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OP9-DL1

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https://www.readbyqxmd.com/read/27852740/temporal-expression-of-bim-limits-the-development-of-agonist-selected-thymocytes-and-skews-their-tcr%C3%AE-repertoire
#1
Kun-Po Li, Anke Fähnrich, Eron Roy, Carla M Cuda, H Leighton Grimes, Harris R Perlman, Kathrin Kalies, David A Hildeman
CD8αα TCRαβ(+) intestinal intraepithelial lymphocytes play a critical role in promoting intestinal homeostasis, although mechanisms controlling their development and peripheral homeostasis remain unclear. In this study, we examined the spatiotemporal role of Bim in the thymic selection of CD8αα precursors and the fate of these cells in the periphery. We found that T cell-specific expression of Bim during early/cortical, but not late/medullary, thymic development controls the agonist selection of CD8αα precursors and limits their private TCRβ repertoire...
November 16, 2016: Journal of Immunology: Official Journal of the American Association of Immunologists
https://www.readbyqxmd.com/read/27787805/developmental-and-functional-assays-to-study-murine-and-human-%C3%AE-%C3%AE-t-cells
#2
Julie C Ribot, Karine Serre, Bruno Silva-Santos
The key roles played by gamma-delta (γδ) T cells in immunity to infection and tumors critically depend on their differentiation into effectors capable of secreting cytokines (such as interferon-γ or interleukin-17), and killing infected or transformed cells. Here we detail the main methods used to investigate the differentiation of γδ T cells from murine or human origin. We describe developmental assays, such as thymic organ cultures (TOCs) and coculture of progenitors cells with OP9-DL1 stomal cells, as well as functional assays typically employed to evaluate γδ T cell cytotoxicity and cytokine production...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27251160/tnf-alpha-and-notch-signaling-regulates-the-expression-of-hoxb4-and-gata3-during-early-t-lymphopoiesis
#3
Josiane Lilian Dos Santos Schiavinato, Lucila Habib Bourguignon Oliveira, Amélia Goes Araujo, Maristela Delgado Orellana, Patrícia Viana Bonini de Palma, Dimas Tadeu Covas, Marco Antonio Zago, Rodrigo Alexandre Panepucci
During the early thymus colonization, Notch signaling activation on hematopoietic progenitor cells (HPCs) drives proliferation and T cell commitment. Although these processes are driven by transcription factors such as HOXB4 and GATA3, there is no evidence that Notch directly regulates their transcription. To evaluate the role of NOTCH and TNF signaling in this process, human CD34(+) HPCs were cocultured with OP9-DL1 cells, in the presence or absence of TNF. The use of a Notch signaling inhibitor and a protein synthesis inhibitor allowed us to distinguish primary effects, mediated by direct signaling downstream Notch and TNF, from secondary effects, mediated by de novo synthesized proteins...
October 2016: In Vitro Cellular & Developmental Biology. Animal
https://www.readbyqxmd.com/read/26777320/melanoma-immunotherapy-in-mice-using-genetically-engineered-pluripotent-stem-cells
#4
Mohammad Haque, Jianyong Song, Kristin Fino, Praneet Sandhu, Youfei Wang, Bing Ni, Deyu Fang, Jianxun Song
Adoptive cell transfer (ACT) of antigen (Ag)-specific CD8(+) cytotoxic T lymphocytes (CTLs) is a highly promising treatment for a variety of diseases. Naive or central memory T-cell-derived effector CTLs are optimal populations for ACT-based immunotherapy because these cells have a high proliferative potential, are less prone to apoptosis than terminally differentiated cells, and have the higher ability to respond to homeostatic cytokines. However, such ACT with T-cell persistence is often not feasible due to difficulties in obtaining sufficient cells from patients...
2016: Cell Transplantation
https://www.readbyqxmd.com/read/25569376/invariant-natural-killer-t-cells-generated-from-human-adult-hematopoietic-stem-progenitor-cells-are-poly-functional
#5
Wenji Sun, Yi Wang, James E East, Amy S Kimball, Katherine Tkaczuk, Susan Kesmodel, Scott E Strome, Tonya J Webb
Invariant natural killer T (iNKT) cells constitute an important subset of T cells that can both directly and indirectly mediate anti-tumor immunity. However, cancer patients have a reduction in both iNKT cell number and function, and these deficits limit the potential clinical application of iNKT cells for cancer therapy. To overcome the problem of limited iNKT cell numbers, we investigated whether iNKT cells can be generated in vitro from bone marrow-derived adult hematopoietic stem-progenitor cells (HSPC)...
March 2015: Cytokine
https://www.readbyqxmd.com/read/25349888/derivation-of-t-cells-in-vitro-from-mouse-embryonic-stem-cells
#6
Martina Kučerová-Levisohn, Jordana Lovett, Armin Lahiji, Roxanne Holmes, Juan Carlos Zúñiga-Pflücker, Benjamin D Ortiz
The OP9/OP9-DL1 co-culture system has become a well-established method for deriving differentiated blood cell types from embryonic and hematopoietic progenitors of both mouse and human origin. It is now used to address a growing variety of complex genetic, cellular and molecular questions related to hematopoiesis, and is at the cutting edge of efforts to translate these basic findings to therapeutic applications. The procedures are straightforward and routinely yield robust results. However, achieving successful hematopoietic differentiation in vitro requires special attention to the details of reagent and cell culture maintenance...
2014: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/25329250/small-molecule-inhibitor-of-glycogen-synthase-kinase-3%C3%AE-6-bromoindirubin-3-oxime-inhibits-hematopoietic-regeneration-in-stem-cell-recipient-mice
#7
Sylvie Shen, Ning Xu, Guy Klamer, Kap-Hyoun Ko, Melissa Khoo, David Ma, John Moore, Tracey A O'Brien, Alla Dolnikov
Small-molecule inhibitors of glycogen synthase kinase 3β (GSK3β) have demonstrated strong anti-leukemia effects in preclinical studies. Here, we investigated the effect of GSK3β inhibitor 6-Bromoindirubin-3-oxime (BIO) previously shown to inhibit leukemia cell growth in vitro and of animal models on hematopoietic regeneration in recipients of stem cell transplant. BIO administered to immunocompromised mice transplanted with human hematopoietic stem cells inhibited human stem cell engraftment in the bone marrow (BM) and peripheral blood...
March 15, 2015: Stem Cells and Development
https://www.readbyqxmd.com/read/25309961/the-dn2-myeloid-t-dn2mt-progenitor-is-a-target-cell-for-leukemic-transformation-by-the-tlx1-oncogene
#8
Lynnsey A Zweier-Renn, Irene Riz, Teresa S Hawley, Robert G Hawley
INTRODUCTION: Inappropriate activation of the TLX1 (T-cell leukemia homeobox 1) gene by chromosomal translocation is a recurrent event in human T-cell Acute Lymphoblastic Leukemia (T-ALL). Ectopic expression of TLX1 in murine bone marrow progenitor cells using a conventional retroviral vector efficiently yields immortalized cell lines and induces T-ALL-like tumors in mice after long latency. METHODS: To eliminate a potential contribution of retroviral insertional mutagenesis to TLX1 immortalizing and transforming function, we incorporated the TLX1 gene into an insulated self-inactivating retroviral vector...
February 20, 2013: Journal of Bone Marrow Research
https://www.readbyqxmd.com/read/24889652/gene-profiling-reveals-association-between-altered-wnt-signaling-and-loss-of-t-cell-potential-with-age-in-human-hematopoietic-stem-cells
#9
Melissa L M Khoo, Stephen M Carlin, Mark A Lutherborrow, Vivek Jayaswal, David D F Ma, John J Moore
Functional decline of the hematopoietic system occurs during aging and contributes to clinical consequences, including reduced competence of adaptive immunity and increased incidence of myeloid diseases. This has been linked to aging of the hematopoietic stem cell (HSC) compartment and has implications for clinical hematopoietic cell transplantation as prolonged periods of T-cell deficiency follow transplantation of adult mobilized peripheral blood (PB), the primary transplant source. Here, we examined the gene expression profiles of young and aged HSCs from human cord blood and adult mobilized PB, respectively, and found that Wnt signaling genes are differentially expressed between young and aged human HSCs, with less activation of Wnt signaling in aged HSCs...
August 2014: Aging Cell
https://www.readbyqxmd.com/read/24620791/infected-hematopoietic-stem-cells-and-with-integrated-hbv-dna-generate-defective-t-cells-in-chronic-hbv-infection-patients
#10
Y Shi, Y Lan, F Cao, Y Teng, L Li, F Wang, J Li, J Zhou, Y Li
A weak T-cell response plays a key role in the persistence of hepatitis B virus (HBV) infection. We aimed to confirm that T-cell defects in patients with chronic HBV infection are associated with HBV DNA infection of bone marrow (BM) hematopoietic stem cells (HSCs). Using reverse transcription polymerase chain reaction (RT-PCR) and fluorescence in situ hybridization (FISH), we observed the transcription of HBsAg coding genes and confirmed the integration of HBV DNA in CD34(+) BM HSCs from chronic HBV infection patients...
July 2014: Journal of Viral Hepatitis
https://www.readbyqxmd.com/read/24238609/generation-of-hematopoietic-stem-cells-from-purified-embryonic-endothelial-cells-by-a-simple-and-efficient-strategy
#11
Zhuan Li, Fan Zhou, Dongbo Chen, Wenyan He, Yanli Ni, Lingfei Luo, Bing Liu
Recent progress by versatile approaches supports the new hypothesis that multi-potent hematopoietic stem cells (HSCs) are directly formed from a rare population of endothelial cells in mid-gestation mouse embryos. This process is therefore known as the endothelial-to-hematopoietic transition (EHT). Nevertheless, there is no functional evidence that documents the HSC transition from purified endothelial cells. In this study, we developed an OP9-DL1-based co-culture system that was able to facilitate the HSC specification and/or expansion in vitro of mouse embryonic day 10...
November 20, 2013: Journal of Genetics and Genomics, Yi Chuan Xue Bao
https://www.readbyqxmd.com/read/24096122/the-orphan-nuclear-receptor-ear-2-nr2f6-is-a-novel-negative-regulator-of-t-cell-development
#12
Christine V Ichim, Džana D Dervović, Juan Carlos Zúñiga-Pflücker, Richard A Wells
We describe a novel role for the orphan nuclear receptor Ear-2 in regulating T cell development. Retrovirus-mediated overexpression of Ear-2 (EAR-2++) in a bone marrow (BM) transplantation assay resulted in limited T cell development and a greater than tenfold decrease in thymus size and cellularity relative to controls. Ear-2-transduced murine BM hematopoietic stem cells (HSCs) in OP9-DL1 cultures showed a proliferation deficit during days 1-5 after induction of differentiation, which corresponded to increased expression of the cell cycle regulators p21 (cdkn1a) and p27 (cdkn1b), as well as increased expression of Hes1, Notch3, Egr1, and Scl (Tal1) and decreased expression of Gli1, Gfi-1, HoxA9, PU...
January 2014: Experimental Hematology
https://www.readbyqxmd.com/read/24091848/in-vitro-generation-of-mature-naive-antigen-specific-cd8-t-cells-with-a-single-t-cell-receptor-by-agonist-selection
#13
S Snauwaert, G Verstichel, S Bonte, G Goetgeluk, S Vanhee, Y Van Caeneghem, K De Mulder, C Heirman, H Stauss, M H M Heemskerk, T Taghon, G Leclercq, J Plum, A W Langerak, K Thielemans, T Kerre, B Vandekerckhove
Peripheral blood T cells transduced with a tumor-specific T-cell receptor (TCR) face problems of auto-reactivity and lack of efficacy caused by cross-pairing of exogenous and endogenous TCR chains, as well as short term in vivo survival due to activation and growth factor-induced differentiation. We here studied an alternative strategy for the efficient generation of naive CD8(+) T cells with a single TCR. TCR-transduced human postnatal thymus-derived and adult mobilized blood-derived hematopoietic progenitor cells (HPCs) were differentiated to CD4(+)CD8(+) double-positive T cells using OP9-Delta-like 1 (OP9-DL1) cultures...
April 2014: Leukemia: Official Journal of the Leukemia Society of America, Leukemia Research Fund, U.K
https://www.readbyqxmd.com/read/23988616/induction-of-t-cell-development-by-delta-like-4-expressing-fibroblasts
#14
Mahmood Mohtashami, Divya K Shah, Korosh Kianizad, Geneve Awong, Juan Carlos Zúñiga-Pflücker
The thymus provides a unique environment for the induction of T-cell lineage commitment and differentiation, which is predicted by specific Notch ligand-receptor interactions on epithelial cells and lymphoid progenitors, respectively. Accordingly, a bone marrow-derived stromal cell line (OP9) ectopically expressing the Notch ligand Delta-like 1 (Dll1) or Dll4 (OP9-DL1 and OP9-DL4, respectively) gains the ability to recapitulate thymus-like function, supporting T-cell differentiation of both mouse and human progenitors...
October 2013: International Immunology
https://www.readbyqxmd.com/read/23988615/removal-of-myeloid-cytokines-from-the-cellular-environment-enhances-t-cell-development-in-vitro
#15
Monique F M A Smeets, Charley Mackenzie-Kludas, Mahmood Mohtashami, Hui-Hua Zhang, Juan Carlos Zúñiga-Pflücker, David J Izon
The majority of T-cell development occurs in the thymus. Thymic epithelial cells are specialized cells that express NOTCH ligands and secrete specific cytokines required for normal T-cell lymphopoiesis. It has been demonstrated that OP9 cells derived from macrophage colony-stimulating factor (M-CSF)-deficient mice can support T-cell development when transduced with a NOTCH ligand, Delta-like 1 (Dll1). In this report, we have tested CSF-deficient mouse fibroblasts transduced with Dll1 for their ability to support T-cell differentiation...
October 2013: International Immunology
https://www.readbyqxmd.com/read/23851691/cellular-and-molecular-requirements-for-the-selection-of-in-vitro-generated-cd8-t-cells-reveal-a-role-for-notch
#16
Dzana D Dervovic, Haydn C-Y Liang, Jennifer L Cannons, Alisha R Elford, Mahmood Mohtashami, Pamela S Ohashi, Pamela L Schwartzberg, Juan Carlos Zúñiga-Pflücker
Differentiation of CD8 single-positive (SP) T cells is predicated by the ability of lymphocyte progenitors to integrate multiple signaling cues provided by the thymic microenvironment. In the thymus and the OP9-DL1 system for T cell development, Notch signals are required for progenitors to commit to the T cell lineage and necessary for their progression to the CD4(+)CD8(+) double-positive (DP) stage of T cell development. However, it remains unclear whether Notch is a prerequisite for the differentiation of DP cells to the CD8 SP stage of development...
August 15, 2013: Journal of Immunology: Official Journal of the American Association of Immunologists
https://www.readbyqxmd.com/read/23756946/directed-differentiation-of-embryonic-stem-cells-to-the-t-lymphocyte-lineage
#17
Haydn C-Y Liang, Roxanne Holmes, Juan Carlos Zúñiga-Pflücker
Hematopoiesis is the highly regulated and complex process by which blood cells are formed. Hematopoiesis can be achieved in vitro by the differentiation of embryonic stem cells (ESCs) into hematopoietic lineage cells. Differentiation of ESCs initially gives rise to mesoderm colonies that go on to form hemangioblast cells, which possess endothelial and hematopoietic lineage potential. While the differentiation of several hematopoietic lineages from ESCs, such as erythrocytes and macrophages, can be easily recapitulated in vitro, T-cell differentiation requires additional Notch-dependent signals for their generation...
2013: Methods in Molecular Biology
https://www.readbyqxmd.com/read/23670190/age-associated-changes-in-the-differentiation-potentials-of-human-circulating-hematopoietic-progenitors-to-t-or-nk-lineage-cells
#18
Seishi Kyoizumi, Yoshiko Kubo, Junko Kajimura, Kengo Yoshida, Kazue Imai, Tomonori Hayashi, Kei Nakachi, Lauren F Young, Malcolm A Moore, Marcel R M van den Brink, Yoichiro Kusunoki
Age-associated changes of T and NK cell (T/NK) potential of human hematopoietic stem cells are unknown. In this study, we enumerate and characterize T/NK precursors among CD34(+)Lin(-) cell populations circulating in normal human adult peripheral blood (PB) by a limiting-dilution assay using coculture with OP9-DL1 stroma cells expressing Notch 1 ligand, Delta-like 1. The frequency of T cell precursors in CD34(+)Lin(-) cells was found to decrease with donor age, whereas the ratio of NK to T cell precursor frequency (NK/T ratio) increased with age, suggesting that lymphoid differentiation potential of PB progenitors shifts from T to NK cell lineage with aging...
June 15, 2013: Journal of Immunology: Official Journal of the American Association of Immunologists
https://www.readbyqxmd.com/read/23587923/cd34-expansion-with-delta-1-and-hoxb4-promotes-rapid-engraftment-and-transfusion-independence-in-a-macaca-nemestrina-cord-blood-transplant-model
#19
Korashon L Watts, Colleen Delaney, Veronica Nelson, Grant D Trobridge, Brian C Beard, R Keith Humphries, Hans-Peter Kiem
Umbilical cord blood (CB) transplantation is a promising therapeutic approach but continues to be associated with delayed engraftment and infections. Here, we explored in our macaque CB transplant model expansion and engraftment kinetics of cells expanded with the combination of HOXB4 and Delta-1. CB cells were divided into two equal fractions; one fraction was transduced with HOXB4 yellow fluorescent protein (YFP) and expanded on control OP9 cells, and the other was transduced with HOXB4 green fluorescent protein (GFP) and expanded on Delta-expressing OP9 cells (OP9-DL1)...
June 2013: Molecular Therapy: the Journal of the American Society of Gene Therapy
https://www.readbyqxmd.com/read/23378057/lmo2-induces-hematopoietic-stem-cell-like-features-in-t-cell-progenitor-cells-prior-to-leukemia
#20
Susan M Cleveland, Stephen Smith, Rati Tripathi, Elizabeth M Mathias, Charnise Goodings, Natalina Elliott, Dunfa Peng, Wael El-Rifai, Dajun Yi, Xi Chen, Liqi Li, Charles Mullighan, James R Downing, Paul Love, Utpal P Davé
LIM domain only 2 (Lmo2) is frequently deregulated in sporadic and gene therapy-induced acute T-cell lymphoblastic leukemia (T-ALL) where its overexpression is an important initiating mutational event. In transgenic and retroviral mouse models, Lmo2 expression can be enforced in multiple hematopoietic lineages but leukemia only arises from T cells. These data suggest that Lmo2 confers clonal growth advantage in T-cell progenitors. We analyzed proliferation, differentiation, and cell death in CD2-Lmo2 transgenic thymic progenitor cells to understand the cellular effects of enforced Lmo2 expression...
May 2013: Stem Cells
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