Mami Mutoh

RANKL regulates differentiation of microfold cells in mouse nasopharynx-associated lymphoid tissue (NALT)

Murine nasopharynx-associated lymphoid tissue (NALT), located at the base of the nasal cavity, serves as a major site for the induction of mucosal immune responses against airway antigens. The follicle-associated epithelium (FAE) covering the luminal surface of NALT is characterized by the presence of microfold cells (M cells), which take up and transport luminal antigens to lymphocytes. Glycoprotein 2 (GP2) has recently been identified as a reliable marker for M cells in Peyer’s patches of the intestine. However, the expression of GP2 and other functional molecules in the M cells of NALT has not yet been examined. We have immunohistochemically detected GP2-expressing cells in the FAE of NALT and the simultaneous expression of other intestinal M-cell markers, namely Tnfaip2, CCL9, and Spi-B. These cells have been further identified as M cells because of their higher uptake capacity of luminal microbeads. Electron microscopic observations have shown that GP2-expressing cells on the FAE display morphological features typical of M cells: they possess short microvilli and microfolds on the luminal surface and are closely associated with intraepithelial lymphocytes. We have also found that the receptor activator of nuclear factor kappa-B ligand (RANKL) is expressed by stromal cells underneath the FAE, which provides its receptor RANK. The administration of RANKL markedly increases the number of GP2+Tnfaip2+ cells on the NALT FAE and that of intestinal M cells. These results suggest that GP2+Tnfaip2+ cells in NALT are equivalent to intestinal M cells, and that RANKL-RANK signaling induces their differentiation.

GP2-expressing cells in the conjunctiva and tear ducts of mice: identification of a novel type of cells in the squamous stratified epithelium.

GP2 is a membrane-associated secretory protein originally identified in zymogen granules of pancreatic acinar cells. Recently, this glycoprotein has attracted attention as a marker substance of M cells of Peyers patches and for its involvement in the selective uptake of pathological bacteria via M cells. When we stained the conjunctiva and tear ducts of mice using a GP2 antibody, all goblet cells in the squamous stratified epithelium of the conjunctiva were intensely immunolabeled, while goblet cells in the intestine and airway were devoid of the immunoreactivity, indicating that the conjunctiva contains a special type of goblet cell. Further immunostaining for GP-2 labeled dispersed cells of peculiar shapes within the stratified squamous epithelium in the lacrimal canaliculi, lacrimal sac, and nasolacrimal duct. The GP2-immunoreactive cells in the tear duct projected arched or branched processes toward the basement membrane. Electron-microscopically, immunogold particles for GP2 outlined the basolateral plasma membrane of both the conjuntival goblet cells and the peculiarly shaped cells in the tear duct. Intracellularly, GP2 products of the goblet cells were localized around secretory granules in the apical cytoplasm and those of the tear duct cells inside the vesicles. The luminal contents close to apical plasma membrane were heavily labeled with immunogold particles, suggesting an exocytosis-based targeting of GP2 to the plasma membrane and its release into the lumen. The possible function of GP2 in tear ducts is discussed in relation to a defense system against invasive microoranisms and antigens.

Internal distribution of micro- / nano-sized inorganic particles and their cytocompatibility

Nano-sized materials have received much attention lately, both in terms of their multiple applications and their biocompatibility. From both viewpoints, understanding the biodistribution of administered nano-materials is very important. In this study, we succeeded in visualizing the biodistribution of administered nano-materials using a scanning X-ray analytical microscope and magnetic resonance imaging method. Quantitative observation was carried out by inductively coupled plasma – atomic emission spectroscopy. We observed that the administered nano-particles accumulated in the liver, lung and spleen of mice. To estimate their cytocompatibility, the nano-particles were exposed to human liver cells. The results suggested that the micro-/ nano- particles have good cytocompatibility, except for copper oxide nano-particles.

Density functional theory study on oligosilane-functionalized C60 fullerene

Oligosilane-functionalized C60 fullerenes [namely, C60–(SiH2) n –H, n = 1–4] have been investigated by the density functional theory (DFT) method to elucidate the structures and electronic states of oligosilane-radical added fullerene. The DFT calculation showed that oligosilane radicals bind to the carbon atom of C60 in the on-top site, and a strong Si–C heterojunction is formed. The binding energies of oligosilane radicals to C60 were calculated to be 24.6–28.2 kcal/mol at the CAM-B3LYP/6-311G(d,p) level. The electronic states of oligosilane-functionalized fullerenes C60–(SiH2) n –H are discussed on the basis of theoretical results.

Cytocompatiblity of Ceramic Nanoparticles to Various Types of Cells

In this study, we investigated the cytocompatibility of ceramic nanoparticles on different types of cells. All ceramics nanoparticles investigated in this study except Copper oxide (CuO) exhibited good cytocompatibility and cell viability (90% or more) even at 20 ppm concentration. In contrast, CuO nanoparticles caused cell inflammation, and their effect depended on their particle size. Confocal fluorescence microscopy measurements indicated that some particles had penetrated into the cells. These results indicate that except CuO nanoparticles, all other ceramic nanoparticles reported herein exhibited excellent cytocompatibility even for lung epithelial cells.