scholarly journals Aire controls the differentiation program of thymic epithelial cells in the medulla for the establishment of self-tolerance

2008 ◽  
Vol 205 (12) ◽  
pp. 2827-2838 ◽  
Author(s):  
Masashi Yano ◽  
Noriyuki Kuroda ◽  
Hongwei Han ◽  
Makiko Meguro-Horike ◽  
Yumiko Nishikawa ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Fluorescent Protein ◽  
Morphological Changes ◽  
Cell Types ◽  
Salivary Protein ◽  
Epidermal Differentiation ◽  
Thymic Epithelial Cells ◽  
Reactive Protein ◽  
Medullary Thymic Epithelial Cells ◽  
Green Fluorescent

The roles of autoimmune regulator (Aire) in the expression of the diverse arrays of tissue-restricted antigen (TRA) genes from thymic epithelial cells in the medulla (medullary thymic epithelial cells [mTECs]) and in organization of the thymic microenvironment are enigmatic. We approached this issue by creating a mouse strain in which the coding sequence of green fluorescent protein (GFP) was inserted into the Aire locus in a manner allowing concomitant disruption of functional Aire protein expression. We found that Aire+ (i.e., GFP+) mTECs were the major cell types responsible for the expression of Aire-dependent TRA genes such as insulin 2 and salivary protein 1, whereas Aire-independent TRA genes such as C-reactive protein and glutamate decarboxylase 67 were expressed from both Aire+ and Aire− mTECs. Remarkably, absence of Aire from mTECs caused morphological changes together with altered distribution of mTECs committed to Aire expression. Furthermore, we found that the numbers of mTECs that express involucrin, a marker for terminal epidermal differentiation, were reduced in Aire-deficient mouse thymus, which was associated with nearly an absence of Hassall's corpuscle-like structures in the medulla. Our results suggest that Aire controls the differentiation program of mTECs, thereby organizing the global mTEC integrity that enables TRA expression from terminally differentiated mTECs in the thymic microenvironment.

scholarly journals AIRE Recruits P-TEFb for Transcriptional Elongation of Target Genes in Medullary Thymic Epithelial Cells

2007 ◽  
Vol 27 (24) ◽  
pp. 8815-8823 ◽  
Author(s):  
Irena Oven ◽  
Naděžda Brdičková ◽  
Jiri Kohoutek ◽  
Tomaž Vaupotič ◽  
Mojca Narat ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Target Genes ◽  
Ectopic Expression ◽  
Lymphocytic Infiltration ◽  
Salivary Protein ◽  
Thymic Epithelial Cells ◽  
Transcriptional Elongation ◽  
Medullary Thymic Epithelial Cells

ABSTRACT AIRE is a transcriptional activator that directs the ectopic expression of many tissue-specific genes in medullary thymic epithelial cells, which plays an important role in the negative selection of autoreactive T cells. However, its mechanism of action remains poorly understood. In this study, we found that AIRE regulates the step of elongation rather than initiation of RNA polymerase II. For these effects, AIRE bound and recruited P-TEFb to target promoters in medullary thymic epithelial cells. In these cells, AIRE activated the ectopic transcription of insulin and salivary protein 1 genes. Indeed, by chromatin immunoprecipitation, we found that RNA polymerase II was already engaged on these promoters but was unable to elongate in the absence of AIRE. Moreover, the genetic inactivation of cyclin T1 from P-TEFb abolished the transcription of AIRE-responsive genes and led to lymphocytic infiltration of lacrimal and salivary glands in the CycT1−/− mouse. Our findings reveal critical steps by which AIRE regulates the transcription of genes that control central tolerance in the thymus.

scholarly journals Stage-selective differential expression of eukaryotic initiation factor (Eif) genes in the medullary epithelial cells of the murine thymus.

2020 ◽  
Author(s):  
Shahan Mamoor
Keyword(s):  
Epithelial Cells ◽  
Adult Development ◽  
Microarray Dataset ◽  
Cell Types ◽  
Initiation Factor ◽  
Thymic Epithelial Cells ◽  
Initiation Factors ◽  
Eukaryotic Initiation Factors ◽  
Medullary Thymic Epithelial Cells ◽  
Differential Gene

The thymus is the site of positive and negative selection in mammals (1), the process by which developing lymphocytes are selected for or deleted based on their ability to encounter foreign or self antigen, respectively, with optimal binding affinity (2, 3). The thymus consists of a medulla and a cortex , with mTEC, or medullary thymic epithelial cells expressing high or low amounts of the class II major histocompatibility complex (mTEChi and mTEClo) (4, 5). To understand how the mTEC transcriptome changes over mammalian adult development, we performed global differential gene expression profiling using a public microarray dataset to compare the transcriptomes of mTEC at months 1, 3, and 6 (6). We found that multiple eukaryotic initiation factors, or Eif genes, were differentially expressed by mTEClo and mTEChi. These data suggest a unique requirement for specific eukaryotic initiation factors during translation of proteins in the mTEClo and mTEChi cell types of the thymus.

EGFP expression in goat mammary epithelial cells

2006 ◽  
Vol 3 (1) ◽  
pp. 71-74 ◽  
Author(s):  
Zheng Yue-Mao ◽  
An Zhi-Xing ◽  
Peng Xin-Rong ◽  
Shi Yu-Qiang ◽  
Zhang Yong
Keyword(s):  
Epithelial Cells ◽  
Fluorescent Protein ◽  
Mammary Epithelial Cells ◽  
Cell Types ◽  
Mammary Epithelial ◽  
Egfp Expression ◽  
Green Fluorescent ◽  
Mammary Gland Cells ◽  
Different Cell Types ◽  
Goat Mammary Epithelial Cells

AbstractPrimary culture of goat mammary gland cells was achieved by outgrowth of migrating cells from fragments of tissue. The fibroblast and epithelial cells were purified according to their different sensitivity to trypsin and the characteristics of goat mammary epithelial cells were observed under a light microscope. The results showed that the purified goat mammary epithelial cells retained normal characteristics until the 15th passage. The purified mammary epithelial cells propagated and formed a dome-like structure that resembled a nipple and was called a ‘milk orb’. The mammary epithelial cells could produce and secrete milk. There were different cell types: the majority were short shuttle-like or polygons, resembling a beehive; while some were large, flat and round; and others were elongated. The enhanced green fluorescent protein (EGFP) gene was transferred successfully into the goat mammary epithelial cells using electrotransfection, and its expression was observed under a fluoroscope.

scholarly journals Lymphotoxin β Receptor Regulates the Development of CCL21-Expressing Subset of Postnatal Medullary Thymic Epithelial Cells

2013 ◽  
Vol 190 (10) ◽  
pp. 5110-5117 ◽  
Author(s):  
Enkhsaikhan Lkhagvasuren ◽  
Mie Sakata ◽  
Izumi Ohigashi ◽  
Yousuke Takahama
Keyword(s):  
Epithelial Cells ◽  
Thymic Epithelial Cells ◽  
Medullary Thymic Epithelial Cells ◽  
Β Receptor

scholarly journals Impact of Heterogeneity within Cultured Cells on Bacterial Invasion: Analysis of Pseudomonas aeruginosa andSalmonella enterica Serovar Typhi Entry into MDCK cells by Using a Green Fluorescent Protein-Labelled Cystic Fibrosis Transmembrane Conductance Regulator Receptor

2000 ◽  
Vol 68 (2) ◽  
pp. 861-870 ◽  
Author(s):  
A. Alev Gerçeker ◽  
Tanweer Zaidi ◽  
Peter Marks ◽  
David E. Golan ◽  
Gerald B. Pier
Keyword(s):  
Epithelial Cells ◽  
Protein Expression ◽  
Fluorescent Protein ◽  
Cultured Cells ◽  
Mdck Cells ◽  
Transmembrane Conductance Regulator ◽  
Transmembrane Conductance ◽  
Bacterial Uptake ◽  
Cftr Protein ◽  
Green Fluorescent

ABSTRACT The cystic fibrosis transmembrane conductance regulator (CFTR) is a chloride ion channel that also serves as a receptor for entry ofPseudomonas aeruginosa and Salmonella entericaserovar Typhi into epithelial cells. To evaluate heterogeneity in CFTR protein expression in cultured cells and the effect of heterogeneity on internalization of different P. aeruginosa and serovar Typhi strains, we used two-color flow cytometry and confocal laser microscopy to study bacterial uptake by Madin-Darby canine kidney (MDCK) type I epithelial cells stably expressing a green fluorescent protein (GFP)-CFTR fusion construct (MDCK–GFP-CFTR cells). We found a strong correlation between cell size and GFP-CFTR protein expression, with 60 to 70% of cells expressing low levels of GFP-CFTR protein, 20 to 30% expressing intermediate levels, and <10% expressing high levels. The cells were sorted into low-, intermediate-, or high-level producers of CFTR protein; in vitro growth of each sorted population yielded the same distribution of CFTR protein expression as that in the original population. Cells expressing either low or high levels of CFTR protein internalized bacteria poorly; maximal bacterial uptake occurred in the cells expressing intermediate levels of CFTR protein. Treatment of MDCK cells with sodium butyrate markedly enhanced the production of CFTR protein without increasing cell size; butyrate treatment also increased the proportion of cells with internalized bacteria. However, there were fewer bacteria per butyrate-treated cell and, for P. aeruginosa, there was an overall decrease in the total level of bacterial uptake. The most highly ingested bacterial strains were internalized by fewer total MDCK–GFP-CFTR cells, indicating preferential bacterial uptake by a minority of epithelial cells within a given culture. Confocal fluorescence microscopy showed that P. aeruginosa and serovar Typhi induced cytoplasmic accumulation of CFTR protein close to the plasma membrane where the bacteria were adherent. These results show that within a population of MDCK–GFP-CFTR cells, there are cells with markedly different abilities to ingest bacteria via CFTR, the majority of the P. aeruginosa and serovar Typhi cells are ingested by the one-fourth to one-third of the cells that exhibit an intermediate size and level of CFTR protein expression, and overexpression of the CFTR receptor does not increase total bacterial uptake but rather allows more epithelial cells to ingest fewer total bacteria.

scholarly journals Foxn1 Regulates Lineage Progression in Cortical and Medullary Thymic Epithelial Cells But Is Dispensable for Medullary Sublineage Divergence

PLoS Genetics ◽  
2011 ◽  
Vol 7 (11) ◽  
pp. e1002348 ◽  
Author(s):  
Craig S. Nowell ◽  
Nicholas Bredenkamp ◽  
Stéphanie Tetélin ◽  
Xin Jin ◽  
Christin Tischner ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Thymic Epithelial Cells ◽  
Medullary Thymic Epithelial Cells

Developmental derivation of vocal fold mucosa from human induced pluripotent stem cells

2019 ◽  
Author(s):  
Vlasta Lungova ◽  
Susan Thibeault
Keyword(s):  
Epithelial Cells ◽  
Pluripotent Stem Cells ◽  
Vocal Fold ◽  
Fluorescent Protein ◽  
Three Dimensional ◽  
Stratified Squamous Epithelium ◽  
Three Dimensional Models ◽  
Induced Pluripotent ◽  
Green Fluorescent

Abstract Development of treatments for vocal dysphonia has been inhibited by lack of human vocal fold (VF) mucosa models because of difficulty in procuring VF epithelial cells, epithelial cells’ limited proliferative capacity and absence of cell lines. We report development of engineered VF mucosae from hiPSC, transfected via TALEN constructs for green fluorescent protein, that mimic development of VF epithelial cells in utero. Modulation of FGF signaling achieves stratified squamous epithelium from definitive and anterior foregut derived cultures. Robust culturing of these cells on collagen-fibroblast constructs produces three-dimensional models comparable to in vivo VF mucosa.

scholarly journals Diversity in medullary thymic epithelial cells controls the activity and availability of iNKT cells

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Beth Lucas ◽  
Andrea J. White ◽  
Emilie J. Cosway ◽  
Sonia M. Parnell ◽  
Kieran D. James ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Thymic Epithelial Cells ◽  
Inkt Cells ◽  
Medullary Thymic Epithelial Cells
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