title: Manipulation of human early T lymphopoiesis by coculture on human bone marrow stromal cells: potential utility for adoptive immunotherapy.
abstract: T cell precursors are an attractive target for adoptive immunotherapy. We examined the regulation of human early T lymphopoiesis by human bone marrow stromal cells to explore in vitro manipulation of human T cell precursors in a human-only coculture system. The generation of CD7(+)CD56(-)cyCD3(-) proT cells from human hematopoietic progenitors on telomerized human bone marrow stromal cells was enhanced by stem cell factor, flt3 ligand, and thrombopoietin, but these stimulatory effects were suppressed by interleukin 3. Expression of Notch ligands Delta-1 and -4 on stromal cells additively promoted T cell differentiation into the CD7(+)cyCD3(+) pre-T cell stage, while cell growth was strongly inhibited. By combining these coculture systems, we found that initial coculture with telomerized stromal cells in the presence of stem cell factor, flt3 ligand, and thrombopoietin, followed by coculture on Delta-1- and -4-coexpressing stromal cells led to a higher percentage and number of pre-T cells. Adoptive immunotherapy using peripheral blood T cells transduced with a tumor antigen-specific T cell receptor (TCR) is a promising strategy but has several limitations, such as the risk of forming a chimeric TCR with the endogenous TCR. We demonstrated that incubation of TCR-transduced hematopoietic progenitors with the combination of coculture systems gave rise to CD7(+)TCR(+)CD3(+)CD1a(-) T cell precursors that rapidly proliferated and differentiated under the culture condition to induce mature T cell differentiation. These data show the regulatory mechanism of early T lymphopoiesis on human stromal cells and the potential utility of engineered human stromal cells to manipulate early T cell development for clinical application.
authors.affiliation: Hematology and Oncology, Mie University Graduate School of Medicine, Tsu, Mie, Japan.
authors.forename: Bing
authors.initials: B
authors.lastname: Liu
authors.forename: Kohshi
authors.initials: K
authors.lastname: Ohishi
authors.forename: Yuki
authors.initials: Y
authors.lastname: Orito
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Nakamori
authors.forename: Hiroyoshi
authors.initials: H
authors.lastname: Nishikawa
authors.forename: Kazuko
authors.initials: K
authors.lastname: Ino
authors.forename: Kei
authors.initials: K
authors.lastname: Suzuki
authors.forename: Takeshi
authors.initials: T
authors.lastname: Matsumoto
authors.forename: Masahiro
authors.initials: M
authors.lastname: Masuya
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Hamada
authors.forename: Junichi
authors.initials: J
authors.lastname: Mineno
authors.forename: Ryoichi
authors.initials: R
authors.lastname: Ono
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Nosaka
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Shiku
authors.forename: Naoyuki
authors.initials: N
authors.lastname: Katayama
mesh_terms: D000818:Animals; D019011:Antigens, CD7 / Q000378:metabolism; D015153:Blotting, Western; D017252:CD3 Complex / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D002460:Cell Line; D002478:Cells, Cultured; D018920:Coculture Techniques; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods; D007377:Interleukin-3 / Q000494:pharmacology; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D038041:Lymphopoiesis; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000378:metabolism; D019089:Stem Cell Factor / Q000494:pharmacology; D013601:T-Lymphocytes / Q000166:cytology* / Q000378:metabolism; D016615:Telomere / Q000235:genetics / Q000378:metabolism; D013926:Thrombopoietin / Q000494:pharmacology; D013997:Time Factors
chemical_list: D019011:Antigens, CD7; D017252:CD3 Complex; D007377:Interleukin-3; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D011948:Receptors, Antigen, T-Cell; D019089:Stem Cell Factor; C064045:delta protein; C084881:flt3 ligand protein; D049452:Green Fluorescent Proteins; D013926:Thrombopoietin