scholarly journals An in vitro system of T cell maturation-effect of cytokines and of thymic epithelial cells ♦ 865

1998 ◽  
Vol 43 ◽  
pp. 150-150
Author(s):  
Athena P Kourtis ◽  
Francis K Lee ◽  
Christian Ibegbu ◽  
Edmund Waller ◽  
Andre J Nahmias
Keyword(s):  
T Cell ◽  
Epithelial Cells ◽  
Cell Maturation ◽  
Thymic Epithelial Cells ◽  
In Vitro System ◽  
T Cell Maturation

scholarly journals Foxa1 and Foxa2 in thymic epithelial cells (TEC) regulate medullary TEC and regulatory T-cell maturation

2018 ◽  
Vol 93 ◽  
pp. 131-138 ◽  
Author(s):  
Ching-In Lau ◽  
Diana C. Yánez ◽  
Anisha Solanki ◽  
Eleftheria Papaioannou ◽  
José Ignacio Saldaña ◽  
...  
Keyword(s):  
T Cell ◽  
Epithelial Cells ◽  
Regulatory T Cell ◽  
Cell Maturation ◽  
Thymic Epithelial Cells ◽  
T Cell Maturation

Pax1 is expressed during development of the thymus epithelium and is required for normal T-cell maturation

Development ◽  
1996 ◽  
Vol 122 (1) ◽  
pp. 23-30 ◽  
Author(s):  
J. Wallin ◽  
H. Eibel ◽  
A. Neubuser ◽  
J. Wilting ◽  
H. Koseki ◽  
...  
Keyword(s):  
T Cell ◽  
Regulatory Protein ◽  
Cell Maturation ◽  
Thymic Epithelial Cells ◽  
Total Size ◽  
Thymic Development ◽  
Transcriptional Regulatory ◽  
Thymus Cells ◽  
T Cell Maturation ◽  
Third Pharyngeal Pouch

Pax1 is a transcriptional regulatory protein expressed during mouse embryogenesis and has been shown to have an important function in vertebral column development. Expression of Pax1 mRNA in the embryonic thymus has been reported previously. Here we show that Pax1 protein expression in thymic epithelial cells can be detected throughout thymic development and in the adult. Expression starts in the early endodermal epithelium lining the foregut region and includes the epithelium of the third pharyngeal pouch, a structure giving rise to part of the thymus epithelium. In early stages of thymus development a large proportion of thymus cells expresses Pax1. With increasing age, the proportion of Pax1-expressing cells is reduced and in the adult mouse only a small fraction of cortical thymic stromal cells retains strong Pax1 expression. Expression of Pax1 in thymus epithelium is necessary for establishing the thymus microenvironment required for normal T cell maturation. Mutations in the Pax-1 gene in undulated mice affect not only the total size of the thymus but also the maturation of thymocytes. The number of thymocytes is reduced about 2- to 5-fold, affecting mainly the CD4+8+ immature and CD4+ mature thymocyte subsets. The expression levels of major thymocyte surface markers remains unchanged with the exception of Thy-1 which was found to be expressed at 3- to 4-fold higher levels.

scholarly journals Inflammatory Cytokines Regulate T-Cell Development from Blood Progenitor Cells in a Stage and Dose-Specifc Manner

2021 ◽  
Author(s):  
John M. Edgar ◽  
Peter W. Zandstra
Keyword(s):  
T Cells ◽  
T Cell ◽  
Progenitor Cells ◽  
T Cell Development ◽  
Cell Development ◽  
Cell Maturation ◽  
Hematopoietic Stem ◽  
Lineage Differentiation ◽  
T Cell Maturation

ABSTRACTT-cell development from hematopoietic stem and progenitor cells (HSPCs) is tightly regulated through Notch pathway activation by the Notch ligands Delta-like (DL) 1 and 4 and Jagged-2. Other molecules, such as stem cell factor (SCF), FMS-like tyrosine kinase 3 ligand (Flt3L) and interleukin (IL)-7, play a supportive role in regulating the survival, differentiation, and proliferation of developing progenitor (pro)T-cells. Numerous other signaling molecules are known to instruct T-lineage development in vivo, but little work has been done to optimize their use for T-cell production in vitro. Using a defined T-lineage differentiation assay consisting of plates coated with the Notch ligand DL4 and adhesion molecule VCAM-1, we performed a cytokine screen that identified IL-3 and tumor necrosis factor α (TNFα) as enhancers of proT-cell differentiation and expansion. Mechanistically, we found that TNFα induced T-lineage differentiation through the positive regulation of T-lineage genes GATA3, TCF7, and BCL11b. TNFα also synergized with IL-3 to induce proliferation by upregulating the expression of the IL-3 receptor on CD34+ HSPCs, yielding 753.2 (532.4-1026.9; 5-95 percentile)-fold expansion of total cells after 14 days compared to 8.9 (4.3-21.5)-fold expansion in conditions without IL-3 and TNFα. We then optimized cytokine concentrations for T-cell maturation. Focusing on T-cell maturation, we used quantitative models to optimize dynamically changing cytokine requirements and used these to construct a three-stage assay for generating CD3+CD4+CD8+ and CD3+CD4−CD8+ T-cells. Our work provides new insight into T-cell development and a robust in vitro assay for generating T-cells to enable clinical therapies for treating cancer and immune disorders.

scholarly journals Doxycycline Protects Thymic Epithelial Cells from Mitomycin C-Mediated Apoptosis In Vitro via Trx2-NF-κB-Bcl-2/Bax Axis

2016 ◽  
Vol 38 (2) ◽  
pp. 449-460 ◽  
Author(s):  
Jun Wang ◽  
Ya Zhuo ◽  
Lei Yin ◽  
Hui Wang ◽  
Yanqiu Jiang ◽  
...  
Keyword(s):  
T Cell ◽  
Epithelial Cells ◽  
Mitomycin C ◽  
Mapk Signaling ◽  
Thymic Epithelial Cells ◽  
Cell Repertoire ◽  
Gm Csf ◽  
Induced Apoptosis ◽  
A Tec

Background/Aims: Age-associated and stress-induced involution of the thymus is accompanied by reduced numbers of thymic epithelial cells (TECs) and severe reduction in peripheral T cell repertoire specificities. These events seriously affect immune function, but the mechanisms involved are unclear. Our preliminary findings showed that doxycycline (Dox) could drive the proliferation of a TEC line (MTEC1 cells) partially via the MAPK signaling pathway. Dox can also up-regulate IL-6 and GM-CSF expression via the NF-κB and MAPK/ERK pathways. Herein, we investigate the effects and mechanisms used by Dox that protect against mitomycin C (MMC)-induced MTEC1 cell apoptosis. Methods: MTEC1 cells were treated with Dox, MMC, and Dox plus MMC for different amounts of time. The expression of Trx2, NF-κB, Bcl-2, and Bax proteins were then detected by western blotting. Results: Our findings show that Dox protects MTEC1 cells from MMC-induced apoptosis. Dox up-regulated the expression of Trx2 and promoted NF-κB phosphorylation. Meanwhile, Dox also increased the expression of Bcl-2, partially reduced the expression of Bax, and normalized the ratio of Bcl-2 to Bax. Conclusion: Dox exerts an anti-apoptosis function via the NF-κB-Bcl-2/Bax and Trx2-ASK1/JNK pathways in vitro. Therefore, Dox may represent a drug that could be used to attenuate thymic senescence, rescue thymic function, and promote T cell reconstitution.

In Vitro T Cell Differentiation from Human Hematopoietic Stem Cells (HSC) and Lymphoid Progenitors.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4159-4159
Author(s):  
Qian-Lin Hao ◽  
Yuhua Zhu ◽  
Lora Barsky ◽  
Ewa Zielinska ◽  
Mary A. Price ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
T Cell ◽  
Nk Cells ◽  
Culture System ◽  
Cell Maturation ◽  
Cell Production ◽  
Cell Commitment ◽  
T Cell Maturation

Abstract The study of human thymopoiesis has been hampered by our inability to provide all the signals of the normal thymic microenvironment to human stem and progenitor cells in vitro. Although T cell production from CD34+ progenitors can be achieved in vitro, purified populations of the immature CD34+CD38- cells (which include HSC) do not generate T cells reliably with the existing assays. The most commonly cited system for human T cell assay, Fetal thymic organ culture (FTOC) from immune deficient mice, is logistically very difficult and low seeding efficiency precludes its use for clonal assays. Recently, a murine bone marrow stromal line engineered to constitutively express Delta-1 ligand (OP9-DL1 stroma), has been shown to support production of mature murine T cells from HSC [Schmitt, 2002] and ES cells [Schmitt, 2004]. Here we report the use of modifications to the OP9-DL1 system that allow assay of human T lymphopoiesis from HSC and progenitors. CD34+lin-CD38- cells (HSC) from cord blood (CB) and bone marrow (BM)cultured on OP9 control stroma (that does not express DL) produced CD19+ B cells and CD56+ NK cells and maintained CD34+ cells for several weeks. However, T cell commitment was not evident in these control cultures. In contrast, culture of HSC on OP9-DL1 stroma (Dr. Zuniga-Pflucker) resulted in robust growth of T cell precursors (as shown by expression of CD7 and cytoplasmic (cy) CD3 in over 50% of cells) and mRNA for pre-Tα. NK cells were also increased on OP9-DL1 relative to control OP9 stroma but B cell production was lost. Although T cell commitment to the CD34+CD7+ and CD34-cyCD3+CD7+ cells stages was accomplished on OP9-DL1 stroma, markers of further T cell maturation such as surface CD3, CD4 and CD8, were absent on the cells produced. Multiple combinations of growth factors (IL-7, IL-2, IL-15, IL-3, ckit ligand (KL) and thrombopoietin (Tpo)) to the OP9-DL1 stromal cultures failed to achieve further T cell maturation, although the combination of KL, Tpo, IL-7 produced optimal cell growth. However, when conditioned medium collected from human thymic stromal cultures (thyCM) (medium collected from adherent cells derived from human postnatal thymus and filtered through 0.45u) was added to OP9-DL1 co-cultures, cyCD3+ cells were increased and differentiation of CD3+CD4+ cells was achieved. Thymic CD34+lin-CD7+ cells produced large numbers of CD3+CD4+CD8+, CD4+CD8- and CD4-CD8+ cells in this system. We have now used the OP9-DL1/thyCM system to demonstrate the T cell potential of CB and BM CD34+CD38-lin-CD7- (HSC) and CB CD34+CD38-lin-CD7+ (CLP). In addition, efficient cloning of single CB HSC and primitive thymic progenitors has been achieved using this culture system. The data suggests that, although DL-1 is required and sufficient for early T lymphoid commitment, other factor(s) specific to human thymic stroma are required for production of more mature T cells; these factors can now be identified using the OP9-DL1 system. The availability of a simple monolayer culture system that allows T lymphoid commitment from HSC and progenitors and further differentiation to mature T cells will be of great value in the study of the lineage potential of progenitor populations and the regulation of human thymopoiesis.

scholarly journals Interferons Mediate Terminal Differentiation of Human Cortical Thymic Epithelial Cells

2002 ◽  
Vol 76 (13) ◽  
pp. 6415-6424 ◽  
Author(s):  
Pierre-Olivier Vidalain ◽  
David Laine ◽  
Yona Zaffran ◽  
Olga Azocar ◽  
Christine Servet-Delprat ◽  
...  
Keyword(s):  
T Cell ◽  
Epithelial Cells ◽  
Measles Virus ◽  
Present Report ◽  
Thymic Epithelial Cells ◽  
Thymic Epithelial Cell ◽  
Induced Differentiation ◽  
Thymic Epithelium ◽  
Thymus Epithelial Cells

ABSTRACT In the thymus, epithelial cells comprise a heterogeneous population required for the generation of functional T lymphocytes, suggesting that thymic epithelium disruption by viruses may compromise T-cell lymphopoiesis in this organ. In a previous report, we demonstrated that in vitro, measles virus induced differentiation of cortical thymic epithelial cells as characterized by (i) cell growth arrest, (ii) morphological and phenotypic changes, and (iii) apoptotis as a final step of this process. In the present report, we have analyzed the mechanisms involved. First, measles virus-induced differentiation of thymic epithelial cells is shown to be strictly dependent on beta interferon (IFN-β) secretion. In addition, transfection with double-stranded RNA, a common intermediate of replication for a broad spectrum of viruses, is reported to similarly mediate thymic epithelial cell differentiation through IFN-β induction. Finally, we demonstrated that recombinant IFN-α, IFN-β, or IFN-γ was sufficient to induce differentiation and apoptosis of uninfected thymic epithelial cells. These observations suggested that interferon secretion by either infected cells or activated leukocytes, such as plasmacytoid dendritic cells or lymphocytes, may induce thymic epithelium disruption in a pathological context. Thus, we have identified a new mechanism that may contribute to thymic atrophy and altered T-cell lymphopoiesis associated with many infections.

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