Recently, autophagy has been associated with the TLR signaling pathway to eliminate intracellular pathogens in innate immune system. However, it is unknown if other pathways regulate during immunologic response. Given critical role of purinergic P2X7 receptor (P2X7R) various functions (i.e., caspase activation and IL-1beta secretion), principal objective here was determine whether P2X7R may cells. We observed both MG6 mouse microglial cells primary microglia that by ATP increases expression...

Prion diseases are fatal neurodegenerative disorders and no effective treatment has been established to date. In this study, we evaluated the effect of FK506 (tacrolimus), a macrolide that is known be mild immunosuppressant, on prion infection, using cell culture animal models. We found markedly reduced abnormal form protein (PRNPSc) in cultures (N2a58 MG20) infected with Fukuoka-1 prion. The levels autophagy-related molecules such as LC3-II, ATG12–ATG5 ATG7 were significantly increased...

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a key glycolytic enzyme that predominantly localized in the cytoplasm. However, recent studies have suggested GAPDH released by various cells and extracellular involved regulation of neuritogenesis neuronal cells. It has also been reported expressed on surfaces macrophages functions as transferrin receptor. since leaderless protein mechanisms which it reaches environment remain unclear. Here, we examined role P2X7 receptor (P2X7R), an...

Plant polyphenols, RG-tannin, and applephenon had been reported to inhibit cholera toxin (CT) ADP-ribosyltransferase activity CT-induced fluid accumulation in mouse ileal loops. A high molecular weight fraction of hop bract extract (HBT) also inhibited CT activity. We report here the effect those polyphenols on binding entry into Vero cells. Binding cells or ganglioside GM1, a receptor, was concentration-dependent manner by HBT but not RG-tannin. These observations were confirmed...

Background Prions, infectious agents associated with prion diseases such as Creutzfeldt-Jakob disease in humans, bovine spongiform encephalopathy (BSE) cattle, and scrapie sheep goats, are primarily comprised of PrPSc, a protease-resistant misfolded isoform the cellular protein PrPC. Protein misfolding cyclic amplification (PMCA) is highly sensitive technique used to detect minute amounts PrPSc. However, current PMCA has been unsuccessful achieving good cattle. The detailed distribution...

mAbs T1 and T2 were established by immunizing PrP gene ablated mice with recombinant MoPrP of residues 121-231. Both cross-reactive from hamster, sheep, cattle deer. A linear epitope mAb was identified at 137-143 buried in PrP(C) expressed on the cell surface. showed no inhibitory effect accumulation PrP(Sc) cultured scrapie-infected neuroblastoma (ScN2a) cells. In contrast, recognized a discontinuous ranged on, or structured by, 132-217 this exposed surface PrP(C). an excellent vitro 50%...

Abstract The P2X7 receptor (P2X7R), an ATP-gated ion channel, plays essential roles in the release and maturation of IL-1β microglial cells brain. Previously, we found that lysophosphatidylcholine (LPC) potentiated P2X7R-mediated intracellular signals cells. In this study, determined whether lysophospholipids, i.e., LPC sphingosylphosphorylcholine (SPC), modulate ATP-induced processing mediated by P2X7R mouse MG6 or SPC alone induced precursor (pro-IL-1β) mature (mIL-1β) from LPS-primed...

It has been assumed that the agent causing BSE in cattle is a uniform strain (classical BSE); however, different neuropathological and molecular phenotypes of (atypical BSE) have recently reported. We demonstrated successful transmission L-type-like atypical detected Japan (BSE/JP24 isolate) to cattle. Based on incubation period, hallmarks, properties abnormal host prion protein, characteristics BSE/JP24 were apparently distinguishable from classical closely resemble those bovine amyloidotic...

Atypical bovine spongiform encephalopathy (BSE) has recently been identified in Europe, North America, and Japan. It is classified as H-type L-type BSE according to the molecular mass of disease-associated prion protein (PrPSc). To investigate topographical distribution deposition patterns immunolabeled PrPSc, isolate was inoculated intracerebrally into cattle. successfully transmitted 3 calves, with incubation periods between 500 600 days. Moderate severe changes were detected cerebral...

Surveillance for bovine spongiform encephalopathy (BSE) in fallen stock Japan is conducted with a commercial enzyme-linked immunosorbent assay (ELISA) mass screening, Western blotting (WB) and immunohistochemistry performed confirmation of the ELISA. All tests are based on immunological detection an abnormal isoform prion protein (PrPSc) brain tissues, which have sometimes deteriorated by time samples from reach diagnostic laboratory. To evaluate BSE surveillance procedures stock, we...

ABSTRACT Eight hundred Erysipelothrix strains isolated between 1992 and 2002 from swine with erysipelas in Japan were serotyped. Thirty-seven, 47, 73, 643 animals acute septicemia, urticaria, chronic endocarditis, arthritis, respectively, of which 381, 146, 254, 19 isolates belonged to serotypes 1a, 1b, 2b other serotypes, respectively. All serotype 1a further examined for acriflavine resistance their genotypes discriminate them the attenuated live vaccine strain, defined as is resistant...

Several lines of evidence suggest that microglia have important roles in the pathogenesis prion diseases. Here, we establish a novel microglial cell line (MG20) from neonatal tga20 mice overexpress murine protein. After exposure to Chandler scrapie, observed replication and accumulation disease-associated forms protein MG20 cells up 15th passage. Furthermore, were susceptible ME7, Obihiro bovine spongiform encephalopathy agents. Thus, persistently infected with various strains provide useful...

A case of L-type-like atypical bovine spongiform encephalopathy was detected in 14-year-old Japanese black beef cattle (BSE/JP24). To clarify the biological and biochemical properties prion BSE/JP24, we performed a transmission study with wild-type mice bovinized transgenic (TgBoPrP). The BSE/JP24 transmitted to TgBoPrP incubation period 199.7 ± 3.4 days, which shorter than that classical BSE (C-BSE) (223.5 13.5 days). Further, C-BSE about 409 whereas inoculated showed no clinical signs up...

The potent pro-inflammatory cytokine, interleukin-1β (IL-1β), is synthesized as an inactive 33-kDa precursor (pro-IL-1β) and processed by caspase 1 into the bioactive 17-kDa mature form. P2X7 receptor, ATP-gated cation channel, plays essential role in activation, production release of We recently reported ATP induces unconventional 20-kDa form IL-1β (p20-IL-1β) from lipopolysaccharide-primed microglial cells. Emerging evidence suggests physiological relevance for p20-IL-1β; however,...

Background Prion diseases are fatal neurodegenerative disorders with no effective therapy currently available. Accumulating evidence has implicated over-activation of P2X7 ionotropic purinergic receptor (P2X7R) in the progression neuronal loss several diseases. This led to speculation that simultaneous blockade this and prion replication can be an therapeutic strategy for We have focused on Brilliant Blue G (BBG), a well-known P2X7R antagonist, possessing chemical structure expected confer...

The precise mechanism underlying the conversion of normal prion protein (PrPC) into abnormal (PrPSc) remains unclear. Protein misfolding cyclic amplification (PMCA), an in vitro technique used for amplifying PrPSc, results PrPSc replication that preserves strain-specific characteristics input PrPSc; thus, PMCA mimics process vivo replication. Previous work has demonstrated PMCA, nucleic acids are critical amplification, but little information been reported on glycosaminoglycan (GAG)...

Abstract Prion diseases are fatal neurodegenerative disorders, and the conformational conversion of normal cellular prion protein (PrP C ) into its pathogenic, amyloidogenic isoform Sc is essential event in pathogenesis these diseases. Lactoferrin (LF) a cationic iron‐binding glycoprotein belonging to transferrin (TF) family, which accumulates amyloid deposits brain such as Alzheimer’s disease Pick’s disease. In present study, we have examined effects LF on PrP formation by using cell...

Background Sheep scrapie is caused by multiple prion strains, which have been classified on the basis of their biological characteristics in inbred mice. The heterogeneity natural prions individual sheep and flocks has not clearly defined. Methodology/Principal Findings In this study, we intravenously injected 2 (Suffolk Corriedale) with material from a case breed). These 3 had identical protein (PrP) genotypes. protease-resistant core PrP (PrPres) experimental Suffolk was similar to that...

We recently established mouse microglial cells persistently infected with mouse‐adapted scrapie ME7 (ScMG20/ME7) for in vitro study of prion pathogenesis. Here, we found that ScMG20/ME7 were hypersensitive to P2X7 receptor agonists, as demonstrated by sustained Ca 2+ influx, membrane pore formation, cell death, and interleukin‐1β release. mRNA expression was upregulated these cells, also scrapie‐infected mice brains. Treatment pentosan polysulfate eliminated the infectivity disease‐related...