ATP and its metabolite adenosine cooperatively upregulate the antigen-presenting molecules on dendritic cells leading to IFN-gamma production by T cells

Dendritic cells (DCs) present foreign antigens to T cells via the major histocompatibility complex (MHC), thereby inducing acquired immune responses. ATP accumulates at sites of inflammation or in tumor tissues, which triggers local inflammatory responses. However, it remains to be clarified how ATP modulates the functions of DCs. In this study, we investigated the effects of extracellular ATP on mouse bone marrow- derived dendritic cells (BMDCs) as well as the potential for subsequent T cell activation. We found that high concentrations of ATP (1 mM) upregulated the cell surface expression levels of MHC-I, MHC-II, and co-stimulatory molecules CD80 and CD86 but not those of co-inhibitory molecules PD-L1 and PD-L2 in BMDCs. Increased surface expression of MHC-I, MHC-II, CD80, and CD86 was inhibited by a pan-P2 receptor antagonist. In addition, the upregulation of MHC-I and MHC-II expression was inhibited by an adenosine P1 receptor antagonist and by inhibitors of CD39 and CD73, which metabolize ATP to adenosine. These results suggest that adenosine is required for the ATP-induced upregulation of MHC-I and MHC-II. In the mixed leukocyte reaction assay, ATP-stimulated BMDCs activated CD4 and CD8T cells and induced interferon-gamma (IFN-gamma) production by these T cells. Collectively, these results suggest that high concentrations of extracellular ATP upregulate the expression of antigenpresenting and co-stimulatory molecules but not that of coinhibitory molecules in BMDCs. Cooperative stimulation of ATP and its metabolite adenosine was required for the upregulation of MHC-I and MHC-II. These ATP-stimulated BMDCs induced the activation of IFN-gamma-producing T cells upon antigen presentation

High-depth spatial transcriptome analysis by photo-isolation chemistry

光照射を用いた超高解像度な遺伝子解析技術の開発に成功 --組織内に潜むがん細胞の病理診断などに応用可能--. 京都大学プレスリリース. 2021-07-27.In multicellular organisms, expression profiling in spatially defined regions is crucial to elucidate cell interactions and functions. Here, we establish a transcriptome profiling method coupled with photo-isolation chemistry (PIC) that allows the determination of expression profiles specifically from photo-irradiated regions of interest. PIC uses photo-caged oligodeoxynucleotides for in situ reverse transcription. PIC transcriptome analysis detects genes specifically expressed in small distinct areas of the mouse embryo. Photo-irradiation of single cells demonstrated that approximately 8, 000 genes were detected with 7 × 10⁴ unique read counts. Furthermore, PIC transcriptome analysis is applicable to the subcellular and subnuclear microstructures (stress granules and nuclear speckles, respectively), where hundreds of genes can be detected as being specifically localised. The spatial density of the read counts is higher than 100 per square micrometre. Thus, PIC enables high-depth transcriptome profiles to be determined from limited regions up to subcellular and subnuclear resolutions

A case of metastatic cancer with markedly elevated PSA level that was not detected by repeat prostate biopsy

BACKGROUND: Prostate-specific antigen (PSA) is a widely used specific tumor marker for prostate cancer. We experienced a case of metastatic prostate cancer that was difficult to detect by repeat prostate biopsy despite a markedly elevated serum PSA level. CASE PRESENTATION: A 64-year-old man was referred to our hospital with lumbar back pain and an elevated serum PSA level of 2036 ng/mL. Computed tomography, bone scintigraphy, and magnetic resonance imaging showed systemic lymph node and osteoblastic bone metastases. Digital rectal examination revealed a small, soft prostate without nodules. Ten-core transrectal prostate biopsy yielded negative results. Androgen deprivation therapy (ADT) was started because of the patient’s severe symptoms. Twelve-core repeat transrectal prostate biopsy performed 2 months later, and transurethral resection biopsy performed 5 months later, both yielded negative results. The patient refused further cancer screening because ADT effectively relieved his symptoms. His PSA level initially decreased to 4.8 ng/mL, but he developed castration-resistant prostate cancer 7 months after starting ADT. He died 21 months after the initial prostate biopsy from disseminated intravascular coagulation. CONCLUSION: CUP remains a considerable challenge in clinical oncology. Biopsies of metastatic lesions and multimodal approaches were helpful in this case

Mice Treated with an Antibiotic Cocktail Containing Metronidazole Show Attenuated Allergy Responses through the Induction of Regulatory T Cells in the Intestinal Mucosa

This study sought to determine whether antibiotic treatments affect the pathogenesis of allergic diseases. We demonstrated that mice treated with a mixture of four antibiotic agents （ampicillin, neomycine, metronidazole, and vancomycin） showed significantly decreased antigen-specific IgE production compared to control mice, whereas mice treated with the same cocktail without metronidazole showed no change in IgE production compared to controls. Regulatory T cell suppression induced by particular antibody combinations is a possible mechanism of this effect, with an increased frequency of regulatory T cells also observed in the mesenteric lymph nodes （MLN） of mice treated with the mixture of four antibiotics, but not in the second cocktail without metronidazole. Our antibiotic treatments also dramatically decreased the diversity of mouse intestinal microbiota compared to control mice, based on 16SrRNA analysis of the cecum fecal composition. In conclusion, mice treated with antibiotic mixtures containing metronidazole, but not those treated without metronidazole, showed attenuated allergy responses and increased induction of regulatory T cells in the mucosa

Alpha7 nicotinic ACh receptor mediated neuroprotective action by nicotine and GTS-21: An approach by the hippocampal organotypic slice cultures.

Nicotine, main constituent of tabaco, is known as a nicotinic acetylcholine receptor (nAChR) agonist and increases cognitive performance. 3-(2,4-dimethoxybenzylidene)-anabaseine (GTS-21) is derived from the marine worm toxin, anabaseine, and is alpha7-selective nAChR (D7-nAChR) agonist. Both nicotine and GTS-21 were expected as therapeutic agents of Alzheimer’s disease. Several studies showed that nicotine and GTS-21 protected neuron by activating nAChR, especially D7-nAChR. It has been reported that D7-nAChR has been shown to be an essential regulator of inflammation. The purpose of this study is to examine the neuroprotective and anti-inflammatory effects of nicotine and GTS-21 using organotypic hippocampal slice cultures. Kainic acid (KA, 5-50µM) induced concentration- and time-dependent neuronal cell death in the hippocampal organotypic slice cultures. The pretreatment with nicotine and GTS-21 tended to decrease in KA toxicity. In a CA3 area-specific analysis, pretreatment with nicotine resulted in significant inhibition of KA-induced neurotoxicity. The results suggest that nicotine may protect KA-induced neuronal cell death via D7-nAChR. We also examined anti-inflammatory effects of nicotine and GTS-21. Hippocampal slices were pretreated with nicotine or GTS-21, and then treated with lipopolysaccharide (LPS). LPS treatment induced concentration-dependent increases in TNFD and IL-1E gene expressions. LPS-induced TNFD gene expression, but not IL-1E was suppressed by GTS-21 pretreatment. These results suggest that D7-nAChR might be involved in the microglia activation towards a neuroprotective role by suppressing inflammatory cytokine

Operation strategy by target species on offshore purse seine fisheries in the western waters of Japan

東京海洋大学海洋科学部海洋生物資源学科東京海洋大学海洋科学部海洋生物資源学科山二漁業有限会社財団法人漁場油濁被害救済基金西日本魚市株式会社西日本魚市株式会

Seasonal patterns of fishing ground by target species on offshore purse seine fisheries in the western waters of Japan

東京海洋大学海洋科学部海洋生物資源学科東京海洋大学海洋科学部海洋生物資源学科山二漁業有限会社財団法人漁場油濁被害救済基金西日本魚市株式会社西日本魚市株式会

Heterogeneous circulating miRNA profiles of PBMAH

ObjectivePrimary bilateral macronodular adrenal hyperplasia (PBMAH), a rare cause of Cushing syndrome, is often diagnosed as a bilateral adrenal incidentaloma with subclinical cortisol production. Circulating microRNAs (miRNAs) are a characteristic of adrenocortical adenomas, but miRNA expression in PBMAH has not been investigated. We aimed to evaluate the circulating miRNA expression in patients with PBMAH and compare them with those in patients with non-functioning adrenocortical adenoma (NFA) and cortisol-producing adrenocortical adenoma (CPA).MethodsmiRNA profiling of plasma samples from four, five, and five patients with NFA, CPA, and PBMAH, respectively, was performed. Selected miRNA expressions were validated using quantitative RT-PCR.ResultsPBMAH samples showed distinct miRNA expression signatures on hierarchical clustering while NFA and CPA samples were separately clustered. PBMAH was distinguished from the adenoma group of NFA and CPA by 135 differentially expressed miRNAs. Hsa-miR-1180-3p, hsa-miR-4732-5p, and hsa-let-7b-5p were differentially expressed between PBMAH and adenoma (P = 0.019, 0.006, and 0.003, respectively). Furthermore, PBMAH could be classified into two subtypes based on miRNA profiling: subtype 1 with a similar profile to those of adenoma and subtype 2 with a distinct profile. Hsa-miR-631, hsa-miR-513b-5p, hsa-miR-6805-5p, and hsa-miR-548av-5p/548k were differentially expressed between PBMAH subtype 2 and adenoma (P = 0.027, 0.027, 0.027, and 1.53E-04, respectively), but not between PBMAH, as a whole, and adenoma.ConclusionCirculating miRNA signature was identified specific for PBMAH. The existence of subtype-based miRNA profiles may be associated with the pathophysiological heterogeneity of PBMAH