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The octamer-binding transcriptionfactor 4 (Oct3/4) is a master gene in the transcriptional regulatorynetwork of pluripotent cells. Repression of Oct3/4 in embryonic stemcells (ESCs) is associated with cell differentiation and loss of pluripotency,whereas forced overexpression in cooperation with other transcriptionalfactors, such as Nanog, Sox2, and Lin28, can reprogram somatic cellsback into pluripotent cells, termed induced pluripotent stem cells(iPSCs). However, random integration and potential tumorigenic transformationcaused by viral transduction limit the clinical application of iPSCs.By performing a cell-based high throughput screening (HTS) campaign,we identified several potential small molecules as inducers of Oct3/4expression. Here we report a lead structure ethyl 2-((4-chlorophenyl)amino)-thiazole-4-carboxylate,termed O4I2, showing high activity in enforcing Oct3/4 expression.On the basis of chemical expansion, we further identified derivativeshaving increased activities toward Oct3/4 induction. Thus, O4I2 andits derivatives should provide a new class of small molecules suitablefor iPSC generation. [ABSTRACT FROM AUTHOR]

Two-dimensional (2D) coordination polymers consisting of MnIII Schiff-base complexes and dicarboxylic acids, [{Mn(salen)}4(L1)](PF6)2•(CH3OH)2 (C4; H2L1 = adipid acid) and [{Mn(salen)}4(L2)](PF6)2•(CH3OH)4 (C4′; H2L2 = E,E-1,3-butadiene-1,4-dicarboxylic acid) (salen2- = N,N′-(ethylene)bis(salicylideneiminato), were synthesized by using a one-pot reaction and characterized by using single-crystal X-ray crystallographic analysis. One-dimensional (1D) chains composed of Mn(salen) dimers, [Mn2], bridged by carboxylato ligands (−[Mn2]-OCO--[Mn2]−), were linked by dicarboxylato ligands with n-butyl (−C4H8−) (C4) and butadienyl aliphatic groups (−C4H4−) (C4′). From static magnetic measurements on both C4 and C4′, there were ferromagnetic interactions between the MnIII ions through the phenoxo oxygen atoms of the salen2-, and antiferromagnetic interactions between the MnIII ions through carboxylato ligands (−OCO−). As a result, weak ferromagnetism occurred because of the zigzag-shaped chain structure of C4 and C4′, and magnetic anisotropy for Mn(salen). In the magnetization curves for C4′, weak interchain interactions (Jlinker) occurred through the π-conjugated butadienyl linkers in C4′, which C4 did not have. In other words, changing from saturated to unsaturated aliphatic groups in the dicarboxylic acid linkers resulted in weak interactions between 1D-magnetic chain moieties. Therefore, in the case of only C4′, antiferromagnetic phase transition appeared at 2.3 K. Both coordination polymers exhibited slow relaxation of the magnetizations, which originated from SCM moieties, because C4 and C4′ showed magnetic correlations. It is noteworthy that alternating current (ac) susceptibilities for C4′ are frequency-dependent around the Néel temperature. From analysis of the ac susceptibilities for C4, α (dispersion coefficient of the relaxation of magnetization) varied linearly with 1/T. This signifies that C4 behaved as an SCM with a single relaxation process. On the other hand, in α versus 1/T plots for C4′, an inflection point was observed at the Néel temperature, indicating that Jlinkers had an effect on the distribution of the relaxation times. Moreover, the inflection point for C4′ disappeared when a dc magnetic field was applied. This is the first report showing a direct correlation between an antiferromagnetic phase transition and slow magnetic relaxation. [ABSTRACT FROM AUTHOR]

Two-dimensional (2D) coordination polymers consisting of MnIII Schiff-base complexes and dicarboxylic acids, [{Mn(salen)}4(L1)](PF6)2·(CH3OH)2 (C4; H2L1 = adipid acid) and [{Mn(salen)}4(L2)](PF6)2·(CH3OH)4 (C4′; H2L2 = E,E-1,3-butadiene-1,4-dicarboxylic acid) (salen2- = N,N′-(ethylene)bis(salicylideneiminato), were synthesized by using a one-pot reaction and characterized by using single-crystal X-ray crystallographic analysis. One-dimensional (1D) chains composed of Mn(salen) dimers, [Mn2], bridged by carboxylato ligands (−[Mn2]-OCO--[Mn2]−), were linked by dicarboxylato ligands with n-butyl (−C4H8−) (C4) and butadienyl aliphatic groups (−C4H4−) (C4′). From static magnetic measurements on both C4 and C4′, there were ferromagnetic interactions between the MnIII ions through the phenoxo oxygen atoms of the salen2-, and antiferromagnetic interactions between the MnIII ions through carboxylato ligands (−OCO−). As a result, weak ferromagnetism occurred because of the zigzag-shaped chain structure of C4 and C4′, and magnetic anisotropy for Mn(salen). In the magnetization curves for C4′, weak interchain interactions (Jlinker) occurred through the π-conjugated butadienyl linkers in C4′, which C4 did not have. In other words, changing from saturated to unsaturated aliphatic groups in the dicarboxylic acid linkers resulted in weak interactions between 1D-magnetic chain moieties. Therefore, in the case of only C4′, antiferromagnetic phase transition appeared at 2.3 K. Both coordination polymers exhibited slow relaxation of the magnetizations, which originated from SCM moieties, because C4 and C4′ showed magnetic correlations. It is noteworthy that alternating current (ac) susceptibilities for C4′ are frequency-dependent around the Néel temperature. From analysis of the ac susceptibilities for C4, α (dispersion coefficient of the relaxation of magnetization) varied linearly with 1/T. This signifies that C4 behaved as an SCM with a single relaxation process. On the other hand, in α versus 1/T plots for C4′, an inflection point was observed at the Néel temperature, indicating that Jlinkers had an effect on the distribution of the relaxation times. Moreover, the inflection point for C4′ disappeared when a dc magnetic field was applied. This is the first report showing a direct correlation between an antiferromagnetic phase transition and slow magnetic relaxation. [ABSTRACT FROM AUTHOR]

Background Symptomatic knee joint effusion is frequently observed after hip fracture, which may lead to postoperative knee pain during rehabilitation after hip fracture surgery. However, unfortunately, very little has been reported on this phenomenon in the literature. The purpose of the current study was to investigate the relationship between symptomatic knee effusion and postoperative knee pain and to clarify the reason of the effusion accompanied by hip fracture. Methods A total of 100 patients over 65 years of age with an acute hip fracture after fall were prospectively followed up. Knee effusion was assessed on admission and at the operating room before the surgery. If knee effusion was observed at the time of the surgery, synovial fluid was collected into syringes to investigate the cause of the effusion using a compensated polarized light microscope. Furthermore, for each patient, we evaluated age, sex, radiographic knee osteoarthritis (OA), type of the fracture, laterality, severity of the fracture, and postoperative knee pain during rehabilitation. These factors were compared between patients with and without knee effusion at the time of the surgery. As a statistical analysis, we used Mann--Whitney U-test for patients' age and categorical variables were analyzed by chi- square test or Fisher's exact test. Results A total of 30 patients presented symptomatic knee effusion at the time of the surgery. In patients with knee effusion, numbers of intertrochanteric fracture, radiographic knee OA, and postoperative knee pain were significantly large compared to those without effusion. In terms of synovial fluid analysis, calcium pyrophosphate dihydrate crystals were observed in 80% of patients with knee effusion. Conclusion From our study, approximately 63% of patients with knee effusion at the time of the surgery had postoperative knee pain. In addition, this effusion was basically related to pseudogout. [ABSTRACT FROM AUTHOR]

Methamidophos, a representative organophosphate insecticide, is regulated because of its severe neurotoxicity, but it is suspected of contaminating agricultural foods in many countries due to illicit use. To reveal unknown effects of methamidophos on human health, we evaluated the developmental immunotoxicity of methamidophos using a respiratory syncytial virus (RSV) infection mouse model. Pregnant mice were exposed to methamidophos (10 or 20 ppm) in their drinking water from gestation day 10 to weaning on postnatal day 21. Offsprings born to these dams were intranasally infected with RSV.The levels of interleukin-6 (IL-6) and interferon-gamma in the bronchoalveolar lavage fluids after infection were significantly decreased in offspring mice exposed to methamidophos. Treatment with methamidophos did not affect the pulmonary viral titers but suppressed moderately the inflammation of lung tissues of RSV-infected offspring, histopathologically.DNA microarray analysis revealed that gene expression of the cytokines in the lungs of offspring mice exposed to 20 ppm of methamidophos was apparently suppressed compared with the control. Methamidophos did not suppress IL-6 production in RSV-infected J774.1 cell cultures. Thus, exposure of the mother to methamidophos during pregnancy and nursing was suggested to cause an irregular immune response in the lung tissues in the offspring mice. [ABSTRACT FROM AUTHOR]

In order to develop a more effective self-etching primer, it is helpful to understand how a functional monomer conditions the surface of the teeth.In this study, the reactivity by the carboxylic acid in the N-methacryloyl glycine (NMGly) on the calcium phosphate in hydroxyapatite or dentin was studied. The efficacy of the NMGly as a functional monomer for a self-etching primer was then examined.Applying NMGly to both the enamel and dentin resulted in an increase in the bond strength of resin, since the carboxylic acid in the NMGly decalcified the calcium phosphate in the hydroxyapatite and dentin. The bond strength to dentin achieved was higher than with the enamel. This difference was most likely due to different enamel and dentin adhesion mechanisms. [Copyright &y& Elsevier]

Conventional dendritic cells (cDCs) are required for peripheral T cell homeostasis in lymphoid organs, but the molecular mechanism underlying this requirement has remained unclear. We here show that T cell--specific CD47- deficient (Cd47ΔT) mice have a markedly reduced number of T cells in peripheral tissues. Direct interaction of CD47-deficient T cells with cDCs resulted in activation of the latter cells, which in turn induced necroptosis of the former cells. The deficiency and cell death of T cells in Cd47ΔT mice required expression of its receptor signal regulatory protein α on cDCs. The development of CD4+ T helper cell--dependent contact hypersensitivity and inhibition of tumor growth by cytotoxic CD8+ T cells were both markedly impaired in Cd47ΔT mice. CD47 on T cells thus likely prevents their necroptotic cell death initiated by cDCs and thereby promotes T cell survival and function. [ABSTRACT FROM AUTHOR]

Multi-walled carbon nanotubes (MWCNTs), a kind of nanomaterial, are widely used in battery electrodes and composite materials, but the adverse effects associated with their accumulation in the living body have not been sufficiently investigated. MWCNTs are a fibrous material with molecules similar to asbestos fibers, and there are concerns about its effects on the respiratory system. In this study, we conducted a risk assessment by exposing mice using a previously developed nanomaterial inhalation exposure method. We quantified the exposure in the lungs by a lung burden test, evaluated the deterioration due to pneumonia using respiratory syncytial virus (RSV) infection, and measured inflammatory cytokines in bronchoalveolar lavage fluid (BALF). As a result, in the lung burden test, the amount of MWCNT in the lung increased according to the inhalation dose. In the RSV infection experiment, CCL3, CCL5, and TGF-ß, which are indicators of inflammation and lung fibrosis, were elevated in the MWCNTexposed group. Histological examination revealed cells phagocytosing MWCNT fibers. These phagocytic cells were also seen during the recovery period from RSV infection. The present study found that MWCNT remained in the lungs for about a month or more, suggesting that the fibers may continue to exert immunological effects on the respiratory system. Furthermore, the inhalation exposure method enabled the exposure of nanomaterials to the entire lung lobe, allowing a more detailed evaluation of the effects on the respiratory system. [ABSTRACT FROM AUTHOR]

EBI3 functions as the subunit of immune-regulatory cytokines, such as IL-27 and IL-35, by pairing with p28 and p35, respectively. We treated wild-type and EBI3-deficient mice with intratracheal administration of LPS and obtained bronchoalveolar lavage fluid (BALF) 24 h later. Although neutrophils were the predominant cells in BALF from both groups of mice, eosinophils were highly enriched and there was increased production of eosinophil-attracting chemokines CCL11 and CCL24 in BALF of EBI3-deficient mice. The bronchial epithelial cells and alveolar macrophages were the major producers of CCL11 and CCL24. Because no such increases in eosinophils were seen in BALF of p28/IL-27-deficient mice or WSX-1/IL-27Ra subunit-deficient mice upon intratracheal stimulation with LPS, we considered that the lack of IL-35 was responsible for the enhanced airway eosinophilia in EBI3-deficient mice. In vitro, IL-35 potently suppressed production of CCL11 and CCL24 by human lung epithelial cell lines treated with TNF-a and IL-1b. IL-35 also suppressed phosphorylation of STAT1 and STAT3 and induced suppressor of cytokine signaling 3. In vivo, rIL-35 dramatically reduced LPS-induced airway eosinophilia in EBI3-deficient mice, with concomitant reduction of CCL11 and CCL24, whereas neutralization of IL-35 significantly increased airway eosinophils in LPS-treated wild-type mice. Collectively, our results suggest that IL-35 negatively regulates airway eosinophilia, at least in part by reducing the production of CCL11 and CCL24. [ABSTRACT FROM AUTHOR]

Sialic acid-binding immunoglobulin-like lectins (Siglecs) are a family of cell-surface immune receptors that bind to sialic acid at terminal glycan residues. Siglecs also recognize nonsialic acid ligands, many of which remain to be characterized. Here, we found that Siglec5 and Siglec14 recognize lipid compounds produced by Trichophyton, a fungal genus containing several pathogenic species. Biochemical approaches revealed that the Siglec ligands are fungal alkanes and triacylglycerols, an unexpected finding that prompted us to search for endogenous lipid ligands of Siglecs. Siglec5 weakly recognized several endogenous lipids, but the mitochondrial lipid cardiolipin and the anti-inflammatory lipid 5-palmitic acid-hydroxystearic acid exhibited potent ligand activity on Siglec5. Further, the hydrophobic stretch in the Siglec5 N terminus region was found to be required for efficient recognition of these lipids. Notably, this hydrophobic stretch was dispensable for recognition of sialic acid. Siglec5 inhibited cell activation upon ligand binding, and accordingly, the lipophilic ligands suppressed interleukin-8 (IL-8) production in Siglec5-expressing human monocytic cells. Siglec14 and Siglec5 have high sequence identity in the extracellular region, and Siglec14 also recognized the endogenous lipids. However, unlike Siglec5, Siglec14 transduces activating signals upon ligand recognition. Indeed, the endogenous lipids induced IL-8 production in Siglec14-expressing human monocytic cells. These results indicated that Siglec5 and Siglec14 can recognize lipophilic ligands that thereby modulate innate immune responses. To our knowledge, this is the first study reporting the binding of Siglecs to lipid ligands, expanding our understanding of the biological function and importance of Siglecs in the innate immunity. [ABSTRACT FROM AUTHOR]

Objectives: Lactoferrin (LF) and lactoperoxidase (LPO) are present in human saliva. LF has been demonstrated to show antibacterial and antiviral activities. In saliva, LPO catalyzes the hydrogen peroxide-dependent oxidation of thiocyanate to hypothiocyanite that exhibits antimicrobial and antiviral properties. A randomized, open-label, parallelgroup clinical trial was conducted to examine the effectiveness of sucking tablets containing LF and LPO (LF+LPO) in alleviating symptoms of the common cold and/ or influenza infection. Methods: A total of 407 subjects were randomized into two groups, treatment and non-treatment groups, and each group was further classified into subgroups habitually wearing a face mask, washing their hands, or gargling. The common cold, influenza, and gastrointestinal symptoms were used to evaluate the effectiveness, and the incidence and duration of symptoms were statistically analyzed. Results: The incidence and duration of common cold, gastrointestinal symptoms, and influenza infection were not statistically different between treatment and non-treatment groups. LF+LPO tablets were moderately effective in reducing the incidence and duration of common cold symptoms in the subgroup that did not gargle and especially to shorten significantly the duration of fever higher than 38°C in the subgroup that did not wear a face mask. Conclusion: The results suggested that the effect of ingestion of the tablet is not obvious in alleviating common cold symptoms but may be helpful when the subjects do not follow precautionary measures such as gargling and the use of a protective face mask.en] [ABSTRACT FROM AUTHOR]

IL-27 is an immunoregulatory cytokine consisting of p28 and EBI3. Its receptor also has two subunits, WSX1 and gp130. Although IL-27 promotes Th1 differentiation in naive T cells, it also induces IL-10 expression in effector Th1 cells to curtail excessive immune responses. By using p28-deficient mice and WSX1-deficient mice (collectively called IL-27-deficient mice), we examined the role of IL-27 in primary infection by murine g-herpesvirus 68 (MHV68), a murine model of EBV. Upon airway infection with MHV68, IL-27-deficient mice had more aggravated lung inflammation than wild-type mice, although MHV68 infection per se was better controlled in IL-27-deficient mice. Although epithelial cells and alveolar macrophages were primarily infected by MHV68, interstitial macrophages and dendritic cells were the major producers of IL-27. The lung inflammation of IL-27-deficient mice was characterized by more IFN-γ-producing CD8+ T cells and fewer IL-10-producing CD8+ T cells than that of wild-type mice. An infectious mononucleosis-like disease was also aggravated in IL-27-deficient mice, with prominent splenomegaly and severe hepatitis. Infiltration of IFN-γ-producing effector cells and upregulation of the CXCR3 ligand chemokines CXCL9, CXCL10, and CXCL11 were noted in the liver of MHV68-infected mice. Oral neomycin effectively ameliorated hepatitis, with decreased production of these chemokines in the liver, suggesting that the intestinal microbiota plays a role in liver inflammation through upregulation of these chemokines. Collectively, IL-27 is essential for the generation of IL-10-producing effector cells in primary infection by MHV68. Our findings may also provide new insight into the mechanism of hepatitis associated with infectious mononucleosis. [ABSTRACT FROM AUTHOR]

To investigate the effects of perinatal exposure to tetrabromobisphenol A (TBBPA), a brominated flame retardant, on the immune system, a respiratory syncytial virus (RSV) infection mouse model was utilized. Female mice were exposed to TBBPA mixed with the diet from 10 days after conception to weaning on postnatal day 21. Offspring mice were infected intranasally with A2 strain of RSV. Although no general toxicological sign was observed, the pulmonary viral titers of offspring mice exposed to 0.1% TBBPA were significantly increased compared with the control on day 5 post-infection. TBBPA did not affect RSV growth in vitro. Histopathological analysis confirmed that the exacerbation of interstitial pneumonia was due to TBBPA- exposure in the lung tissues in RSV-infected offspring. Moreover, gene expression of interleukin (IL)-24 was shown to be elevated typically in the lung tissues of TBBPA- treated offspring by a DNA microarray and was also confirmed by immunohistopathological analysis using an anti-IL-24 antibody. Thus, developmental exposure to TBBPA affected the immune response to RSV infection, resulting in the exacerbation of pneumonia. Thus, IL-24 should be a key molecule to understand the mechanism of action of TBBPA. [ABSTRACT FROM AUTHOR]

Background: In patients with severe hemophilia A, standard treatment is regular prophylactic and episodic intravenous infusions of factor VIII. However, these treatments are burdensome, especially for children, and may lead to the formation of anti-factor VIII alloantibodies (factor VIII inhibitors). Emicizumab (ACE910), a humanized bispecific antibody mimicking the cofactor function of factor VIII, was developed to abate these problems.Methods: We enrolled 18 Japanese patients with severe hemophilia A (with or without factor VIII inhibitors) in an open-label, nonrandomized, interindividual dose-escalation study of emicizumab. The patients received subcutaneous emicizumab weekly for 12 weeks at a dose of 0.3, 1.0, or 3.0 mg per kilogram of body weight (cohorts 1, 2, and 3, respectively). The end points were safety and pharmacokinetic and pharmacodynamic profiles. An additional, exploratory end point was the annualized bleeding rate, calculated as 365.25 times the number of bleeding episodes, divided by the number of days in the treatment period as compared with the 6 months before enrollment.Results: Emicizumab was associated with neither serious adverse events nor clinically relevant coagulation abnormalities. Plasma concentrations of emicizumab increased in a dose-dependent manner. Activated partial-thromboplastin times remained short throughout the study. The median annualized bleeding rates in cohorts 1, 2, and 3 decreased from 32.5 to 4.4, 18.3 to 0.0, and 15.2 to 0.0, respectively. There was no bleeding in 8 of 11 patients with factor VIII inhibitors (73%) and in 5 of 7 patients without factor VIII inhibitors (71%). Episodic use of clotting factors to control bleeding was reduced. Antibodies to emicizumab did not develop.Conclusions: Once-weekly subcutaneous administration of emicizumab markedly decreased the bleeding rate in patients who had hemophilia A with or without factor VIII inhibitors. (Funded by Chugai Pharmaceutical; JapicCTI number, 121934.). [ABSTRACT FROM AUTHOR]

Murine experimental autoimmune uveitis (EAU) is a model for human autoimmune uveitis, whose pathogenesis is caused by both Th1 and Th17 cell responses. Epstein-Barr virus-induced gene 3 (EBI3) is a component of the heterodimeric cytokines: interleukin (IL)-27 and IL-35. Although IL-27 was shown to initiate Th1 cell development, it is also recognized as a negative regulator of fully activated CD4+ T cells, including Th17 cells. Recently, IL-35 also has also been reported to play immunosuppressive roles in autoimmunity. To investigate the roles of EBI3 in EAU, EBI3-/- mice were immunized with human interphotoreceptor retinoid binding protein peptide 1-20 (IRBP) to induce EAU. We observed that the clinical score in EBI3-/- mice was diminished compared with that in EBI3+/+ mice up to day 22 after immunization, whereas the score in EBI3-/- mice reached the same levels as that of EBI3+/+ mice after day 28. Histological analysis revealed a significant reduction of cellular infiltration into the retina in EBI3-/- mice on day 16. Although Th1 cell responses and IRBP-specific IL-10 production were reduced in EBI3-/- mice, the development of Th17 cell responses was unaffected on day 9. On day 21, Th1 cell responses and IRBP-specific IL-10 production was restored to the same levels as that in EBI3+/+ mice, and Th17 cell responses significantly increased in EBI3-/- mice. Furthermore, Foxp3 expression in CD4+ T cells was comparable between EBI3+/+ and EBI3-/- mice on days 9 and 21. Therefore, these results indicate that EBI3 may be important in EAU initiation by Th1 cell responses and may suppress EAU by inhibition of both Th1 and Th17 cell responses in the late/maintenance phase. [ABSTRACT FROM AUTHOR]

Chronic inflammation in arterial wall that is driven by immune cells and cytokines plays pivotal roles in the development of atherosclerosis. Interleukin 27 (IL-27) is a member of the IL-12 family of cytokines that consists of IL-27p28 and Epstein-Barr virus induced gene 3 (EBI3) and has anti-inflammatory properties that regulate T cell polarization and cytokine production. IL-27-deficient (Ldlr﹣/﹣Ebi3﹣/﹣) and IL-27 receptor-deficient (Ldlr﹣/﹣WSX-1﹣/﹣) Ldlr﹣/﹣ mice were generated and fed with a high-cholesterol diet to induce atherosclerosis. Roles of bone marrow-derived cells in vivo and macrophages in vitro were studied using bone marrow reconstitution by transplantation and cultured peritoneal macrophages, respectively. We demonstrate that mice lacking IL-27 or IL-27 receptor are more susceptible to atherosclerosis compared with wild type due to enhanced accumulation and activation of macrophages in arterial walls. The number of circulating proinflammatory Ly6Chi monocytes showed no significant difference between wild-type mice and mice lacking IL-27 or IL-27 receptor. Administration of IL-27 suppressed the development of atherosclerosis in vivo and macrophage activation in vitro that was indicated by increased uptake of modified low-density lipoprotein and augmented production of proinflammatory cytokines. These findings define a novel inhibitory role for IL-27 in atherosclerosis that regulates macrophage activation in mice. [ABSTRACT FROM AUTHOR]

In this article, the authors focus on the impact of oral administration of probiotics from Mongolian dairy products on the immune response in T helper type-1 (Th1) cytokine production in mouse. They inform that Lactobacillus plantarian 06CC2 strain increased the level of interleukin (IL)-12p40 in mouse cells co-culture with J774.1 cells. They mention that oral administration of the strain elevated the gene expression of interferon-gamma and IL-12p40.

Tetrabromobisphenol A (TBBPA), a brominated flame retardant, has been found to exacerbate pneumonia in respiratory syncytial virus- (RSV-) infected mice. We examined the effect of Brazilian propolis (AF-08) on the exacerbation of RSV infection by TBBPA exposure in mice. Mice were fed a powdered diet mixed with 1% TBBPA alone, 0.02% AF-08 alone, or 1% TBBPA and 0.02% AF-08 for four weeks and then intranasally infected with RSV. TBBPA exposure increased the pulmonary virus titer and level of IFN-γ, a representative marker of pneumonia due to RSV infection, in the lungs of infected mice without toxicity. AF-08 was significantly effective in reducing the virus titers and IFN-γ level increased by TBBPA exposure. Also, AF-08 significantly reduced proinflammatory cytokine (TNF-α and IL-6) levels in the lungs of RSV-infected mice with TBBPA exposure, but Th2 cytokine (IL-4 and IL-10) levels were not evidently increased. Neither TBBPA exposure nor AF-08 treatment affected the anti-RSV antibody production in RSV-infected mice. In flow cytometry analysis, AF-08 seemed to be effective in reducing the ratio of pulmonary CD8a+ cells in RSV-infected mice with TBBPA exposure. TBBPA and AF-08 did not exhibit anti-RSV activity in vitro. Thus, AF-08 probably ameliorated pneumonia exacerbated by TBBPA exposure in RSV-infected mice by limiting excess cellular immune responses. [ABSTRACT FROM AUTHOR]

WSX-1 is a subunit of the IL-27R, which plays a critical role in the initiation of Th1 responses. Murine experimental autoimmune uveitis (EAU) is a model of human autoimmune uveitis, in which a Th1 response predominates in the pathogenetic process. To explore the role of WSX-1 in this model, WSX-1−/− mice were immunized with interphotoreceptor retinoid-binding protein peptide 1–20 to induce EAU. We found that the EAU clinical and histological scores were lower in the WSX-1−/− mice up to day 21, whereas after day 21, the EAU scores were the same between the wild-type (WT) and WSX-1−/− mice with both declining at the same rate. In contrast to T lymphocytes from WT mice, WSX-1−/− T lymphocytes on day 9 after immunization failed to produce IFN-γ. Similarly, expression of Th1-related chemokines, such as regulated on activation, normal T cell expressed and secreted and IP-10, in the eye was reduced in WSX-1−/− mice compared with WT mice on day 13 after immunization. In addition, sub-retinal transfer of lymphocytes from WSX-1−/− mice on day 9 after immunization did not induce EAU in the recipient mice. Importantly, IFN-γ production, chemokine expression and the transferability of disease by lymphocytes became comparable for WSX-1−/− and WT mice at later stages. Thus, IL-27/WSX-1 affects the early development of EAU, and might be a target for therapy during the onset of autoimmune uveitis in humans. [ABSTRACT FROM AUTHOR]

To investigate the effects of the recombinant immunotoxin dsFv anti‐FRβ‐PE38, which consists of the disulfide‐stabilized Fv fragment (dsFv) of the anti–folate receptor β (anti‐FRβ) antibody and the 38‐kd portion of Pseudomonas exotoxin A (PE38), on the activation and proliferation of cells that function in inflammatory and degradative processes in rheumatoid arthritis (RA) synovial tissue.The Ig VH‐PE38 fusion protein and the Ig VL protein were produced in Escherichia coli, and then joined with a disulfide bond by engineering cysteine residues in the framework regions of these proteins. The effects of dsFv anti‐FRβ‐PE38 on the activation and proliferation of cells in RA synovial tissue were investigated by immunohistochemistry; the numbers of cells expressing CD68, vascular cell adhesion molecule 1, angiopoietin 1, CD34, proliferating cell nuclear antigen, and interleukin‐6 and the numbers of apoptotic cells were counted in RA synovial tissue engrafted into SCID mice treated or not treated with dsFv anti‐FRβ‐PE38. The effects of dsFv anti‐FRβ‐PE38 on the generation of osteoclasts from RA adherent synovial mononuclear cells in vitro was investigated by counting the number of resorption pits on dentin slices treated or not treated with dsFv anti‐FRβ‐PE38.Administration of dsFv anti‐FRβ‐PE38 reduced the numbers of macrophages, activated fibroblast‐like cells, endothelial cells, and proliferating cells and increased the numbers of apoptotic cells in RA synovial tissue engrafted into SCID mice. In vitro, the generation of osteoclasts from RA adherent synovial mononuclear cells was largely suppressed by treatment with dsFv anti‐FRβ‐PE38.Our findings show that dsFv anti‐FRβ‐PE38 immunotoxin would be a promising tool for the treatment of RA synovitis, especially when administered intraarticularly. [ABSTRACT FROM AUTHOR]

Objectives. Human endogenous uveitis is one of the sight-threatening diseases associated with variety of systemic disorders, such as Behcets disease and sarcoidosis. Recently, biosynthesized antibodies against inflammatory cytokines have been recognized to be useful to control the regional inflammation. In this study, we focused on the possibility of IL-6-based biological therapies for endogenous uveitis. We initially confirmed the significant increase of several inflammatory soluble factors including IL-6 in the vitreous fluids from refractory/chronic engogenous uveitis patients. Methods. To investigate the role of IL-6 in the formation of refractory ocular inflammation, we used the mouse experimental autoimmune uveitis (EAU) model. Both IL-6 and IL-23 are required for the development of IL-17-producing helper T subset (Th17) from naïve CD4+ T cells. Results. In the EAU model, neither IL-6-deficient mice nor IL-23-deficient mice could induce Th17 cells and the EAU score was decreased in these mice in the entire time course. We also confirmed that systemic administration of anti-IL-6 receptor antibody ameliorates EAU by suppressing both systemic and regional Th17 responses. Conclusions. IL-6 is responsible for causing ocular inflammation, and it is, at least partially, due to IL-6-dependent Th17 differentiation. IL-6 may be a target for therapy of refractory endogenous uveitis in humans. [ABSTRACT FROM AUTHOR]

IL-17-producing CD4+ T cells, so called Th17 cells, constitute a newly identified inflammatogenic cell population, which is critically involved in some inflammatory diseases. To explore the role of Th17 cells in murine experimental autoimmune uveoretinitis (EAU), a model of human autoimmune uveitis where Th1 responses predominantly participate in the pathogenesis, IL-17−/− mice were immunized with interphotoreceptor retinoid-binding protein peptide 1–20 for disease induction. Funduscopic examination revealed that EAU was induced in IL-17−/− mice just like in wild-type (WT) mice at early phases of the disease. However, at later/maintenance phases, the severity was significantly reduced in IL-17−/− mice. Expression of IFN-γ and MCP-1 was comparable between WT and IL-17−/− mice during the time course. In vivo blockade of IFN-γ and IL-4 resulted in exacerbation of EAU at later phases with augmented IL-17 production. Taken together, our data demonstrated that IL-17/Th17 participates in the late phases of EAU and also that Th1 and Th17 responses are differentially required for EAU. [ABSTRACT FROM AUTHOR]