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Langerhans cells (LCs) are mainly present in the epidermis and mucosa, and have important roles during skin infection. Migration of LCs to lymph nodes is essential for antigen presentation. However, due to the difficulties in isolating and culturing human LCs, it is not fully understood how LCs move and interact with the extracellular matrix (ECM) through their adhesion molecules such as integrin, during the immune responses. In this study, we aimed to investigate LC motility, cell shape and the role of integrin under inflammatory conditions using monocyte‐derived Langerhans cells (moLCs) as a model. As a result, lipopolysaccharide (LPS) stimulation increased adhesion on fibronectin coated substrate and integrin α5 expression in moLCs. Time‐lapse imaging of moLCs revealed that stimulation with LPS elongated cell shape, whilst decreasing their motility. Additionally, this decrease in motility was not observed when pre‐treated with a neutralising antibody targeting integrin α5. Together, our data suggested that activation of LCs decreases their motility by promoting integrin α5 expression to enhance their affinity to the fibronectin, which may contribute to their migration during inflammation. [ABSTRACT FROM AUTHOR]

This article discusses a rare case of co-occurrence of bullous pemphigoid (BP) and mucosal-dominant pemphigus vulgaris (MDPV) in a Japanese man. The patient presented with tense blisters and erosions on the skin and mucosal lesions on the lips, tongue, and buccal mucosa. Initially, the mucosal lesions were attributed to BP, but further analysis revealed that they were caused by pathogenic anti-desmoglein 3 (Dsg3) antibodies. The article emphasizes the importance of confirming the pathogenicity of anti-Dsg antibodies and highlights the usefulness of a bead aggregation assay in monitoring disease activity and selecting appropriate treatments. [Extracted from the article]

Bacterial infections can activate and mobilize hematopoietic stem and progenitor cells (HSPCs) from the bone marrow (BM) to the spleen, a process termed extramedullary hematopoiesis (EMH). Recent studies suggest that commensal bacteria regulate not only the host immune system but also hematopoietic homeostasis. However, the impact of gut microbes on hematopoietic pathology remains unclear. Here, we find that systemic single injections of Akkermansia muciniphila (A. m.), a mucin‐degrading bacterium, rapidly activate BM myelopoiesis and slow but long‐lasting hepato‐splenomegaly, characterized by the expansion and differentiation of functional HSPCs, which we term delayed EMH. Mechanistically, delayed EMH triggered by A. m. is mediated entirely by the MYD88/TRIF innate immune signaling pathway, which persistently stimulates splenic myeloid cells to secrete interleukin (IL)‐1α, and in turn, activates IL‐1 receptor (IL‐1R)‐expressing splenic HSPCs. Genetic deletion of Toll‐like receptor‐2 and ‐4 (TLR2/4) or IL‐1α partially diminishes A. m.‐induced delayed EMH, while inhibition of both pathways alleviates splenomegaly and EMH. Our results demonstrate that cooperative IL‐1R‐ and TLR‐mediated signals regulate commensal bacteria‐driven EMH, which might be relevant for certain autoimmune disorders. Synopsis: A single injection of Akkermansia muciniphila, one of the major mucin‐degrading microbial species in the human gut, induces long‐lasting hepatosplenomegaly with splenic extramedullary hematopoiesis (EMH) by activating cooperative IL‐1R‐ and TLR‐mediated innate immune pathways.Injection of A. muciniphila lysate induces long‐lasting anemia and hepatosplenomegaly with expansion of functional HSCs.Cell expansion in the spleen is mediated by partially TLR2/4‐dependent and entirely MYD88/TRIF‐dependent pathways.Sustained secretion of IL‐1α from splenic myeloid cells contributes to TLR2/4‐independent EMH.Pharmacological inhibition of IL‐1R in addition to genetic ablation of TLR2/4 can completely alleviate induced EMH. [ABSTRACT FROM AUTHOR]

Mitochondrial DNA (mtDNA) leakage into the cytoplasm can occur when cells are exposed to noxious stimuli. Specific sensors recognize cytoplasmic mtDNA to promote cytokine production. Cytoplasmic mtDNA can also be secreted extracellularly, leading to sterile inflammation. However, the mode of secretion of mtDNA out of cells upon noxious stimuli and its relevance to human disease remain unclear. Here, we show that pyroptotic cells secrete mtDNA encapsulated within exosomes. Activation of caspase‐1 leads to mtDNA leakage from the mitochondria into the cytoplasm via gasdermin‐D. Caspase‐1 also induces intraluminal membrane vesicle formation, allowing for cellular mtDNA to be taken up and secreted as exosomes. Encapsulation of mtDNA within exosomes promotes a strong inflammatory response that is ameliorated upon exosome biosynthesis inhibition in vivo. We further show that monocytes derived from patients with Behçet's syndrome (BS), a chronic systemic inflammatory disorder, show enhanced caspase‐1 activation, leading to exosome‐mediated mtDNA secretion and similar inflammation pathology as seen in BS patients. Collectively, our findings support that mtDNA‐containing exosomes promote inflammation, providing new insights into the propagation and exacerbation of inflammation in human inflammatory diseases. Synopsis: Mitochondrial DNA (mtDNA) secreted by pyroptotic cells is encapsulated within exosomes and induces excessive inflammation resembling symptoms seen in patients with Behçet's syndrome. In pyroptotic cells, the activation of caspase‐1 and subsequent action of gasdermin‐D promote the efflux of mtDNA from the mitochondria into the cytoplasm.Activation of caspase‐1 promotes the formation of intracellular vesicles (ILVs), mtDNA uptake into ILVs, and its release into the extracellular space via exosomes.The encapsulation of mtDNA within exosomes promotes inflammation, leukocyte mobilization, and cytokine production.Patients with Behçet's syndrome show increased mtDNA in the blood and caspase‐1‐mediated mitochondrial damage in the isolated monocytes.In patients with Behçet's syndrome monocytes secrete mtDNA‐containing exosomes that induce pathological inflammation in an NLRP3‐ and TLR9‐dependent manner. [ABSTRACT FROM AUTHOR]

Primary cilia influence cell activity, and thus have a unique role in maintaining cell proliferation and differentiation. In atopic dermatitis (AD) and psoriasis, areas of skin inflammation exhibit dysregulated keratinocyte homeostasis. The role of primary cilia in these conditions remains unclear. The objectives of this study is to elucidate the incidence of primary cilia in skin inflammation and the potential mechanism underlying the dysregulation of keratinocytes. Primary cilia were observed using immunofluorescence staining. Normal skin samples were compared with skin samples from patients with AD or psoriasis in terms of cilia numbers and length. The effect of cytokine stimulation on ciliogenesis in keratinocytes was analysed using a primary keratinocyte culture. IFT88, an important ciliary intraflagellar protein, was blocked in Th2 and Th17 cytokines‐stimulated keratinocytes. These effects were analysed with quantitative polymerase chain reaction and Western blot. Significant increases in ciliated cells were observed in AD and psoriasis skin samples compared with normal skin samples. The stimulation of keratinocytes using Th2 and Th17 cytokines modulated the formation of primary cilia. The amount of IFT88 in the primary cilia associated with the phosphorylation of JNK, but not p38, in keratinocytes stimulated with interleukin‐13, 17A and 22. An increase of ciliated cells in the epidermis may impair keratinocyte differentiation under stress conditions caused by inflammation in both AD and psoriasis patients. [ABSTRACT FROM AUTHOR]

Immune potentiators, termed adjuvants, trigger early innate immune responses to ensure the generation of robust and long‐lasting adaptive immune responses of vaccines. Presented here is a study that takes advantage of a self‐assembling small‐molecule library for the development of a novel vaccine adjuvant. Cell‐based screening of the library and subsequent structural optimization led to the discovery of a simple, chemically tractable deoxycholate derivative (molecule 6, also named cholicamide) whose well‐defined nanoassembly potently elicits innate immune responses in macrophages and dendritic cells. Functional and mechanistic analyses indicate that the virus‐like assembly enters the cells and stimulates the innate immune response through Toll‐like receptor 7 (TLR7), an endosomal TLR that detects single‐stranded viral RNA. As an influenza vaccine adjuvant in mice, molecule 6 was as potent as Alum, a clinically used adjuvant. The studies described here pave the way for a new approach to discovering and designing self‐assembling small‐molecule adjuvants against pathogens, including emerging viruses. [ABSTRACT FROM AUTHOR]

Background and Aim: The usefulness of preventive closure of the frenulum after endoscopic papillectomy (EP) could reduce bleeding. The feasibility and safety of clipping were evaluated in this prospective pilot study. Methods: This study involved 40 consecutive patients who underwent preventive closure of the frenulum by clipping just after EP. The outcome data were compared with those of the previous 40 patients in whom no preemptive closure had been performed (no‐closure group) (UMIN000014783). Additionally, the bleeding sites were examined. Results: The clipping procedure was successful in all patients. As compared to the no‐closure group, the rate of bleeding (P = 0.026) and period of hospital stay (P < 0.001) were significantly reduced in the closure group. There was no difference in the procedure time between the two groups. Furthermore, the incidence rates of pancreatitis and perforation were comparable in the two groups. The bleeding was noted in the frenulum area rather than at any other site in 90.9% of cases. Conclusion: Preventive closure of the frenulum after EP is an effective, safe, rational, and economical method to reduce the incidence of delayed bleeding, without prolonging the procedure time or increasing the risk of post‐procedure pancreatitis perforation. [ABSTRACT FROM AUTHOR]

Adjuvants improve the potency of vaccines, but the modes of action (MOAs) of most adjuvants are largely unknown. TLR‐dependent and ‐independent innate immune signaling through the adaptor molecule MyD88 has been shown to be pivotal to the effects of most adjuvants; however, MyD88's involvement in the TLR‐independent MOAs of adjuvants is poorly understood. Here, using the T‐dependent antigen NIPOVA and a unique particulate adjuvant called synthetic hemozoin (sHZ), we show that MyD88 is required for early GC formation and enhanced antibody class‐switch recombination (CSR) in mice. Using cell‐type‐specific MyD88 KO mice, we found that IgG2c class switching, but not IgG1 class switching, was controlled by B cell‐intrinsic MyD88 signaling. Notably, IFN‐γ produced by various cells including T cells, NK cells, and dendritic cells was the primary cytokine for IgG2c CSR and B‐cell intrinsic MyD88 is required for IFN‐γ production. Moreover, IFN‐γ receptor (IFNγR) deficiency abolished sHZ‐induced IgG2c production, while recombinant IFN‐γ administration successfully rescued IgG2c CSR impairment in mice lacking B‐cell intrinsic MyD88. Together, our results show that B cell‐intrinsic MyD88 signaling is involved in the MOA of certain particulate adjuvants and this may enhance our specific understanding of how adjuvants and vaccines work. [ABSTRACT FROM AUTHOR]

Background and Aim: Immunoglobulin G4‐related sclerosing cholangitis (IgG4‐SC) presents as isolated proximal‐type sclerosing cholangitis (i‐SC). The present study sought to clarify the imaging differences between i‐SC and Klatskin tumor. Differences between i‐SC and IgG4‐SC associated with autoimmune pancreatitis (AIP‐SC) were also studied. Methods: Differentiating factors between i‐SC and Klatskin tumor were studied. Serum IgG4 level, CA19‐9 level, computed tomography (CT) findings, cholangiography findings (symmetrical smooth long stricture extending into the upper bile duct [SSLS]), endosonographic features (continuous symmetrical mucosal lesion to the hilar part [CSML]), endoscopic biopsy results, treatment, relapse, and survival were also compared between patients with i‐SC and those with AIP‐SC. Results: For a differential diagnosis between i‐SC (N = 9) and Klatskin tumor (N = 47), the cut‐off value of serum IgG4 level was 150 mg/dL (sensitivity, 0.857, specificity, 0.966). Logistic regression analysis indicated that serum IgG4 level, presence of SSLS, presence of CSML, and presence of swollen ampulla are independent factor for identifying i‐SC. Relapse rate was significantly higher in the IgG4‐SC with AIP group than in the i‐SC group (log rank, P = 0.046). Conclusion: Isolated proximal‐type sclerosing cholangitis presents as a nodular lesion with SSLS and/or CSML mimicking a Klatskin tumor. Those endoscopic features might provide a diagnostic clue for i‐SC. i‐SC is likely to have a more favorable prognosis than IgG4‐SC with AIP. [ABSTRACT FROM AUTHOR]

Neutrophil extracellular trap (NET) formation involves the release of DNA outside the cell to neutralize pathogens. Techniques such as live microscopy, flow cytometry, and intravital imaging allow the characterization of NETs, but these either cannot be applied in vivo, lack specificity or require invasive procedures. We developed an automated analysis method to rapidly acquire and characterize cells as NETs or NET precursors, as opposed to cells undergoing other forms of cell death, using imaging flow cytometry. NETs were maintained in solution using a novel three‐dimensional cell culture system in which cells are suspended at the interface of two liquids of different density. Critically, we identify NETs using an image analysis algorithm based on morphological data showing the extrusion of DNA beyond the cell boundaries. In vitro, we used this technique to demonstrate different requirements for NET formation in human and mouse neutrophils. We also measured NETs in whole blood during infection of mice with the malaria parasite Plasmodium yoelii. We expect this technique will provide a valuable approach to better understand the process of NET formation and its importance in disease. © 2019 International Society for Advancement of Cytometry [ABSTRACT FROM AUTHOR]

Summary: Elastomeric isolation bearings consist of multiple rubber layers with their top and bottom surfaces bonded to steel plates to restrict compressive deformation. Deformation constraints result in a variation of elastic modulus over the cross section of the rubber layers. In this paper, we describe a normalized compression modulus distribution on a circular rubber pad. The compressive and bending moduli of the rubber pad can be reproduced by applying the distribution to a series of axial springs. We also present a mechanical model for predicting the behavior of elastomeric seismic isolation bearings subject to large shear deformation and high compressive load. The mechanical model consists of a series of multiple shear springs at midheight and a series of axial springs at the top and bottom interfaces of the bearing. Simulation analyses of bearing tests were conducted to validate the proposed model. The analyses demonstrated that a model for circular lead‐rubber bearings can successfully capture the influence of the axial load magnitude on the bearing shear behavior. The new model can simulate much more realistic behavior than prior models based on a uniform modulus assumption. [ABSTRACT FROM AUTHOR]

An effective vaccine against Pseudomonas aeruginosa would be hugely beneficial to people who are susceptible to the serious infections it can cause. Vaccination against PcrV of the P. aeruginosa type III secretion system is a potential prophylactic strategy for improving the incidence and prognosis of P. aeruginosa pneumonia. Here, the effect of nasal PcrV adjuvanted with CpG oligodeoxynucleotide (CpG) was compared with a nasal PcrV/aluminum hydroxide gel (alum) vaccine. Seven groups of mice were vaccinated intranasally with one of the following: 1, PcrV‐CpG; 2, PcrV‐alum; 3, PcrV alone; 4, CpG alone; 5, alum alone; 6 and 7, saline control. Fifty days after the first immunization, anti‐PcrV IgG, IgA and IgG isotype titers were measured; significant increases in these titers were detected only in the PcrV‐CpG vaccinated mice. The vaccinated mice were then intratracheally infected with a lethal dose of P. aeruginosa and their body temperatures and survival monitored for 24 hr, edema, bacteria, myeloperoxidase activity and lung histology also being evaluated at 24 hr post‐infection. It was found that 73% of the PcrV‐CpG‐vaccinated mice survived, whereas fewer than 30% of the mice vaccinated with PcrV‐alum or adjuvant alone survived. Lung edema and other inflammation‐related variables were less severe in the PcrV‐CpG group. The significant increase in PcrV‐specific IgA titers detected following PcrV‐CpG vaccination is probably a component of the disease protection mechanism. Overall, our data show that intranasal PcrV‐CpG vaccination has potential efficacy for clinical application against P. aeruginosa pneumonia. [ABSTRACT FROM AUTHOR]

ABSTRACT The pathomechanism of the ligamentum flavum (LF) hypertrophy in diabetic patients with lumbar spinal canal stenosis (LSCS) remains unclear. A cross-sectional study was undertaken to investigate the mechanism of LF hypertrophy in these patients. Twenty-four diabetic and 20 normoglycemic patients with LSCS were enrolled in the study. The structure of the LF in the study subjects was evaluated using histological and immunohistochemical methods, and the levels of sorbitol, pro-inflammatory cytokines, and the fibrogenic factor, TGF-β1, in the LF were analyzed. In vitro experiments were performed using NIH3T3 fibroblasts to evaluate the effect of high-glucose conditions and an aldose reductase inhibitor on the cellular production of sorbitol, pro-inflammatory factors, and TGF-β1. We found that the LF of diabetic patients exhibited significantly higher levels of sorbitol and pro-inflammatory cytokines, TGF-β1 and of CD68-positive staining than that of the normoglycemic subjects. The diabetic LF was significantly thicker than that of the controls, and showed evidence of degeneration. The high glucose-cultured fibroblasts exhibited significantly higher levels of sorbitol, pro-inflammatory factors, and TGF-β1 compared to the low glucose-cultured cells, and these levels were dose-dependently reduced by treatment with the aldose reductase inhibitor. Taken together, our data suggests that increased sorbitol levels in the LF of diabetic patients results in increased production of pro-inflammatory and fibrogenic factor, which contribute to LF hypertrophy, and could increase the susceptibility of diabetic patients to LSCS. Furthermore, aldose reductase inhibition effectively reduced the levels of sorbitol and sorbitol-induced pro-inflammatory factor expression in high glucose-cultured fibroblasts. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:1058-1066, 2017. [ABSTRACT FROM AUTHOR]

This paper presents a mechanical model for predicting the behavior of elastomeric seismic isolation bearings subject to combined end rotations and shear deformation. The mechanical model consists of a series of axial springs at the top, mid-height and bottom of the bearing to vertically reproduce asymmetric bending moment distribution in the bearings. The model can take into account end rotations of the bearing, and the overall rotational stiffness includes the effect of the variation of vertical load on the bearing and the imposed shear deformation. Static bending tests under various combinations of vertical load and shear deformation were performed to identify the mechanical characteristics of bearings. The test results indicate that bearing rotational stiffness increases with increasing vertical load but decreases with increasing shear deformation. Simulation analyses were conducted to validate the new mechanical model. The results of analyses using the new model show very good agreement with experimental observations. A series of seismic response analyses were performed to demonstrate the dynamic behavior of top-of-column isolated structures, a configuration where the end rotations of isolation bearings are typically expected to be larger. The results suggest that the end rotations of elastomeric bearings used in practical top-of-column isolated structures do not reduce the stability limit of isolation system. Copyright © 2016 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

ABSTRACT Vaccination against the type III secretion system of P. aeruginosa is a potential prophylactic strategy for reducing the incidence and improving the poor prognosis of P. aeruginosa pneumonia. In this study, the efficacies of three different adjuvants, Freund's adjuvant (FA), aluminum hydroxide (alum) and CpG oligodeoxynucleotide (ODN), were examined from the viewpoint of inducing PcrV-specific immunity against virulent P. aeruginosa. Mice that had been immunized intraperitoneally with recombinant PcrV formulated with one of the above adjuvants were challenged intratracheally with a lethal dose of P. aeruginosa. The PcrV-FA immunized group attained a survival rate of 91%, whereas the survival rates of the PcrV-alum and PcrV-CpG groups were 73% and 64%, respectively. In terms of hypothermia recovery after bacterial instillation, PcrV-alum was the most protective, followed by PcrV-FA and PcrV-CpG. The lung edema index was lower in the PcrV-CpG vaccination group than in the other groups. PcrV-alum immunization was associated with the greatest decrease in myeloperoxidase in infected lungs, and also decreased the number of lung bacteria to a similar number as in the PcrV-FA group. There was less neutrophil recruitment in the lungs of mice vaccinated with PcrV-alum or PcrV-CpG than in those of mice vaccinated with PcrV-FA or PcrV alone. Overall, in terms of mouse survival the PcrV-CpG vaccine, which could be a relatively safe next-generation vaccine, showed a comparable effect to the PcrV-alum vaccine. [ABSTRACT FROM AUTHOR]

ABSTRACT Intervertebral disc degeneration proceeds with age and is one of the major causes of lumbar pain and degenerative lumbar spine diseases. However, studies in the field of intervertebral disc biology have been hampered by the lack of reliable cell lines that can be used for in vitro assays. In this study, we show that a chordoma-derived cell line U-CH1-N cells highly express the nucleus pulposus (NP) marker genes, including T (encodes T brachyury transcription factor), KRT19, and CD24. These observations were further confirmed by immunocytochemistry and flow cytometry. Reporter analyses showed that transcriptional activity of T was enhanced in U-CH1-N cells. Chondrogenic capacity of U-CH1-N cells was verified by evaluating the expression of extracellular matrix (ECM) genes and Alcian blue staining. Of note, we found that proliferation and synthesis of chondrogenic ECM proteins were largely dependent on T in U-CH1-N cells. In accordance, knockdown of the T transcripts suppressed the expression of PCNA, a gene essential for DNA replication, and SOX5 and SOX6, the master regulators of chondrogenesis. On the other hand, the CD24-silenced cells showed no reduction in the mRNA expression level of the chondrogenic ECM genes. These results suggest that U-CH1-N shares important biological properties with notochordal NP cells and that T plays crucial roles in maintaining the notochordal NP cell-like phenotype in this cell line. Taken together, our data indicate that U-CH1-N may serve as a useful tool in studying the biology of intervertebral disc. © 2016 The Authors. Journal of Orthopaedic Research Published by Wiley Periodicals, Inc. on behalf of Orthopaedic Research Society. J Orthop Res 34:1341-1350, 2016. [ABSTRACT FROM AUTHOR]

Paired immunoglobulin-like type 2 receptor α (PILRα) is an inhibitory receptor that is mainly expressed on myeloid cells, and negatively regulates neutrophil infiltration during inflammation. However, PILRα role on monocyte has not been described. Under both steady-state and inflammatory conditions, monocytes migrate into tissues and differentiate into macrophages. Macrophages in adipose and liver tissues play important roles in tissue homeostasis and pathogenesis of metabolic diseases. Here, we found that PILRα controls monocyte mobility through regulating integrin signaling and inhibiting CD99-CD99 binding. Moreover, we found that Pilra−/− mice developed obesity and hepatomegaly with fibrosis, and the numbers of macrophages in adipose and liver tissues are significantly increased in Pilra−/− mice. These data suggest that immune inhibitory receptor, PILRα, plays an important role in the prevention of obesity and liver fibrosis. [ABSTRACT FROM AUTHOR]

CpG oligodeoxynucleotide (ODN) is one of promising nucleic acid-based adjuvants. We recently improved its ability to enhance CD8+ T-cell responses to coadministered protein antigen without conjugation or emulsion, by forming a nanoparticulate complex between CpG ODN (K3) and mushroom-derived β-glucan schizophyllan (SPG), namely K3-SPG. Here, we sought to elucidate the cellular immunological mechanisms by which K3-SPG induce such potent CD8+ T-cell responses to coadministered antigen. By focusing on two DC subsets, plasmacytoid DCs and CD8α+ DCs, as well as the secreted cytokines, IFN-α and IL-12, we found that K3-SPG strongly activates mouse plasmacytoid DCs to secrete IFN-α and CD8α+ DCs to secrete IL-12, respectively. Although a single cytokine deficiency had no impact on adjuvant effects, the lack of both type I IFN and IL-12 in mice resulted in a significant reduction of Th1 type immune responses and CD8+ T-cell responses elicited by protein vaccine model. By sharp contrast, type I IFN, but not IL-12, was required for the production of IFN-γ by human PBMCs as well as antigen-specific CD8+ T-cell proliferation. Taken together, K3-SPG may overcome the species barrier for CpG ODN to enhance antigen-specific CD8+ T-cell responses despite the differential role of IL-12 between human and mice. [ABSTRACT FROM AUTHOR]

Cancer cells contain a small population of cancer stem cells or cancer initiating cells, which can be enriched in the side population ( SP) after fluorescence activated cell sorting. To examine the members of the ADAM, ADAMTS and MMP gene families related to phenotypes of the SP and the main population ( MP), we screened the expression of all the members in the propagated SP and MP of A549 lung adenocarcinoma cells, and found that the relative expression ratio of ADAM23 in the MP to the SP is most highly increased, but none of them are increased in the SP. A similar result on the ADAM23 expression was obtained with another cell line, Calu-3 cells. Overexpression of ADAM23 inhibited colony formation, cell adhesion and migration, and knockdown of ADAM23 by sh RNA showed the reverse effects. ADAM23-mediated suppression of colony formation, cell adhesion and migration was greatly reduced by treatment with neutralizing anti- ADAM23 antibody, anti-αvβ3 integrin antibody and/or ADAM23 disintegrin peptide. Expression of cancer stem cell-related genes, including AKRC1/2, TM4 SF1 and NR0B1, was increased by knockdown of ADAM23. In addition, lung metastasis of A549 transfectants with different levels of ADAM23 expression was negatively regulated by the ADAM23 expression levels. Our data provide evidence that ADAM23 plays a role in suppression of cancer cell progression through interaction with αvβ3 integrin, and suggest that downregulation of ADAM23 in SP cells may contribute toward providing a cancer stem cell phenotype by facilitating the activity of integrin αvβ3. [ABSTRACT FROM AUTHOR]

Prothymosin-alpha protects the brain and retina from ischemic damage. Although prothymosin-alpha contributes to toll-like receptor (TLR4)-mediated immnunopotentiation against viral infection, the beneficial effects of prothymosin-alpha-TLR4 signaling in protecting against ischemia remain to be elucidated. In this study, intravitreal administration of prothymosinalpha 48 h before induction of retinal ischemia prevented retinal cellular damage as evaluated by histology, and retinal functional deficits as evaluated by electroretinography. Prothymosin- alpha preconditioning completely prevented the ischemia- induced loss of ganglion cells with partial survival of bipolar and photoreceptor cells, but not amacrine cells, in immunohistochemistry experiments. Prothymosin-alpha treatment in the absence of ischemia caused mild activation, proliferation, and migration of retinal microglia, whereas the ischemia-induced microglial activation was inhibited by prothymosin-alpha preconditioning. All these preventive effects of prothymosin-alpha preconditioning were abolished in TLR4 knock-out mice and by pre-treatments with anti-TLR4 antibodies or minocycline, a microglial inhibitor. Prothymosinalpha preconditioning inhibited the retinal ischemia-induced up-regulation of TLR4-related injury genes, and increased expression of TLR4-related protective genes. Furthermore, the prothymosin-alpha preconditioning-induced prevention of retinal ischemic damage was abolished in TIR-domain-containing adapter-inducing interferon-p knock-out mice, but not in myeloid differentiation primary response gene 88 knock-out mice. Taken together, the results of this study suggest that prothymosin-alpha preconditioning selectively drives TLR4-TIR-domain-containing adapter-inducing interferon-p signaling and microglia in the prevention of retinal ischemic damage. [ABSTRACT FROM AUTHOR]

The development of cancer immunotherapies is progressing rapidly with a variety of technological approaches. They consist of “cancer vaccines”, which are based on the idea of vaccination, “effector cell therapy”, classified as passive immunotherapy, and “inhibition of immunosuppression”, which intends to break immunological tolerance to autoantigens or immunosuppressive environments characterizing antitumor immune responses. Recent reports showing clinical evidence of efficacy of immune checkpoint inhibitors and adoptive immunotherapies with tumor-infiltrating lymphocytes and tumor-specific receptor gene-modified T cells indicate the beginning of a new era for cancer immunotherapy. This guidance summarizes ideas that will be helpful to those who plan to develop cancer immunotherapy. The aims of this guidance are to discuss and offer important points in early phase clinical studies of innovative cancer immunotherapy, with future progress in this field, and to contribute to the effective development of cancer immunotherapy aligned with the scope of regulatory science. This guidance covers cancer vaccines, effector cell therapy, and inhibition of immunosuppression, including immune checkpoint inhibitors. [ABSTRACT FROM AUTHOR]

Numerous monoclonal antibodies ( mAb) targeting tumor antigens have recently been developed. Antibody-dependent cellular cytotoxicity ( ADCC) and antibody-dependent cellular phagocytosis ( ADCP) via effector cells such as tumor-infiltrating natural killer ( NK) cells and macrophages are often involved in mediating the antitumor activity of mAb. CpG oligodeoxynucleotides ( ODN) have a potent antitumor activity and are considered to increase tumor infiltration of NK cells and macrophages. Our group previously reported significant antitumor activity of anti-bone marrow stromal antigen 2 ( BST2) mAb against BST2-positive endometrial cancer cells through ADCC. In this study, we evaluated the synergistic antitumor activity of combination therapy with anti- BST-2 mAb and CpG ODN using SCID mice and elucidated the mechanisms underlying this activity. Anti- BST2 mAb and CpG ODN monotherapy had a significant dose-dependent antitumor activity ( P = 0.0135 and P = 0.0196, respectively). Combination therapy with anti- BST2 mAb and CpG ODN had a significant antitumor activity in SCID mice ( P < 0.01), but not in NOG mice. FACS analysis revealed significantly increased numbers of NK cells and macrophages in tumors treated with a combination of anti- BST2 mAb and CpG ODN and with CpG ODN alone in SCID mice ( P < 0.05 and P < 0.01, respectively). These results suggested that the combination therapy with anti- BST2 mAb and CpG ODN has a significant antitumor activity and induces tumor infiltration of NK cells and macrophages. Combination therapy with CpG ODN and anti- BST2 mAb or other antitumor mAb depending on ADCC may represent a new treatment option for cancer. [ABSTRACT FROM AUTHOR]

Agonists for TLR9 and Stimulator of IFN Gene (STING) act as vaccine adjuvants that induce type-1 immune responses. However, currently available CpG oligodeoxynucleotide (ODN) (K-type) induces IFNs only weakly and STING ligands rather induce type-2 immune responses, limiting their potential therapeutic applications. Here, we show a potent synergism between TLR9 and STING agonists. Together, they make an effective type-1 adjuvant and an anticancer agent. The synergistic effect between CpG ODN (K3) and STING-ligand cyclic GMP-AMP (cGAMP), culminating in NK cell IFN-γ (type-II IFN) production, is due to the concurrent effects of IL-12 and type-I IFNs, which are differentially regulated by IRF3/7, STING, and MyD88. The combination of CpG ODN with cGAMP is a potent type-1 adjuvant, capable of inducing strong Th1-type responses, as demonstrated by enhanced antigen-specific IgG2c and IFN-γ production, as well as cytotoxic CD8+ T-cell responses. In our murine tumor models, intratumoral injection of CpG ODN and cGAMP together reduced tumor size significantly compared with the singular treatments, acting as an antigen-free anticancer agent. Thus, the combination of CpG ODN and a STING ligand may offer therapeutic application as a potent type-II IFN inducer. [ABSTRACT FROM AUTHOR]

Autoimmune blistering diseases are organ-specific autoimmune diseases characterized by autoantibodies against structural proteins that maintain cell-cell (pemphigus diseases) and cell-matrix adhesions (pemphigoid diseases) in the skin and mucous membranes. Over the last few decades, identification of autoantigens and extensive characterization of autoantibodies have improved understanding of the pathogenesis of these diseases. In addition, the development of new sensitive and specific immunoassays enabled accurate diagnosis and proper evaluation of disease activity in autoimmune blistering diseases. In this review, we describe practical updates for molecular diagnostic tests for autoimmune blistering diseases and the basis for interpreting the results of the assays. [ABSTRACT FROM AUTHOR]

Reprogramming of toll-like receptor 4 ( TLR4) by brief ischemia or lipopolysacharide ( LPS) contributes to superintending tolerance against destructive ischemia in brain. However, beneficial roles of TLR4 signaling in ischemic retina are not well known. This study demonstrated that preconditioning with LPS 48 h prior to the retinal ischemia prevents the cellular damage in morphology with hematoxylin and eosin (H&E) staining and functions of retina with electroretinogram ( ERG), while post-ischemia treatment deteriorated it. The preventive effects of LPS preconditioning showed the cell type-specificity of retinal cells. There was complete rescue of ganglion cells, partial rescue of bipolar and photoreceptor cells or no rescue of amacrine cells, respectively. LPS treatment caused the proliferation and migration of retinal microglia and its preconditioning prevented the ischemia-induced microglial activation. Preventive actions from cell damages following LPS preconditioning prior to retinal ischemia were abolished in TLR4 knock-out mice, and by pre-treatments with anti- TLR4 antibody or minocycline, a microglia inhibitor, which themselves had no effects on the retinal ischemia-induced damages or microglia activation. Thus, this study revealed that TLR4 mediates the LPS preconditioning-induced preventive effects through microglial activation in the retinal ischemia model. [ABSTRACT FROM AUTHOR]

Several biomarkers have been used to evaluate osteoarthritis of the limb joints. Here we evaluated the use of serum cartilage metabolites as biomarkers for degenerative lumbar scoliosis (DLS). Fifty-two DLS patients with Cobb angle > 10° were compared with 19 control patients. Serum levels of hyaluronic acid (HA), keratan sulfate (KS), cartilage oligomeric matrix protein (COMP), collagen type II cleavage (C2C), and procollagen type II C-propeptide (CPII) were measured. Serum levels of KS (DLS 1.20 ± 0.44 µg/ml vs. control 0.98 ± 0.33 µg/ml), CPII (DLS 1905.1 ± 948.2 ng/ml vs. control 1223.6 ± 884.4 ng/ml), and C2C (DLS 219.1 ± 59.2 ng/ml vs. control 177.7 ± 71.7 ng/ml) were significantly higher in DLS. There were no significant differences in the levels of HA or COMP. There was a significant positive correlation between the Cobb angle and CPII in DLS. This is the first study to evaluate the cartilage biomarkers in DLS. The results suggest DLS patients have higher levels of type II collagen synthesis and degradation, indicated by elevated serum CPII and C2C, respectively. As type II collagen is a major component of collagens in the nucleus pulposus and facet joint cartilages, its enhanced turnover may be related to the development and progression of DLS. © 2012 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 30:1249-1253, 2012 [ABSTRACT FROM AUTHOR]

Autoantibodies in pemphigus foliaceus (PF) and vulgaris (PV) bind to desmoglein (Dsg) 1 and 3, respectively, and cause loss of keratinocyte adhesion. To characterize the pathogenicity and genetics of such antibodies we have used phage display to isolate monoclonal antibodies (mAbs) from patients. PCR is used to clone the heavy and light chain variable region of the peripheral B cells into a vector that creates a phage particle with the antibody expressed on its surface and the cDNA encoding that antibody inside. The library of phage produced from a PF or PV patient are then panned on a plate containing Dsg1 or Dsg3 to isolate clones. The cDNA of each clone is sequenced to characterize the genetics of the expressed mAb. The mAb from each unique clone is tested for pathogenicity either by injecting into normal human skin organ culture or into neonatal mice. Pathogenic antibodies cause typical pemphigus blisters. In both PV and PF patients the heavy chain (VH) genes used for Dsg-binding antibodies are severely restricted. PV and PF patients have both pathogenic and non-pathogenic mAbs. The immunochemical characteristics of the antibodies (including pathogenicity) sort with the VH, not the VL, gene. These monoclonal pathogenic antibodies can be used to screen peptide libraries to find short peptides that block antibody binding. In summary, the antibody response is restricted and, therefore, it may be feasible to target the specific pathogenic antibodies for therapy. [ABSTRACT FROM AUTHOR]

Introducing double-stranded DNA (dsDNA) into the cytoplasm of macrophages and dendritic cells triggers the activation of these professional antigen-presenting cells (APCs). This process is characterized by the up-regulation of costimulatory molecules and the production of various cytokines, chemokines, and antibacterial/viral factors. Current findings indicate that interferon-β (IFN-β) plays a key role in the stimulatory cascade triggered by dsDNA. Both immune and non-immune cells respond to intracytoplasmic dsDNA by up-regulating IFN-β) expression, a process that reduces host susceptibility to infection. The immune activation induced by dsDNA is independent of MyD88, TRIF and DNA-PKcs, indicating that a Toll-like receptor-independent mechanism underlies the cellular activation mediated by intracytoplasmic dsDNA. [ABSTRACT FROM AUTHOR]

We previously reported that the transplantation of neural stem/progenitor cells (NSPCs) can contribute to the repair of injured spinal cord in adult rats and monkeys. In some cases, however, most of the transplanted cells adhered to the cavity wall and failed to migrate and integrate into the host spinal cord. In this study we focused on chondroitin sulfate proteoglycan (CSPG), a known constituent of glial scars that is strongly expressed after spinal cord injury (SCI), as a putative inhibitor of NSPC migration in vivo. We hypothesized that the digestion of CSPG by chondroitinase ABC (C-ABC) might promote the migration of transplanted cells and neurite outgrowth after SCI. An in vitro study revealed that the migration of NSPC-derived cells was inhibited by CSPG and that this inhibitory effect was attenuated by C-ABC pre-treatment. Consistently, an in vivo study of C-ABC treatment combined with NSPC transplantation into injured spinal cord revealed that C-ABC pre-treatment promoted the migration of the transplanted cells, whereas CSPG-immunopositive scar tissue around the lesion cavity prevented their migration into the host spinal cord in the absence of C-ABC pre-treatment. Furthermore, this combined treatment significantly induced the outgrowth of a greater number of growth-associated protein-43-positive fibers at the lesion epicentre, compared with NSPC transplantation alone. These findings suggested that the application of C-ABC enhanced the benefits of NSPC transplantation for SCI by reducing the inhibitory effects of the glial scar, indicating that this combined treatment may be a promising strategy for the regeneration of injured spinal cord. [ABSTRACT FROM AUTHOR]

Conventional methods of gender recognition cannot be applied to counting store customers in terms of both data acquisition conditions and processing time. To solve this problem, the authors proposed a gender recognition method based on human silhouette shape characteristics. The current study demonstrated for the first time how efficient the footstep characteristic and foot pressure characteristic are for gender recognition. It discussed the recognition technique based on joining of the mentioned characteristics with the silhouette characteristic. These were joined by the one-dimensional compression based on the Fischer discrimination of characteristic vectors arranged in the order of characteristic rates for these three types. Based on the results of clothing-related research aimed at department store customers, a population group consisting of 45 male and 45 female participants was gathered to simulate distribution of various types of clothing and footwear. The authors conducted recognition tests with this population group and examined distributions by types of clothes and footwear in the characteristic space. The results showed that (1) for each separate characteristic rate the recognition rate which used to be 80% increased up to 90% due to use of joint characteristics, (2) there were numerous samples that featured correct recognition by the footstep characteristic or foot pressure characteristic even with erroneous recognition by the silhouette characteristic, which demonstrated effectiveness of the proposed combination of the three types of characteristic rates. © 2002 Wiley Periodicals, Inc. Electron Comm Jpn Pt 2, 85(8): 54–64, 2002; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/ecjb.10066 [ABSTRACT FROM AUTHOR]

1 The involvement of bradykinin (BK) B[sub 2] receptor in acute pancreatitis induced by pancreaticobiliary duct ligation was investigated in rats. 2 The activities of amylase and lipase in the serum, the water content of the pancreas, and vacuolization of the acinar cells were significantly increased 2 h after obstruction of the duct in Sprague-Dawley rats. 3 Elevated serum amylase activity, increased pancreatic oedema, and damage of the pancreatic tissue were significantly less marked in plasma kininogen-deficient, B/N-Katholiek rats than in the normal strain, B/N-Kitasato rats 2 h after the ligation. 4 Obstruction of the pancreaticobiliary duct augmented the level of (1-5)-BK (Arg¹-Pro²-Pro³-Gly[sup 4]Phe[sup 5]), a stable BK metabolite, in the blood from 73.0±21.7 pg ml[sup -1] at 0 h to 149.8 ± 38.0 pg ml[sup -1] at 2 h after the induction of pancreatitis in SD rats. 5 Administration of a BK B[sub 2] receptor antagonist, FR173657 (100 mg kg[sup -1], p.o.) or Hoel40 (100 nmol kg[sup -1,, s.c.), reduced the elevation of amylase and lipase activities in the serum and of pancreatic water content in a dose-dependent manner. The effective attenuation of oedema formation and vacuolization by the antagonists was also confirmed light-microscopically. In contrast, treatment with gabexate mesilate or indomethacin did not cause significant suppression of the pancreatitis. 6 These findings suggest a possible involvement of kinin B[sub 2] receptor in the present pancreatitis model. Furthermore, they point to the potential usefulness of the B[sub 2] receptor in clinical acute pancreatitis. [ABSTRACT FROM AUTHOR]

This paper proposes an automatic method of extracting the image of a vehicle moving along a straight road from a sequential video image. The method has been designed to be independent of the type of vehicle and the background, and to obtain an image with minimum defective areas. The image of the vehicle is extracted by analyzing a locus in a spatio-temporal image seen through a slit in a fixed video camera, by updating the background image and superimposing a time-series of the background difference image. The method can extract a full image of various types of vehicles in an arbitrary background with minimum defects, and can separate the images of two vehicles when one crosses in front of the other. The method has been tested successfully with actual traffic (including traffic starting from a traffic light). The results show that the method is practical fox smoothly Bowing traffic. The method has also been tested successfully for identifying vehicle types. [ABSTRACT FROM AUTHOR]

Assesses primary neural system/progenitor cells (NSPC) that were transplanted into injured spinal cord. Bioluminescence in the lentivirally transduced NSPC; Viability studies in spinal cord injury; Importance of the timing of NSPC transplantation to the function of integrated cells for the injured spinal cord.

Objective: Synthetic oligodeoxynucleotides (ODN) expressing TTAGGG motifs suppress the production of proinflammatory cytokines and have been proven effective at blocking the development of certain organ-specific autoimmune diseases. We undertook this study to determine whether suppressive ODN alter the development of systemic autoimmunity, by evaluating their effect on the progression of lupus-like disease in NZB x NZW (NZB/NZW) mice., Methods: We repeatedly treated mice with suppressive ODN before or after the onset of proteinuria. We monitored the effect of treatment on the onset, severity, and immunologic correlates of disease., Results: Treatment with suppressive ODN significantly prolonged lifespan while delaying the onset and progression of glomerulonephritis in NZB/NZW mice. Clinical improvement was accompanied by a significant reduction in anti-double-stranded DNA autoantibody production and by significantly reduced secretion of interferon-gamma and interleukin-12 in vivo., Conclusion: Suppressive ODN may be of benefit in the treatment of chronic systemic autoimmune diseases such as systemic lupus erythematosus.

Objective: To examine whether systemic administration of oligonucleotides (ODNs), known to inhibit the production of proinflammatory cytokines, alters host susceptibility to collagen-induced arthritis (CIA), a murine model of rheumatoid arthritis (RA)., Methods: CIA was induced by injecting DBA/1 mice with type II collagen (CII) in Freund's complete adjuvant, followed 3 weeks later by CII in Freund's incomplete adjuvant. The effect of suppressive ODNs on the incidence and severity of disease was monitored, as were immune correlates of CIA., Results: Suppressive ODNs administered during the inductive phase of CIA significantly reduced the incidence and severity of arthritis. Treatment with suppressive ODNs significantly decreased serum titers of pathogenic IgG anti-CII autoantibodies and interferon-gamma production by collagen-reactive T cells., Conclusion: Suppressive ODNs may be of therapeutic value in the treatment of RA, and potentially other autoimmune diseases.

Objective: To examine whether systemic administration of immunostimulatory and immunosuppressive oligodeoxynucleotides (ODNs) alter host susceptibility to inflammatory arthritis., Methods: Normal BALB/c mice were treated systemically with CpG ODNs or suppressive ODNs, and then challenged intraarticularly with CpG DNA. The onset and magnitude of the resulting inflammatory response was monitored., Results: Systemic delivery of CpG ODNs significantly increased susceptibility to local inflammation, whereas systemic treatment with suppressive ODNs reduced this susceptibility. CD11c+ cells played a key role in mediating host sensitivity to arthritis. These cells were the dominant source of tumor necrosis factor alpha production in CpG-stimulated animals and transferred resistance to arthritis from mice treated with suppressive ODNs., Conclusion: Systemic exposure to immunostimulatory and immunosuppressive DNA influences host susceptibility to local inflammatory challenge. Current findings raise the possibility that suppressive ODNs may be useful in the prevention/treatment of proinflammatory diseases.

Objective: Bacterial DNA contains immunostimulatory CpG motifs that cause inflammation when injected into the knee joints of normal mice. We examined whether synthetic oligodeoxynucleotides (ODN) that suppress CpG-induced immune responses prevent CpG-induced arthritis., Methods: CpG, suppressive, and/or control ODN were injected into the knees of BALB/c mice. Joint swelling and inflammation were evaluated by physical measurement, by histologic analysis of joint tissue, and by magnetic resonance imaging., Results: Immunostimulatory CpG DNA induced local arthritis, characterized by swelling of the knee joints, the presence of inflammatory cell infiltrates, the perivascular accumulation of mononuclear cells, and hyperplasia of the synovial lining. Administering suppressive (but not control) ODN reduced the manifestations and severity of arthritis up to 80%., Conclusion: Suppressive ODN may be useful for the prevention or treatment of arthritis induced by bacterial DNA.