A stromal cell free culture system generates mouse pro-T cells that can reconstitute T-cell compartments in vivo.

Publisher: Wiley-VCH Country of Publication: Germany NLM ID: 1273201 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1521-4141 (Electronic) Linking ISSN: 00142980 NLM ISO Abbreviation: Eur J Immunol Subsets: MEDLINE

Publication: <2005->: Weinheim : Wiley-VCH
Original Publication: Weinheim, Verlag Chemie GmbH.

Cell Culture Techniques/*methods ; Gene Rearrangement, alpha-Chain T-Cell Antigen Receptor/*immunology ; Gene Rearrangement, beta-Chain T-Cell Antigen Receptor/*immunology ; Precursor Cells, T-Lymphoid/*immunology ; Receptors, Antigen, T-Cell, alpha-beta/*immunology ; T-Lymphocytes, Regulatory/*immunology; Adaptor Proteins, Signal Transducing ; Animals ; Antigens, CD/genetics ; Antigens, CD/immunology ; CD3 Complex/genetics ; CD3 Complex/immunology ; Calcium-Binding Proteins ; Cells, Cultured ; Female ; Gene Rearrangement, alpha-Chain T-Cell Antigen Receptor/genetics ; Gene Rearrangement, beta-Chain T-Cell Antigen Receptor/genetics ; Intracellular Signaling Peptides and Proteins/genetics ; Intracellular Signaling Peptides and Proteins/immunology ; Membrane Proteins/genetics ; Membrane Proteins/immunology ; Mice ; Mice, Inbred BALB C ; Mice, Knockout ; Precursor Cells, T-Lymphoid/cytology ; Receptors, Antigen, T-Cell, alpha-beta/genetics ; Stromal Cells ; T-Lymphocytes, Regulatory/cytology

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.
(© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Keywords: BM transplantation; Lymphopenia; Notch ligand Delta-like 4 (DL4); T-cell development; Treg cell

0 (Adaptor Proteins, Signal Transducing)
0 (Antigens, CD)
0 (CD3 Complex)
0 (Calcium-Binding Proteins)
0 (Cd3e protein, mouse)
0 (DLL4 protein, mouse)
0 (Intracellular Signaling Peptides and Proteins)
0 (Membrane Proteins)
0 (Receptors, Antigen, T-Cell, alpha-beta)

Date Created: 20141120 Date Completed: 20150508 Latest Revision: 20201209

20231215

10.1002/eji.201444681

25408420