No abstract

SummaryYersinia enterocolitica cross the intestinal epithelium via translocation through M cells, which are located in the follicle-associated epithelium (FAE) of Peyer's patches (PP). To investigate the molecular basis of this process, studies were performed using a recently developed in vitro model, in which the enterocyte-like cell line Caco-2 and PP lymphocytes are co-cultured in order to establish FAE-like structures including M cells. Here, we demonstrate that Y. enterocolitica does not adhere significantly to the apical membrane of differentiated enterocyte-like Caco-2 cells that express binding sites for Ulex europaeus agglutinin (UEA)-1. In contrast, Y. enterocolitica adhered to, and was internalized by, cells that lacked UEA-1 binding sites and displayed a disorganized brush border. These cells were considered to be converted to M-like cells. Further analysis revealed that part of these cells expressed b1 integrins at their apical surface and, as revealed by comparison of wild-type and mutant strains, interacted with invasin of Y. enterocolitica. Consistently, anti-b1 integrin antibodies significantly inhibited internalization of inv-expressing yersiniae. Experiments with Yersinia mutant strains deficient in YadA or Yop secretion revealed that these virulence factors play a minor role in this process. After internalization, yersiniae were transported within LAMP-1-negative vacuoles to, and released at, the basal surface. Internalization and transport of yersiniae was inhibited by cytochalasin D, suggesting that F-actin assembly is required for this process. These results provide direct evidence that expression of b1 integrins at the apical surface of M cells enables interaction with the invasin of Y. enterocolitica, and thereby initiates internalization and translocation of bacteria.

Material Supplementary 2.DC1http://www.jimmunol.org/content/suppl/2010/06/04/jimmunol.090391

SUMMARY Lampreys, one of the only two surviving groups of agnathan (jawless)vertebrates, contain several anadromous species that, during their life cycle,thus migrate from fresh to seawater and back to freshwater. Lampreys have independently evolved the same overall osmoregulatory mechanisms as the gnathostomatous (jawed) and distantly related teleost fishes. Lamprey gills thus likewise play a central role in taking up and secreting monovalent ions. However, the ultrastructural characteristics and distribution of their epithelial cell types [ammocoete mitochondria-rich (MR) cell, intercalated MR cell, chloride cell and pavement cell] differ in several respects from those of teleosts. The ultrastructural characteristics of these cells are distinctive and closely resemble those of certain ion-transporting epithelia in other vertebrates, for which the function has been determined. The data on each cell type, together with the stage in the life cycle at which it is found, i.e. whether in fresh or seawater, enable the following proposals to be made regarding the ways in which lampreys use their gill epithelial cells for osmoregulating in hypo- and hypertonic environments. In freshwater, the intercalated MR cell takes up Cl– and secretes H+,thereby facilitating the uptake of Na+ through pavement cells. In seawater, the chloride cell uses a secondarily active transcellular transport of Cl– to provide the driving force for the passive movement of Na+ through leaky paracellular pathways between these cells.

The occurrence of cytokeratins, vimentin, and desmin in the dome epithelia and adjacent non-dome epithelia in four locations of gut-associated lymphoid tissues (GALT) of adult and newborn rabbits (Peyer's patches, sacculus rotundus, caecal lymphoid patches and appendix) was studied with monoclonal antibodies, using the indirect immunoperoxidase technique. In all locations investigated in adult animals, antibodies specific for vimentin labelled (1) M-cells, which engulf intra-epithelial lymphocytes, (2) columnar epithelial cells at the base of the domes lacking an apparent contact with lymphocytes ("immature" M-cells), and (3) flat cells, which lie in the lamina propria under the dome epithelium, and which line the basal lamina with thin cytoplasmic processes. In newborn rabbits, columnar epithelial cells resembling the immature M-cells of adults were selectively stained with vimentin antibodies. In M-cells, the strongest immunoreactivity was present in the perinuclear region and close to the pocket membrane, whereas the most apical and most basal parts of the cytoplasm showed no vimentin-immunoreactivity. Enterocytes in the dome epithelium and in the non-dome epithelium were vimentin-negative. M-cells and enterocytes bound antibodies against cytokeratin peptides 18 and 19 in adult and newborn animals. Compared with enterocytes, M-cells showed less intense staining for cytokeratins. Dome epithelia and no-dome epithelia did not contain desmin-immunoreactive cells. The results suggest that vimentin is a sensitive marker for M-cells in rabbit GALT.