Uptake of biodegradable poly(γ-glutamic acid) nanoparticles and antigen presentation by dendritic cells in vivo

AbstractPoly(γ-glutamic acid) (γ-PGA) nanoparticles (NPs) carrying antigens have been shown to induce potent antigen-specific immune responses. However, in vivo delivery of γ-PGA NPs to dendritic cells (DCs), a key regulator of immune responses, still remains unclear. In this study, γ-PGA NPs were examined for their uptake by DCs and subsequent migration from the skin to the regional lymph nodes (LNs) in mice. After subcutaneous injection of fluorescein 5-isothiocyanate (FITC)-labeled NPs or FITC-ovalbumin (OVA)-carrying NPs (FITC-OVA-NPs), DCs migrated from the skin to the LNs and maturated, resulting in the upregulation of the costimulatory molecules CD80 and CD86 and the chemokine receptor CCR7. However, the migrated DCs were not detected in the spleen. FITC-OVA-NPs were found to be taken up by skin-derived CD103+ DCs, and the processed antigen peptides were cross-presented by the major histocompatibility complex (MHC) class I molecule of DCs. Furthermore, significant activation of antigen-specific CD8+ T cells was observed in mice immunized with OVA-carrying NPs (OVA-NPs) but not with OVA alone or OVA with an aluminum adjuvant. The antigen-specific CD8+ T cells were induced within 7 days after immunization with OVA-NPs. Thus, γ-PGA NPs carrying various antigens may have great potential as an antigen-delivery system and vaccine adjuvant in vivo

CSC with and without steroids

We investigated the rates of the use of steroids in Japanese central serous chorioretinopathy (CSC) cases and differences in the characteristics of CSC with and without steroids. A total of 538 eyes of 477 patients diagnosed with CSC, with 3 months or more of follow-up between April 2013 and June 2017 at 8 institutions. Patients with CSC with more than 3 months of follow-up were identified by OCT and fluorescein angiography at 8 institutions. Data collected included patient demographics, history of corticosteroid medication and smoking, spherical errors, findings of angiography, subfoveal choroidal thickness, and changes through the follow-up period. Differences in these findings were analyzed in cases with and without corticosteroid treatment. Among the 477 patients (344 men,133 women), 74 (15.5%) (39 men, 35 women) underwent current or prior steroid treatment. Cases with steroids were higher age (p = 0.0403) and showed no male prevalence, more bilateral involvement (p < 0.0001), and the affected eyes had multiple pigment epithelial detachment (p <0.0001), more fluorescein leakage sites (p < 0.0001), greater choroidal thickness (p = 0.0287) and a higher recurrence rate (p = 0.0412). Steroids can cause severer CSC through an effect on choroidal vessels and an impairment of retinal pigment epithelium

光異性化反応によるスリンダクの生態毒性の増強

　近年、世界各地の河川や湖沼等の水環境中から医薬品を検出した事例が数多く報告されている。それに伴い、医薬品の生態系への影響や環境中での挙動が注目されているが、多くの医薬品において、知見に乏しいのが現状である。本研究では、解熱鎮痛剤であるスリンダクについて、水生生物に対する影響が紫外線照射により変化するかについて、ISO11348-1に準拠した生態毒性試験により検討した。すなわち、スリンダク、紫外線（主波長302 nm）を照射したスリンダクおよびスリンダク光分解物の水溶液を海洋発光細菌V. fischeriに暴露し、発光の減少量から毒性の大小を評価した。その結果、紫外線を照射することでスリンダクの毒性は有意に増強し、それには生成した光分解物であるスリンダクのtrans体が大きく寄与することが明らかとなった。今回の結果より、医薬品の環境リスクアセスメントを行う上で、親化合物だけでなくその環境下での分解物についても評価が必要であることが示唆された

Efficacy of gilteritinib in comparison with alectinib for the treatment of ALK-rearranged non-small cell lung cancer

Gilteritinib is a multitarget tyrosine kinase inhibitor (TKI), approved for the treatment of FLT3-mutant acute myeloid leukemia, with a broad range of activity against several tyrosine kinases including anaplastic lymphoma kinase (ALK). This study investigated the efficacy of gilteritinib against ALK-rearranged non-small cell lung cancers (NSCLC). To this end, we assessed the effects of gilteritinib on cell proliferation, apoptosis, and acquired resistance responses in several ALK-rearranged NSCLC cell lines and mouse xenograft tumor models and compared its efficacy to alectinib, a standard ALK inhibitor. Gilteritinib was significantly more potent than alectinib, as it inhibited cell proliferation at a lower dose, with complete attenuation of growth observed in several ALK-rearranged NSCLC cell lines and no development of drug tolerance. Immunoblotting showed that gilteritinib strongly suppressed phosphorylated ALK and its downstream effectors, as well as mesenchymal-epithelial transition factor (MET) signaling. By comparison, MET signaling was enhanced in alectinib-treated cells. Furthermore, gilteritinib was found to more effectively abolish growth of ALK-rearranged NSCLC xenograft tumors, many of which completely receded. Interleukin-15 (IL-15) mRNA levels were elevated in gilteritinib-treated cells, together with a concomitant increase in the infiltration of tumors by natural killer (NK) cells, as assessed by immunohistochemistry. This suggests that IL-15 production along with NK cell infiltration may constitute components of the gilteritinib-mediated antitumor responses in ALK-rearranged NSCLCs. In conclusion, gilteritinib demonstrated significantly improved antitumor efficacy compared with alectinib against ALK-rearranged NSCLC cells, which can warrant its candidacy for use in anticancer regimens, after further examination in clinical trial settings

Down-regulation of CD5 expression on activated CD8+ T cells in familial hemophagocytic lymphohistiocytosis with perforin gene mutations

Hemophagocytic lymphohistiocytosis (HLH) is characterized by uncontrolled activation of T cells and macrophages with overproduction of cytokines. Familial HLH type 2 (FHL2) is the most common form of primary HLH and is caused by mutations in PRF1. We have recently described a significant increase in the subpopulation of CD8+ T cells with clonal expansion and CD5 down-regulation in Epstein-Barr virus associated-HLH, which represented a valuable tool for its diagnosis. However, this unusual phenotype of CD8+ T cells has not been investigated fully in patients with FHL2. We performed immunophenotypic analysis of peripheral blood and measured serum pro-inflammatory cytokines in five patients with FHL2. All patients showed significantly increased subpopulations of activated CD8+ T cells with down-regulation of CD5, which were negligible among normal controls. Analysis of T-cell receptor Vβ repertoire suggested the reactive and oligoclonal expansion of these cells. The proportion of the subset declined after successful treatment concomitant with reduction in the serum levels of cytokines in all patients except one who continued to have a high proportion of the subset and died. These findings suggest that down-regulation of CD5 on activated CD8+ T cells may serve as a useful marker of dysregulated T cell activation and proliferation in FHL2. © 2013 American Society for Histocompatibility and Immunogenetics

コアシェル型逆相HPLCカラムを用いたコルチコステロイド類の迅速一斉分離での選択性改善と外用剤の定量法開発

　This paper describes separation of eight corticosteroids by the reversed-phase HPLC with core-shell type columns, which have a core part and a superficial porous silica part, leading to high performance (high theoretical plate number) and relatively low column pressure drops. First typical C18 and C28 columns having 2.6 μm particle size and a linear carbon chain structure were employed with the mobile phase containing acetonitrile (ACN) or methanol (MeOH) as an organic solvent. Among eight corticosteroids, cortisone and hydrocortisone,those only differ in C=O and C-OH at position 11, were co-eluted in use of ACN, on the other hand, triamcinolone acetonide and fluosinolone acetonide, those only differ F residue at position 6, were co-eluted in use of MeOH. However, all eight corticosteroids were successfully separated within 8 min by the mobile phase with tetrahydrofuran (THF). It was found that THF is quite effective for the improvement of the selectivity in the separation of corticosteroids, as already shown in the separation between berberine and palmatine, and between codeine and dihydrocodeine. This can be explained by the solubility of THF (octadecane can be soluble in THF), leading to the straight structure of the carbon chain of C18. Next, other than the straight carbon chain type columns (C18, C28), perfluorophenyl or biphenyl type columns were employed for the separation of eight corticosteroids. In use of these planer type columns, successful separation of eight corticosteroids was obtained within 8 min in use of ACN. This means the phenyl moiety of the stationary phase is effective for the small difference in the corticosteroid structure. Finally assay methods of fluosinolone acetonide formulations were developed and applied for the determination of the active ingredient in the cream and the ointment. Fast assay was successful within 5 min