title: A stromal cell free culture system generates mouse pro-T cells that can reconstitute T-cell compartments in vivo.
abstract: T-cell lymphopenia following BM transplantation or diseases such as AIDS result in immunodeficiency. Novel approaches to ameliorate this situation are urgently required. Herein, we describe a novel stromal cell free culture system in which Lineage(-) Sca1(+)c-kit(+) BM hematopoietic progenitors very efficiently differentiate into pro-T cells. This culture system consists of plate-bound Delta-like 4 Notch ligand and the cytokines SCF and IL-7. The pro-T cells developing in these cultures express CD25, CD117, and partially CD44; express cytoplasmic CD3ε; and have their TCRβ locus partially D-J rearranged. They could be expanded for over 3 months and used to reconstitute the T-cell compartments of sublethally irradiated T-cell-deficient CD3ε(-/-) mice or lethally irradiated WT mice. Pro-T cells generated in this system could partially correct the T-cell lymphopenia of pre-Tα(-/-) mice. However, reconstituted CD3ε(-/-) mice suffered from a wasting disease that was prevented by co-injection of purified CD4(+) CD25(high) WT Treg cells. In a T-cell-sufficient or T-lymphopenic setting, the development of disease was not observed. Thus, this in vitro culture system represents a powerful tool to generate large numbers of pro-T cells for transplantation and possibly with clinical applications.
authors.affiliation: Developmental and Molecular Immunology, Department of Biomedicine, University of Basel, Basel, Switzerland.
authors.forename: Nadine
authors.initials: N
authors.lastname: Gehre
authors.forename: Anja
authors.initials: A
authors.lastname: Nusser
authors.forename: Lilly
authors.initials: L
authors.lastname: von Muenchow
authors.forename: Roxane
authors.initials: R
authors.lastname: Tussiwand
authors.forename: Corinne
authors.initials: C
authors.lastname: Engdahl
authors.forename: Giuseppina
authors.initials: G
authors.lastname: Capoferri
authors.forename: Nabil
authors.initials: N
authors.lastname: Bosco
authors.forename: Rhodri
authors.initials: R
authors.lastname: Ceredig
authors.forename: Antonius G
authors.initials: AG
authors.lastname: Rolink
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D015703:Antigens, CD / Q000235:genetics / Q000276:immunology; D017252:CD3 Complex / Q000235:genetics / Q000276:immunology; D002135:Calcium-Binding Proteins; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D005260:Female; D015332:Gene Rearrangement, alpha-Chain T-Cell Antigen Receptor / Q000235:genetics / Q000276:immunology*; D015333:Gene Rearrangement, beta-Chain T-Cell Antigen Receptor / Q000235:genetics / Q000276:immunology*; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000276:immunology; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D018345:Mice, Knockout; D054504:Precursor Cells, T-Lymphoid / Q000166:cytology / Q000276:immunology*; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics / Q000276:immunology*; D017154:Stromal Cells; D050378:T-Lymphocytes, Regulatory / Q000166:cytology / Q000276:immunology*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D015703:Antigens, CD; D017252:CD3 Complex; D002135:Calcium-Binding Proteins; C485468:Cd3e protein, mouse; C519665:DLL4 protein, mouse; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D016693:Receptors, Antigen, T-Cell, alpha-beta
keywords: BM transplantation; Lymphopenia; Notch ligand Delta-like 4 (DL4); T-cell development; Treg cell