グリオーマ幹細胞に対する免疫チェックポイント阻害薬とワクチン療法の併用

科学研究費助成事業 研究成果報告書：基盤研究(C)2015-2017課題番号 : 15K1032

Molecular and virulence characteristics of an outer membrane-associated RTX exoprotein in Pasteurella pneumotropica

<p>Abstract</p> <p>Background</p> <p><it>Pasteurella pneumotropica </it>is a ubiquitous bacterium that is frequently isolated from laboratory rodents and causes various clinical symptoms in immunodeficient animals. Currently two RTX toxins, PnxIA and PnxIIA, which are similar to hemolysin-like high-molecular-weight exoproteins are known in this species. In this study, we identified and analyzed a further RTX toxin named PnxIIIA and the corresponding type I secretion system.</p> <p>Results</p> <p>The RTX exoprotein, PnxIIIA, contains only a few copies of the RTX repeat-like sequence and 3 large repeat sequences that are partially similar to the outer membrane protein found in several prokaryotes. Recombinant PnxIIIA protein (rPnxIIIA) was cytotoxic toward J774A.1 mouse macrophage cells, whereas cytotoxicity was attenuated by the addition of anti-CD11a monoclonal antibody. rPnxIIIA could bind to extracellular matrices (ECMs) and cause hemagglutination of sheep erythrocytes. Binding was dependent on the 3 large repeat sequences in PnxIIIA. Protein interaction analyses indicated that PnxIIIA is mainly localized in the outer membrane of <it>P. pneumotropica </it>ATCC 35149 in a self-assembled oligomeric form. PnxIIIA is less cytotoxic to J774A.1 cells than PnxIA and PnxIIA.</p> <p>Conclusions</p> <p>The results implicate that PnxIIIA is located on the cell surface and participates in adhesion to ECMs and enhanced hemagglutination in the rodent pathogen <it>P. pneumotropica</it>.</p

Prevalence and analysis of Pseudomonas aeruginosa in chinchillas

<p>Abstract</p> <p>Background</p> <p>Chinchillas (<it>Chinchilla laniger</it>) are popular as pets and are often used as laboratory animals for various studies. <it>Pseudomonas aeruginosa </it>is a major infectious agent that causes otitis media, pneumonia, septicaemia enteritis, and sudden death in chinchillas. This bacterium is also a leading cause of nosocomial infections in humans. To prevent propagation of <it>P. aeruginosa </it>infection among humans and animals, detailed characteristics of the isolates, including antibiotic susceptibility and genetic features, are needed. In this study, we surveyed <it>P. aeruginosa </it>distribution in chinchillas bred as pets or laboratory animals. We also characterized the isolates from these chinchillas by testing for antibiotic susceptibility and by gene analysis.</p> <p>Results</p> <p><it>P. aeruginosa </it>was isolated from 41.8% of the 67 chinchillas included in the study. Slide agglutination and pulsed-field gel electrophoresis discriminated 5 serotypes and 7 unique patterns, respectively. For the antibiotic susceptibility test, 40.9% of isolates were susceptible to gentamicin, 77.3% to ciprofloxacin, 77.3% to imipenem, and 72.7% to ceftazidime. DNA analyses confirmed that none of the isolates contained the gene encoding extended-spectrum β-lactamases; however, 2 of the total 23 isolates were found to have a gene similar to the <it>pilL </it>gene that has been identified in the pathogenicity island of a clinical isolate of <it>P. aeruginosa</it>.</p> <p>Conclusions</p> <p><it>P. aeruginosa </it>is widely spread in chinchillas, including strains with reduced susceptibility to the antibiotics and highly virulent strains. The periodic monitoring should be performed to help prevent the propagation of this pathogen and reduce the risk of infection from chinchillas to humans.</p

Assessment of energy expenditure using doubly labeled water, physical activity by accelerometer and reported dietary intake in Japanese men with type 2 diabetes: A preliminary study

The aim of the present study was to determine the total energy expenditure, physical activity and dietary intake of men with type 2 diabetes mellitus and control participants without type 2 diabetes mellitus who were matched for age and body mass index. The participants in the present study were 12 well‐controlled type 2 diabetes mellitus patients and 10 controls, aged 40–75 years, with a body mass index <30 kg/m2. Total energy expenditure under free‐living conditions was assessed using the doubly labeled water method, and physical activity was measured using a triaxial accelerometer. Dietary intake was assessed using a self‐recorded food intake diary during the measurement period. Participants were instructed to record their dietary intake over 3 days, including 2 weekdays. Total energy expenditure was not significantly different between the groups (P = 0.153), nor were energy (P = 0.969) or macronutrient intakes. In conclusion, when age and body mass index are matched, total energy expenditure and self‐reported energy intake are not significantly different between type 2 diabetes mellitus patients and healthy controls

Bleb formation in small unruptured intracranial aneurysm as a predictor of early rupture

Small unruptured aneurysms are thought to have a low risk of rupture, but the management of such lesions is still controversial. A 73-year-old man with a small anterior communication artery aneurysm, 4 mm in diameter, while on follow-up, developed an aneurysmal subarachnoid hemorrhage 2 weeks after the detection of a newly emerged bleb on the surface of the aneurysm. In conclusion, the formation of a bleb should be considered as a warning sign of an impending rupture, and treatment should be provided even for patients with small aneurysms

Recurrent TMJ open lock

Patient: This report describes the case of a 51-year-old male patient who initially presented at age 23 with a habitual intermittent open lock (at > 35 mm) in the left temporomandibular joint (TMJ). The patient was able to manage this affliction through rapid-repetition jaw opening and closing. Tomography of the joint showed no irregular morphology, but intraoral examination revealed an occlusal interference at the mandibular left third molar during leftwards excursion. For this patient, alteration of lateral guidance using a palatal plate attached to the maxillary left canine precluded this intermittent open lock, but at 22 years of age, the open lock recurred and could not be relieved by the patient, who was unable to assume an occlusal position. Because conservative treatment was ineffective, a pumping manipulation technique was applied to reduce the open lock, after which the patient has maintained good jaw function. MRI taken before and after repositioning indicated that abrupt reduction of a displaced articular disk was the cause of the open lock, and that this articular disk was restored to its proper position during the manipulation. Discussion: Most TMJ open locks occur as anterior dislocation, where the mandibular head becomes trapped anterior to the articular eminences, causing excessive opening and difficulty closing. Our clinical findings from this patient indicate that open lock can occur through abrupt reduction of a displaced articular disk, particularly in patients with chronic internal derangement of the TMJ. Conclusion: TMJ open lock can occur following abrupt reduction of a displaced articular disk

CAR-T cell in vivo tracking method using PET scan with the reporter gene and new investigational tracer [18F] FHBG

Recent development of cancer immunotherapy, and CAR-T cell therapy in particular, attracts considerable attention among researchers. In antitumor immunity, cytotoxic T lymphocytes (CTLs) specifically recognize tumor-associated antigens (TAAs) in an MHC-class I-restricted manner and attack tumor cells. CTL activation and its clonal expansion require co-stimulatory receptors such as CD28, 4-1BB and OX40, and CTL stimulatory cytokine including IL-2 and IFNγ in addition to antigen recognition. However, tumor cells escape from the immune surveillance via decreased expression of MHC class 1, increased expression of co-inhibitory molecule, inhibitory cytokine production and accumulation of inhibitory immune cells during in vivo growth. CAR gene-designated extracellular binding domain consists of a target-specific antibody or a targeting ligand fused with the intracellular activating CD3zeta chain and the co-stimulatory molecule gene (1)

Differential activity of interferon-α8 promoter is regulated by Oct-1 and a SNP that dictates prognosis of glioma

We have previously reported that the single nucleotide polymorphism (SNP) rs12553612 in IFNA8 is associated with better overall survival of glioma patients with the AA-genotype compared with patients with the AC-genotype. As rs12553612 is located in the IFNA8 promoter, we hypothesized that the A-allele allows for an enhanced IFNA8 promoter activity compared with the C-allele. Reporter assays in the human monocyte derived THP-1 cell line demonstrated a superior promoter activity of the A-allele compared with the C-allele. Electrophoretic mobility shift assays (EMSA) further demonstrated that the A-genotype specifically binds to more nuclear proteins than the C-genotype, including the transcription factor Oct-1. Further, co-transfection of plasmids encoding Oct-1 and the reporter constructs revealed that Oct-1 enhanced the promoter activity with the A- but not the C-allele. Taken together, our data demonstrate that the A-allele in the rs12553612 SNP, which is associated with better glioma patient survival, allows for IFNA8 transcription by allowing for Oct-1 binding, which is absent in patients with C allele, and suggests a molecular mechanism of IFNA8 mediated immune-surveillance of glioma progression