000378:metabolism; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism*; D045744:Cell Line, Tumor; D017690:Cell Transplantation / Q000379:methods; D002478:Cells, Cultured; D005434:Flow Cytometry; D051858:Forkhead Transcription Factors / Q000235:genetics / Q000378:metabolism; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods; D016086:Lentivirus / Q000235:genetics*; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D016688:Mice, Inbred NOD; D018345:Mice, Knockout; D016513:Mice, SCID; D013154:Spleen / Q000378:metabolism; D013950:Thymus Gland / Q000378:metabolism; D014161:Transduction, Genetic; D014183:Transplantation, Heterologous
chemical_list: C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; D049452:Green Fluorescent Proteins; D008156:Luciferases