Background:; We have previously demonstrated that pulmonary collectins (SP-A and SP-D) attenuate intracellular growth of Legionella pneumophila. However, the molecular mechanisms have not been completely understood. To clarify the molecular mechanisms involved in inhibitory effect of pulmonary collectins. Methods: Using adenylate cyclase-reporter assay system, we quantitatively evaluated the activity of Type IV secretion system (TIVSS). Furthermore, we examined autophagic activity in THP-1 cells infected with L. pneumophila. Induction of autophagy was evaluated by western blotting using anti-LC3 and anti-p62 antibodies. Results: Pulmonary collectins inhibited secretion of effector molecules through TIVSS. Interaction between collectins and L. pneumophila seemed to be responsible for inhibitory effect. It has been reported that effector molecules secreted from L. pneumophila cause the delay in induction of autophagy. Consistent with the results in above, pulmonary collectins accelerated the induction of autophagy in L. pneumophila-infected cells. Conclusions: Delay of autophagy is crucial for intracellular growth of L. pneumophila. Our data suggest that pulmonary collectins attenuate the activity of TIVSS, and thereby accelerate induction of autophagy. Results in the present study clarify one of the molecular mechanisms by which pulmonary collectins inhibit intracellular growth of L. pneumophila.
The imager that observes the ground surface with high spatial resolution from geostationary orbit has been studied. To obtain image data with high ground sampling distance (GSD) from a geostationary orbit, an extremely large-diameter and long-focal-length telescope is required. In order to realize this imager, we plan to adopt a synthetic aperture type reflection optical telescope (Korsch type) in which the primary mirror is composed of six segmented mirrors. The diameter of the synthesized aperture telescope is approximately 3.6m, and the focal length is approximately 30 m. Prior to the development of the actual imager, we have been producing a full-scale prototype of the segmented mirror that constitutes the primary mirror and its supporting structure. The aperture shape of the prototype segmented mirror is hexagonal, and its diagonal length is approximately 1.35 m. The supporting structure of the prototype segmented mirror incorporates a mechanism (6-axes adjustment mechanism) for adjusting the alignment of the segmented mirror with six degrees of freedom. A mechanism for adjusting the curvature of the segmented mirror (curvature adjustment mechanism) was also incorporated. In this paper, the design, the manufacturing, and the testing status of this prototype are described.
Bacterial lipopolysaccharide (LPS)-induced exocytosis of granular hemocytes is a key component of the horseshoe crab's innate immunity to infectious microorganisms; stimulation by LPS induces the secretion of various defense molecules from the granular hemocytes. Using a previously uncharacterized assay for exocytosis, we clearly show that hemocytes respond only to LPS and not to other pathogen-associated molecular patterns, such as β-1,3-glucans and peptidoglycans. Furthermore, we show that a granular protein called factor C, an LPS-recognizing serine protease zymogen that initiates the hemolymph coagulation cascade, also exists on the hemocyte surface as a biosensor for LPS. Our data demonstrate that the proteolytic activity of factor C is both necessary and sufficient to trigger exocytosis through a heterotrimeric GTP-binding protein-mediating signaling pathway. Exocytosis of hemocytes was not induced by thrombin, but it was induced by hexapeptides corresponding to the tethered ligands of protease-activated G protein-coupled receptors (PARs). This finding suggested the presence of a PAR-like receptor on the hemocyte surface. We conclude that the serine protease zymogen on the hemocyte surface functions as a pattern-recognition protein for LPS.
The adherence of uropathogenic Escherichia coli (UPEC) to the host urothelial surface is the first step for establishing UPEC infection. Uroplakin Ia (UPIa), a glycoprotein expressed on bladder urothelium, serves as a receptor for FimH, a lectin located at bacterial pili, and their interaction initiates UPEC infection. Surfactant protein D (SP-D) is known to be expressed on mucosal surfaces in various tissues besides the lung. However, the functions of SP-D in the non-pulmonary tissues are poorly understood. The purposes of this study were to investigate the possible function of SP-D expressed in the bladder urothelium and the mechanisms by which SP-D functions. SP-D was expressed in human bladder mucosa, and its mRNA was increased in the bladder of the UPEC infection model in mice. SP-D directly bound to UPEC and strongly agglutinated them in a Ca2+-dependent manner. Co-incubation of SP-D with UPEC decreased the bacterial adherence to 5637 cells, the human bladder cell line, and the UPEC-induced cytotoxicity. In addition, preincubation of SP-D with 5637 cells resulted in the decreased adherence of UPEC to the cells and in a reduced number of cells injured by UPEC. SP-D directly bound to UPIa and competed with FimH for UPIa binding. Consistent with the in vitro data, the exogenous administration of SP-D inhibited UPEC adherence to the bladder and dampened UPEC-induced inflammation in mice. These results support the conclusion that SP-D can protect the bladder urothelium against UPEC infection and suggest a possible function of SP-D in urinary tract. The adherence of uropathogenic Escherichia coli (UPEC) to the host urothelial surface is the first step for establishing UPEC infection. Uroplakin Ia (UPIa), a glycoprotein expressed on bladder urothelium, serves as a receptor for FimH, a lectin located at bacterial pili, and their interaction initiates UPEC infection. Surfactant protein D (SP-D) is known to be expressed on mucosal surfaces in various tissues besides the lung. However, the functions of SP-D in the non-pulmonary tissues are poorly understood. The purposes of this study were to investigate the possible function of SP-D expressed in the bladder urothelium and the mechanisms by which SP-D functions. SP-D was expressed in human bladder mucosa, and its mRNA was increased in the bladder of the UPEC infection model in mice. SP-D directly bound to UPEC and strongly agglutinated them in a Ca2+-dependent manner. Co-incubation of SP-D with UPEC decreased the bacterial adherence to 5637 cells, the human bladder cell line, and the UPEC-induced cytotoxicity. In addition, preincubation of SP-D with 5637 cells resulted in the decreased adherence of UPEC to the cells and in a reduced number of cells injured by UPEC. SP-D directly bound to UPIa and competed with FimH for UPIa binding. Consistent with the in vitro data, the exogenous administration of SP-D inhibited UPEC adherence to the bladder and dampened UPEC-induced inflammation in mice. These results support the conclusion that SP-D can protect the bladder urothelium against UPEC infection and suggest a possible function of SP-D in urinary tract.
