Peptide-amidating Enzymes Are Expressed in the Stellate Epithelial Cells of the Thymic Medulla

1998 ◽  
Vol 46 (5) ◽  
pp. 661-668 ◽  
Author(s):  
Alfredo Martínez ◽  
Andrew Farr ◽  
Michele D. Vos ◽  
Frank Cuttitta ◽  
Anthony M. Treston
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Cell Lines ◽  
B Lymphocyte ◽  
Regulatory Peptides ◽  
Convincing Evidence ◽  
Thymic Epithelial Cells ◽  
Thymic Epithelial Cell ◽  
Epithelial Cell Lines ◽  
Amidating Enzymes

C-terminal amidation is a post-translational processing step necessary to convey biological activity to a large number of regulatory peptides. In this study we have demonstrated that the peptidyl-glycine α-amidating monooxygenase enzyme complex (PAM) responsible for this activity is located in the medullary stellate epithelial cells of the thymus and in cultured epithelial cells bearing a medullary phenotype, using Northern blot, immunocytochemistry, in situ hybridization, and enzyme assays. Immunocytochemical localization revealed a granular pattern in the cytoplasm of the stellate cells, which were also positive for cytokeratins and a B-lymphocyte-associated antigen. The presence of PAM activity in medium conditioned by thymic epithelial cell lines suggests that PAM is a secreted product of these cells. Among the four epithelial cell lines examined, there was a direct correlation between PAM activity and content of oxytocin, an amidated peptide. Taken together, these data provide convincing evidence that thymic epithelial cells have the capacity to generate amidated peptides that may influence T-cell differentiation and suggest that the amidating enzymes could play an important role in the regulation of thymic physiology.

scholarly journals High efficiency transfection of thymic epithelial cell lines and primary thymic epithelial cells by Nucleofection.

2015 ◽  
Author(s):  
Richard T O'Neil ◽  
Qiaozhi Wei ◽  
Brian G Condie
Keyword(s):  
T Cells ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Cell Lines ◽  
High Efficiency ◽  
Thymic Epithelial Cells ◽  
Thymic Epithelial Cell ◽  
Thymus Tissue ◽  
Primary Mouse ◽  
Epithelial Cell Lines

Thymic epithelial cells (TECs) are required for the development and differentiation of T cells and are sufficient for the positive and negative selection of developing T cells. Although TECs play a critical role in T cell biology, simple, efficient and readily scalable methods for the transfection of TEC lines and primary TECs have not been described. We tested the efficiency of Nucleofection for the transfection of 4 different mouse thymic epithelial cell lines that had been derived from cortical or medullary epithelium. We also tested primary mouse thymic epithelial cells isolated from fetal and postnatal stages. We found that Nucleofection was highly efficient for the transfection of thymic epithelial cells, with transfection efficiencies of 30-70% for the cell lines and 15-35% for primary TECs with low amounts of cell death. Efficient transfection by Nucleofection can be performed with established cortical and medullary thymic epithelial cell lines as well as primary TECs isolated from E15.5 day fetal thymus or postnatal day 3 or 30 thymus tissue. The high efficiency of Nucleofection for TEC transfection will enable the use of TEC lines in high throughput transfection studies and simplifies the transfection of primary TECs for in vitro or in vivo analysis.

A novel form of cellular communication among thymic epithelial cells: intercellular calcium wave propagation

2003 ◽  
Vol 285 (5) ◽  
pp. C1304-C1313 ◽  
Author(s):  
O. K. Nihei ◽  
A. C. Campos de Carvalho ◽  
D. C. Spray ◽  
W. Savino ◽  
L. A. Alves
Keyword(s):  
Wave Propagation ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Gap Junctions ◽  
Thymic Epithelial Cells ◽  
Calcium Wave ◽  
Thymic Epithelial Cell ◽  
Calcium Waves ◽  
Thymic Nurse Cells ◽  
Epithelial Cell Lines

We here describe intercellular calcium waves as a novel form of cellular communication among thymic epithelial cells. We first characterized the mechanical induction of intercellular calcium waves in different thymic epithelial cell preparations: cortical 1-4C18 and medullary 3-10 thymic epithelial cell lines and primary cultures of thymic “nurse” cells. All thymic epithelial preparations responded with intercellular calcium wave propagation after mechanical stimulation. In general, the propagation efficacy of intercellular calcium waves in these cells was high, reaching 80-100% of the cells within a given confocal microscopic field, with a mean velocity of 6-10 μm/s and mean amplitude of 1.4- to 1.7-fold the basal calcium level. As evaluated by heptanol and suramin treatment, our results suggest the participation of both gap junctions and P2 receptors in the propagation of intercellular calcium waves in thymic nurse cells and the more prominent participation of gap junctions in thymic epithelial cell lines. Finally, in cocultures, the transmission of intercellular calcium wave was not observed between the mechanically stimulated thymic epithelial cell and adherent thymocytes, suggesting that intercellular calcium wave propagation is limited to thymic epithelial cells and does not affect the neighboring thymocytes. In conclusion, these data describe for the first time intercellular calcium waves in thymic epithelial cells and the participation of both gap junctions and P2 receptors in their propagation.

scholarly journals Medullary but not cortical thymic epithelial cells present soluble antigens to helper T cells.

1992 ◽  
Vol 175 (6) ◽  
pp. 1601-1605 ◽  
Author(s):  
T Mizuochi ◽  
M Kasai ◽  
T Kokuho ◽  
T Kakiuchi ◽  
K Hirokawa
Keyword(s):  
Monoclonal Antibody ◽  
Epithelial Cells ◽  
Cell Lines ◽  
Helper T Cells ◽  
Thymic Epithelial Cells ◽  
Thymic Epithelial Cell ◽  
Functional Difference ◽  
Epithelial Cell Lines ◽  
T Cell Lines ◽  
Antigen Presenting

Thymic epithelial cell lines (TECs) were established from newborn C57BL/6 mice. They were classified into two types (medullary and cortical TECs) by using the monoclonal antibody (Th-3) that recognizes the meshwork structure of thymic cortical epithelial cells. Antigen-presenting activity of each TEC was determined by using ovalbumin-specific, I-Ab-restricted helper T cell lines. It was demonstrated that the medullary but not the cortical TECs functioned as antigen-presenting cells. This is the first evidence for the functional difference between the cortical and the medullary TEC.

CD10 is expressed on human thymic epithelial cell lines and modulates thymopentin‐induced cell proliferation

1997 ◽  
Vol 11 (12) ◽  
pp. 1003-1011 ◽  
Author(s):  
Sandrine Guérin ◽  
Bernard Mari ◽  
Edgar Fernandez ◽  
Nathalie Belhacene ◽  
Maria Luisa Toribio ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Epithelial Cell ◽  
Cell Lines ◽  
Thymic Epithelial Cell ◽  
Epithelial Cell Lines

scholarly journals Urine-Derived Epithelial Cell Lines: A New Tool to Model Fragile X Syndrome (FXS)

Cells ◽  
2020 ◽  
Vol 9 (10) ◽  
pp. 2240
Author(s):  
Marwa Zafarullah ◽  
Mittal Jasoliya ◽  
Flora Tassone
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Fragile X Syndrome ◽  
Cell Lines ◽  
Behavioral Problems ◽  
Cell Model ◽  
Fragile X ◽  
Healthy Controls ◽  
Full Mutation ◽  
Epithelial Cell Lines

Fragile X syndrome (FXS) is an X-linked neurodevelopmental condition associated with intellectual disability and behavioral problems due to the lack of the Fragile X mental retardation protein (FMRP), which plays a crucial role in synaptic plasticity and memory. A desirable in vitro cell model to study FXS would be one that can be generated by simple isolation and culture method from a collection of a non-invasive donor specimen. Currently, the various donor-specific cells can be isolated mainly from peripheral blood and skin biopsy. However, they are somewhat invasive methods for establishing cell lines from the primary subject material. In this study, we characterized a cost-effective and straightforward method to derive epithelial cell lines from urine samples collected from participants with FXS and healthy controls (TD). The urine-derived cells expressed epithelial cell surface markers via fluorescence-activated cell sorting (FACS). We observed inter, and the intra-tissue CGG mosaicism in the PBMCs and the urine-derived cells from participants with FXS potentially related to the observed variations in the phenotypic and clinical presentation FXS. We characterized these urine-derived epithelial cells for FMR1 mRNA and FMRP expression and observed some expression in the lines derived from full mutation mosaic participants. Further, FMRP expression was localized in the cytoplasm of the urine-derived epithelial cells of healthy controls. Deficient FMRP expression was also observed in mosaic males, while, as expected, no expression was observed in cells derived from participants with a hypermethylated full mutation.

Mouse thymic epithelial cell lines expressing “Aire” and peripheral tissue-specific antigens reproduce in vitro negative selection of T cells

2011 ◽  
Vol 317 (14) ◽  
pp. 2019-2030 ◽  
Author(s):  
Yoshitaka Yamaguchi ◽  
Atsushi Takayanagi ◽  
Jiabing Chen ◽  
Kosuke Sakai ◽  
Jun Kudoh ◽  
...  
Keyword(s):  
T Cells ◽  
Epithelial Cell ◽  
Cell Lines ◽  
Negative Selection ◽  
Thymic Epithelial Cell ◽  
Tissue Specific ◽  
Epithelial Cell Lines ◽  
Specific Antigens ◽  
Selection Of

scholarly journals Differential Responses by Human Respiratory Epithelial Cell Lines to Respiratory Syncytial Virus Reflect Distinct Patterns of Infection Control

2018 ◽  
Vol 92 (15) ◽  
Author(s):  
Philippa Hillyer ◽  
Rachel Shepard ◽  
Megan Uehling ◽  
Mina Krenz ◽  
Faruk Sheikh ◽  
...  
Keyword(s):  
Respiratory Syncytial Virus ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Cell Lines ◽  
A549 Cells ◽  
Type I ◽  
Epithelial Cell Lines ◽  
Rsv Infection ◽  
Syncytial Virus ◽  
Respiratory Epithelial

ABSTRACT Respiratory syncytial virus (RSV) infects small foci of respiratory epithelial cells via infected droplets. Infection induces expression of type I and III interferons (IFNs) and proinflammatory cytokines, the balance of which may restrict viral replication and affect disease severity. We explored this balance by infecting two respiratory epithelial cell lines with low doses of recombinant RSV expressing green fluorescent protein (rgRSV). A549 cells were highly permissive, whereas BEAS-2B cells restricted infection to individual cells or small foci. After infection, A549 cells expressed higher levels of IFN-β-, IFN-λ-, and NF-κB-inducible proinflammatory cytokines. In contrast, BEAS-2B cells expressed higher levels of antiviral interferon-stimulated genes, pattern recognition receptors, and other signaling intermediaries constitutively and after infection. Transcriptome analysis revealed that constitutive expression of antiviral and proinflammatory genes predicted responses by each cell line. These two cell lines provide a model for elucidating critical mediators of local control of viral infection in respiratory epithelial cells. IMPORTANCE Airway epithelium is both the primary target of and the first defense against respiratory syncytial virus (RSV). Whether RSV replicates and spreads to adjacent epithelial cells depends on the quality of their innate immune responses. A549 and BEAS-2B are alveolar and bronchial epithelial cell lines, respectively, that are often used to study RSV infection. We show that A549 cells are permissive to RSV infection and express genes characteristic of a proinflammatory response. In contrast, BEAS-2B cells restrict infection and express genes characteristic of an antiviral response associated with expression of type I and III interferons. Transcriptome analysis of constitutive gene expression revealed patterns that may predict the response of each cell line to infection. This study suggests that restrictive and permissive cell lines may provide a model for identifying critical mediators of local control of infection and stresses the importance of the constitutive antiviral state for the response to viral challenge.

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