title: In vivo and ex vivo gene transfer in thymocytes and thymocyte precursors.
abstract: The thymus provides a specialized environment allowing the differentiation of T lymphocytes from bone marrow-derived progenitor cells. We and others have demonstrated that gene transfer into distinct thymocyte populations can be obtained, both in vivo and ex vivo, using lentiviral vectors. Here, we describe techniques for intrathymic lentiviral transduction in mice, using a surgical approach wherein the thoracic cavity is exposed as well as a significantly less invasive strategy wherein virions are directly injected through the skin. Moreover, thymocyte differentiation from murine and human progenitors is now feasible in vitro, under conditions wherein the Notch and IL-7 signaling pathways are activated. We describe methods allowing transduction of murine and human progenitors and their subsequent differentiation into more mature thymocytes. Conditions for lentiviral gene transfer into more differentiated human thymocyte subsets are also presented. Optimization of technologies for HIV-based gene transfer into murine and human thymocyte progenitors will advance strategies aimed at modulating T-cell differentiation and function in-vivo; approaches potentially targeting patients with genetic and acquired immunodeficiencies as well as immune-sensitive tumors. Furthermore, this technology will foster the progression of basic research aimed at elucidating molecular aspects of T-cell differentiation in mice and humans.
authors.affiliation: Institut de Génétique Moléculaire de Montpellier, CNRS UMR 5535, Montpellier, IFR122, France.
authors.forename: Oumeya
authors.initials: O
authors.lastname: Adjali
authors.forename: Amélie
authors.initials: A
authors.lastname: Montel-Hagen
authors.forename: Louise
authors.initials: L
authors.lastname: Swainson
authors.forename: Sophie
authors.initials: S
authors.lastname: Marty
authors.forename: Rita
authors.initials: R
authors.lastname: Vicente
authors.forename: Cedric
authors.initials: C
authors.lastname: Mongellaz
authors.forename: Chantal
authors.initials: C
authors.lastname: Jacquet
authors.forename: Valérie
authors.initials: V
authors.lastname: Zimmermann
authors.forename: Naomi
authors.initials: N
authors.lastname: Taylor
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000276:immunology; D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers; D018014:Gene Transfer Techniques; D006801:Humans; D051379:Mice; D013950:Thymus Gland / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D014161:Transduction, Genetic
chemical_list: D018952:Antigens, CD34; D017931:DNA Primers