Regional differences in glycoconjugates of intestinal M cells in mice: potential targets for mucosal vaccines

1994 ◽  
Vol 267 (6) ◽  
pp. G1108-G1121 ◽  
Author(s):  
P. J. Giannasca ◽  
K. T. Giannasca ◽  
P. Falk ◽  
J. I. Gordon ◽  
M. R. Neutra
Keyword(s):  
Early Stage ◽  
Basolateral Membrane ◽  
M Cells ◽  
Type I ◽  
M Cell ◽  
Ulex Europaeus ◽  
Terminal Galactose ◽  
Mucosal Explants ◽  
Endocytic Vesicles

We have used a panel of lectins and antibodies to describe the composition of complex carbohydrates associated with M cells in various regions of the intestinal tract of adult BALB/c mice. The fucose-specific lectin Ulex europaeus agglutinin type I (UEA I) is a marker of M cells in the small intestine and recognized M cells at an early stage of differentiation. Subpopulations of M cells in a single follicle-associated epithelium (FAE) could be distinguished by different fucose-specific probes. Certain lectins revealed that M cells have basal processes that extend into the underlying lymphoid tissue. Colonic and rectal M cells display glycosylation patterns distinct from M cells of Peyer's patches and are characterized by terminal galactose. UEA I selectively adhered to Peyer's patch M cells in mucosal explants and in ligated intestinal loops in vivo. The lectin was taken up into endocytic vesicles and transported to the intra-epithelial pocket and other domains of the basolateral membrane. Thus M cell-specific glycoconjugates could serve as "receptors" for targeting of lectin-antigen conjugates to the mucosal immune system.

scholarly journals Identifying human and murine M cells in vitro

2019 ◽  
Vol 244 (7) ◽  
pp. 554-564 ◽  
Author(s):  
Ana Klisuric ◽  
Benjamin Thierry ◽  
Ludivine Delon ◽  
Clive A Prestidge ◽  
Rachel J Gibson
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
In Vitro Models ◽  
Oral Drug ◽  
M Cells ◽  
Cell Marker ◽  
M Cell ◽  
Follicle Associated Epithelium

M cells are an epithelial cell population found in the follicle-associated epithelium overlying gut-associated lymphoid tissues. They are specialized in the transcytosis of luminal antigens. Their transcytotic capacity and location in an immunocompetent environment has prompted the study of these cells as possible targets for oral drug delivery systems. Currently, the models most commonly used to study M cells are restricted to in vivo experiments conducted in mice, and in vitro studies conducted in models comprised either of primary epithelial cells or established cell lines of murine or human origin. In vitro models of the follicle-associated epithelium can be constructed in several ways. Small intestinal Lgr5+ stem cells can be cultured into a 3D organoid structure where M cells are induced with RANKL administration. Additionally, in vitro models containing an “M cell-like” population can be obtained through co-culturing intestinal epithelial cells with cells of lymphocytic origin to induce the M cell phenotype. The evaluation of the efficiency of the variations of these models and their relevance to the in vivo human system is hampered by the lack of a universal M cell marker. This issue has also hindered the advancement of M cell-specific targeting approaches aimed at improving the bioavailability of orally administered compounds. This critical review discusses the different approaches utilized in the literature to identify M cells, their efficiency, reliability and relevance, in the context of commonly used models of the follicle-associated epithelium. The outcome of this review is a clearly defined and universally recognized criteria for the assessment of the relevance of models of the follicle-associated models currently used. Impact statement The study of M cells, a specialized epithelial cell type found in the follicle-associated epithelium, is hampered by the lack of a universal M cell marker. As such, many studies lack reliable and universally recognized methods to identify M cells in their proposed models. As a result of this it is difficult to ascertain whether the effects observed are due to the presence of M cells or an unaccounted variable. The outcome of this review is the thorough evaluation of the many M cell markers that have been used in the literature thus far and a proposed criterion for the identification of M cells for future publications. This will hopefully lead to an improvement in the quality of future publications in this field.

scholarly journals Comparative study of transcriptomics-based scoring metrics for the epithelial-hybrid-mesenchymal spectrum

2020 ◽  
Author(s):  
Priyanka Chakraborty ◽  
Jason T George ◽  
Shubham Tripathi ◽  
Herbert Levine ◽  
Mohit Kumar Jolly
Keyword(s):  
Cancer Cells ◽  
Cancer Metastasis ◽  
Epithelial Mesenchymal Transition ◽  
Multinomial Logistic Regression ◽  
M Cells ◽  
M Cell ◽  
Mesenchymal Transition ◽  
Tumor Types

AbstractThe Epithelial-mesenchymal transition (EMT) is a cellular process implicated in embryonic development, wound healing, and pathological conditions such as cancer metastasis and fibrosis. Cancer cells undergoing EMT exhibit enhanced aggressive behavior characterized by drug resistance, tumor-initiation potential, and the ability to evade immune system. Recent in silico, in vitro, and in vivo evidence indicates that EMT is not an all-or-none process; instead, cells stably acquire one or more hybrid epithelial/mesenchymal (E/M) phenotypes which often can be more aggressive than purely epithelial or mesenchymal cell populations. Thus, the EMT status of cancer cells can prove to be a critical estimate of patient prognosis. Recent attempts have employed different transcriptomics signatures to quantify EMT status in cell lines and patient tumors. However, a comprehensive comparison of these methods, including their accuracy in identifying cells in the hybrid E/M phenotype(s), is lacking. Here, we compare three distinct metrics that score EMT on a continuum, based on the transcriptomics signature of individual samples. Our results demonstrate that these methods exhibit good concordance among themselves in quantifying the extent of EMT in a given sample. Moreover, scoring EMT using any of the three methods discerned that cells undergo varying extents of EMT across tumor types. Separately, our analysis also identified tumor types with maximum variability in terms of EMT and associated an enrichment of hybrid E/M signatures in these samples. Moreover, we also found that the multinomial logistic regression (MLR) based metric was capable of distinguishing between ‘pure’ individual hybrid E/M vs. mixtures of epithelial (E) and mesenchymal (M) cells. Our results, thus, suggest that while any of the three methods can indicate a generic trend in the EMT status of a given cell, the MLR method has two additional advantages: a) it uses a small number of predictors to calculate the EMT score, and b) it can predict from the transcriptomic signature of a population whether it is comprised of ‘pure’ hybrid E/M cells at the single-cell level or is instead an ensemble of E and M cell subpopulations.

scholarly journals M-Cell Surface β1 Integrin Expression and Invasin-Mediated Targeting of Yersinia pseudotuberculosis to Mouse Peyer’s Patch M Cells

1998 ◽  
Vol 66 (3) ◽  
pp. 1237-1243 ◽  
Author(s):  
M. Ann Clark ◽  
Barry H. Hirst ◽  
Mark A. Jepson
Keyword(s):  
Cell Surface ◽  
Yersinia Pseudotuberculosis ◽  
M Cells ◽  
Β1 Integrin ◽  
Integrin Expression ◽  
Peyer’S Patch ◽  
Peyer's Patch ◽  
M Cell ◽  
Apical Membranes

ABSTRACT Quantitative analysis of Yersinia pseudotuberculosisinfection of murine gut loops revealed that significantly more wild-type bacteria associated with Peyer’s patch M cells than with dome enterocytes or goblet cells. An invasin-deficient mutant was significantly attenuated for M-cell invasion, while β1 integrin expression was demonstrated in the apical membranes of M cells but not enterocytes. M-cell targeting by Yersinia pseudotuberculosis in vivo may, therefore, be mediated primarily by the interaction of invasin with cell surface β1 integrins.

scholarly journals Mycobacterium avium subsp. paratuberculosis Fibronectin Attachment Protein Facilitates M-Cell Targeting and Invasion through a Fibronectin Bridge with Host Integrins

2004 ◽  
Vol 72 (7) ◽  
pp. 3724-3732 ◽  
Author(s):  
T. E. Secott ◽  
T. L. Lin ◽  
C. C. Wu
Keyword(s):  
Epithelial Cells ◽  
Mycobacterium Avium ◽  
Cell Invasion ◽  
M Cells ◽  
Intestinal Epithelial ◽  
M Cell ◽  
Attachment Protein ◽  
Β3 Integrin ◽  
Mycobacterium Avium Subsp Paratuberculosis

ABSTRACT Efficient attachment and ingestion of Mycobacterium avium subsp. paratuberculosis by cultured epithelial cells requires the expression of a fibronectin (FN) attachment protein homologue (FAP-P) which mediates FN binding by M. avium subsp. paratuberculosis. Invasion of Peyer's patches by M. avium subsp. paratuberculosis occurs through M cells, which, unlike other intestinal epithelial cells, express integrins on their luminal faces. We sought to determine if the interaction between FAP-P of M. avium subsp. paratuberculosis and soluble FN enabled targeting and invasion of M cells by M. avium subsp. paratuberculosis in vivo via these surface integrins. Wild-type and antisense FAP-P mutant M. avium subsp. paratuberculosis strains were injected alone or coinjected with blocking peptides or antibodies into murine gut loops, and immunofluorescence microscopy was performed to assess targeting and invasion of M cells by M. avium subsp. paratuberculosis. Nonopsonized M. avium subsp. paratuberculosis preferentially invaded M cells in murine gut loops. M-cell invasion was enhanced 2.6-fold when M. avium subsp. paratuberculosis was pretreated with FN. Invasion of M cells by the antisense FAP-P mutant of M. avium subsp. paratuberculosis was reduced by 77 to 90% relative to that observed for the control strains. Peptides corresponding to the RGD and synergy site integrin recognition regions of FN blocked M. avium subsp. paratuberculosis invasion of M cells by 75 and 45%, respectively, whereas the connecting segment 1 peptide was noninhibitory. Antibodies against the α5, αV, β1, and β3 integrin subunits inhibited M-cell invasion by 52 to 73%. The results indicate that targeting and invasion of M cells by M. avium subsp. paratuberculosis in vivo is mediated primarily by the formation of an FN bridge formed between FAP-P of M. avium subsp. paratuberculosis and integrins on M cells.

Gene Expressions Induced by Calcium Phosphate Ceramics after Implantation into the Muscle of Rat

2005 ◽  
Vol 475-479 ◽  
pp. 2363-2366
Author(s):  
Jin Rui Xu ◽  
Hong Song Fan ◽  
Yan Fei Tan ◽  
Xing Dong Zhang
Keyword(s):  
Calcium Phosphate ◽  
Collagen Type ◽  
Early Stage ◽  
Collagen Type I ◽  
Type I ◽  
Rt Pcr ◽  
Gene Expressions ◽  
Porous Hydroxyapatite ◽  
Calcium Phosphate Ceramics

The osteoinductivity of calcium phosphate ceramics has been studied extensively, but the mechanism is still unclear and few reports about the molecular mechanism in the osteoinductive process. In this study the osteoblast related gene expressions induced by biomaterials were investigated by isolating the RNA from the tissue grown in porous hydroxyapatite/tricalcium phosphate (HA/TCP) ceramics implanted in rat femur muscle on day 7, 15, 30, 60, 90,120, and analyzed by RT-PCR technique. RNA extracted from muscle without implant was used as control at the same time. The results showed that osteopontin and osteocalcin genes, the important osteoblastic markers, expressed in early stage, on day 7 after implantation, and were detected at any period. Collagen type I gene expressed on day 60, 90 and 120. It revealed that osteoblast differentiation occurred very early before collagen type I expression after implanting HA/TCP ceramics in vivo.

scholarly journals Cell-extracellular matrix interactions in the fluidic phase direct the topology and polarity of self-organized epithelial structures

2020 ◽  
Author(s):  
Mingxing Ouyang ◽  
Jiun-Yann Yu ◽  
Yenyu Chen ◽  
Linhong Deng ◽  
Chin-Lin Guo
Keyword(s):  
Extracellular Matrix ◽  
Large Scale ◽  
Type I Collagen ◽  
Mdck Cells ◽  
Early Stage ◽  
Single Cells ◽  
Confocal Imaging ◽  
Type I

AbstractIn vivo, cells are surrounded by extracellular matrix (ECM). To build organs from single cells, it is generally believed that ECM serves as a large-scale scaffold to coordinate cell positioning and differentiation. Nevertheless, how cells utilize cell-ECM interactions to spatiotemporally coordinate their positioning and differentiation to different ECM at the whole-tissue scale is not fully understood. Here, using in vitro assay with engineered MDCK cells co-expressing H2B-mCherry (nucleus) and gp135 (Podocalyxin)-GFP (apical marker), we show that such spatiotemporal coordination for epithelial morphogenesis and polarization can be initiated and determined by cell-soluble ECM interaction in the fluidic phase. The coordination depends on the native topology of ECM components such as sheet-like basement membrane (BM, mimicked by Matrigel in experiments) and linear fiber-like type I collagen (COL). Two types of coordination are found: scaffold formed by BM (COL) facilitates a close-ended (open-ended) coordination that leads to the formation of lobular (tubular) epithelium, where polarity is preserved throughout the entire lobule/tubule. During lobular formation with BM, polarization of individual cells within the same cluster occurs almost simultaneously, whereas the apicobasal polarization in the presence of COL can start at local regions and proceed in a collective way along the axis of tubule, which might suggest existence of intercellular communications at the cell-population level. Further, in the fluidic phase, we found that cells can form apicobasal polarity throughout the entire lobule/tubule without a complete coverage of ECM at the basal side. Based on reconstructions from time-lapse confocal imaging, this is likely derived from polarization occurring at early stage and being maintained through growth of the epithelial structures. Under suspension culture with COL, the polarization was impaired with formation of multi-lumens on the tubes, implying the importance of ECM microenvironment for tubulogenesis. Our results suggest a mechanism for cells to form polarity and coordinate positioning in vivo, and a strategy for engineering epithelial structures through cell-soluble ECM interaction and self-assembly in vitro.

scholarly journals Sox8 is essential for M cell maturation to accelerate IgA response at the early stage after weaning in mice

2019 ◽  
Vol 216 (4) ◽  
pp. 831-846 ◽  
Author(s):  
Shunsuke Kimura ◽  
Nobuhide Kobayashi ◽  
Yutaka Nakamura ◽  
Takashi Kanaya ◽  
Daisuke Takahashi ◽  
...  
Keyword(s):  
Immune Responses ◽  
Early Stage ◽  
Biological Significance ◽  
M Cells ◽  
M Cell ◽  
Antigen Uptake ◽  
Molecular Machinery ◽  
Iga Response ◽  
Genetic Deletion ◽  
Mucosal Immune Responses

Microfold (M) cells residing in the follicle-associated epithelium (FAE) of the gut-associated lymphoid tissue are specialized for antigen uptake to initiate mucosal immune responses. The molecular machinery and biological significance of M cell differentiation, however, remain to be fully elucidated. Here, we demonstrate that Sox8, a member of the SRY-related HMG box transcription factor family, is specifically expressed by M cells in the intestinal epithelium. The expression of Sox8 requires activation of RANKL-RelB signaling. Chromatin immunoprecipitation and luciferase assays revealed that Sox8 directly binds the promoter region of Gp2 to increase Gp2 expression, which is the hallmark of functionally mature M cells. Furthermore, genetic deletion of Sox8 causes a marked decrease in the number of mature M cells, resulting in reduced antigen uptake in Peyer’s patches. Consequently, juvenile Sox8-deficient mice showed attenuated germinal center reactions and antigen-specific IgA responses. These findings indicate that Sox8 plays an essential role in the development of M cells to establish mucosal immune responses.

scholarly journals In VivoStudy of Ligament-Bone Healing after Anterior Cruciate Ligament Reconstruction Using Autologous Tendons with Mesenchymal Stem Cells Affinity Peptide Conjugated Electrospun Nanofibrous Scaffold

2013 ◽  
Vol 2013 ◽  
pp. 1-11 ◽  
Author(s):  
Jingxian Zhu ◽  
Xin Zhang ◽  
Zhenxing Shao ◽  
Linghui Dai ◽  
La Li ◽  
...  
Keyword(s):  
Acl Reconstruction ◽  
Bone Healing ◽  
Early Stage ◽  
Cruciate Ligament ◽  
Nanofibrous Scaffold ◽  
Control Group ◽  
Type I ◽  
Specific Staining ◽  
Affinity Peptide

Electrospinning nanofibrous scaffold was commonly used in tissue regeneration recently. Nanofibers with specific topological characteristics were reported to be able to induce osteogenic differentiation of MSCs. In thisin vivostudy, autologous tendon grafts with lattice-like nanofibrous scaffold wrapping at two ends of autologous tendon were used to promote early stage of ligament-bone healing after rabbit ACL reconstruction. To utilize native MSCs from bone marrow, an MSCs specific affinity peptide E7 was conjugated to nanofibrous meshes. After 3 months, H-E assessment and specific staining of collagen type I, II, and III showed direct ligament-bone insertion with typical four zones (bone, calcified fibrocartilage, fibrocartilage, and ligament) in bioactive scaffold reconstruction group. Diameters of bone tunnel were smaller in nanofibrous scaffold conjugated E7 peptide group than those in control group. The failure load of substitution complex also indicated a stronger ligament-bone insertion healing using bioactive scaffold. In conclusion, lattice-like nanofibrous scaffold with specific MSCs affinity peptide has great potential in promoting early stage of ligament-bone healing after ACL reconstruction.

scholarly journals Differential expression of lectin-binding sites defines mouse intestinal M-cells.

1993 ◽  
Vol 41 (11) ◽  
pp. 1679-1687 ◽  
Author(s):  
M A Clark ◽  
M A Jepson ◽  
N L Simmons ◽  
T A Booth ◽  
B H Hirst
Keyword(s):  
Electron Microscope ◽  
Laser Scanning ◽  
Lectin Binding ◽  
Fluorescein Isothiocyanate ◽  
Goblet Cells ◽  
Confocal Laser ◽  
M Cells ◽  
M Cell ◽  
Ulex Europaeus ◽  
Transmission Electron

We investigated the binding of four lectins to the follicle-associated epithelium (FAE) overlying fixed mouse small intestinal Peyer's patches to identify M-cell-specific surface markers. Wheat germ agglutinin and peanut agglutinin displayed heterogeneous staining patterns, binding most avidly to the intestine goblet cells. In contrast, the lectins Ulex europaeus 1 (UEA 1) and Psophocarpus tetragonolobus (winged bean; WBA) were almost exclusively M-cell specific. When confocal laser scanning images of tissues stained with fluorescein isothiocyanate (FITC)-conjugated UEA1 or WBA were compared with the appearance of the same tissues under the scanning electron microscope (SEM), UEA1 strongly stained 97.2% (106/109) of M-cells, 0.6% (3/516) enterocytes, and 0% (0/28) goblet cells, whereas WBA stained 100% (83/83) M-cells, 1.7% (6/361) enterocytes, and 5.3% (1/19) goblet cells. The M-cell specificity of the lectin binding was further demonstrated by localization of horseradish peroxidase (HRP)-conjugated lectins under the transmission electron microscope (TEM). This is the first demonstration of carbohydrates in the glycocalyx of M-cells that are not expressed elsewhere on the FAE surface. These carbohydrates not only provide a means to identify mouse M-cells by LM but may also contribute to the occurrence of specific interactions between microorganisms and the M-cell apical membrane.

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