Vaccination with liposome-coupled glypican-3-derived epitope peptide stimulates cytotoxic T lymphocytes and inhibits GPC3-expressing tumor growth in mice

AbstractBecause therapeutic manipulation of immunity can induce tumor regression, anti-cancer immunotherapy is considered a promising treatment modality. We previously reported that glypican-3 (GPC3), an oncofetal antigen overexpressed in hepatocellular carcinoma (HCC), is a useful target for cytotoxic T lymphocyte (CTL)-mediated cancer immunotherapy, and we have performed clinical trials using the GPC3-derived peptide vaccine. Although vaccine-induced GPC3-peptide-specific CTLs were often tumor reactive in vitro and were correlated with overall survival, no complete response was observed. In the current study, we synthesized liposome-coupled GPC3-derived CTL epitope peptide (pGPC3-lipsome) and investigated its antitumor potential. Vaccination with pGPC3-liposome induced peptide-specific CTLs at a lower dose than conventional vaccine emulsified in incomplete Freund's adjuvant. Coupling of pGPC3 to liposomes was essential for effective priming of GPC3-specific CTLs. In addition, immunization with pGPC3-liposome inhibited GPC3-expressing tumor growth. Thus, vaccination with tumor-associated antigen-derived epitope peptides coupled to the surfaces of liposomes may be a novel therapeutic strategy for cancer

Porphyrin‐uptake in liposomes and living cells using an exchange method with cyclodextrin

The water‐solubilisation of porphyrin derivatives is very important for biological applications. Although liposomal drug carriers for porphyrin derivatives have shown significant promise in the field of medicinal chemistry (e.g., as sensitisers for photodynamic therapy), it is currently not possible to prepare lipid‐membrane‐incorporated tetraphenylporphyrin (TPP) with a high concentration of TPP using conventional methods. In this study, we have succeeded in preparing lipid-membrane‐incorporated TPP and zinc(II) tetraphenylporphyrin (ZnTPP) from the corresponding TPP or ZnTPP•cyclodextrin complex using the exchange method in lipid‐membranes composed of liposomes. Furthermore, the exchange method allowed for the incorporation of TPP or ZnTPP into the plasma membranes of HeLa cells. However, it was not possible to prepare lipid‐membrane‐incorporated porphyrin derivatives with polar and hydrophilic groups in the meso positions using this exchange reaction.Electronic supplementary information (ESI) available: Experimental procedures, 1H NMR spectra, DLS measurements, cryo-TEM images, phase contrast and fluorescence images. See DOI: 10.1039/c5ra24985This work was supported by JSPS KAKENHI a Grant‐in‐Aid for Scientific Research (B) (Grant No. 25288037) and a Grant‐in‐Aid for Young Scientists (A) (Grant No. 24681028)

Effects of Different Levels of Intraocular Stray Light on Kinetic Perimetry Findings

<div><p>Purpose</p><p>To evaluate the effect of different levels of intraocular stray light on kinetic perimetry findings.</p><p>Methods</p><p>Twenty-five eyes of 25 healthy young participants were examined by automated kinetic perimetry (Octopus 900) using Goldmann stimuli III4e, I4e, I3e, I2e, and I1e. Each stimulus was presented with a velocity of 3°/s at 24 meridians with 15° intervals. Four levels of intraocular stray light were induced using non-white opacity filter (WOF) filters and WOFs applied to the clear plastic eye covers of the participants. The visual acuity, pupil diameter, isopter area, and kinetic sensitivity of each meridian were analyzed for each WOF density.</p><p>Results</p><p>Visual acuity deteriorated with increasing WOF densities (p < 0.01). With a visual acuity of 0.1 LogMAR units, the isopter areas for III4e, I4e, I3e, I2e, and I1e decreased by -32.7 degree² (-0.2%), -255.7 degree² (-2.6%), -381.2 degree² (-6.2%), -314.8 degree² (-12.8%), and -59.2 degree² (-15.2%), respectively; kinetic sensitivity for those stimuli decreased by -0.1 degree (-0.1%), -0.8 degree (-1.4%), -1.6 degree (-3.7%), -2.7 degree (-9.7%), and -1.7 degree (-16.2%), respectively. The pupil diameter with each WOF density was not significantly different.</p><p>Conclusion</p><p>Kinetic perimetry measurements with a high-intensity stimulus (i.e., III4e) were unaffected by intraocular stray light. In contrast, measurements with the I4e, I3e, I2e, and I1e stimuli, especially I2e and I1e, were affected. Changes in the shape of the isopter resulting from opacity must be monitored, especially in cases of smaller and lower-intensity stimuli.</p></div

Measurable area and starting locations with a moving stimulus at each meridian.

<p>The Octopus 900 perimeter device can measure the area outlined by the dashed line. The starting locations with a moving stimulus at each meridian are depicted using the III4e stimulus as an example. The stimulus is presented randomly on each meridian from the extreme periphery of normal age-corrected kinetic sensitivity to the center. If the normal age-corrected kinetic sensitivity is outside the measurable area (dashed line), the starting location is set to the extreme end of the measurable area on the same meridian. The I4e, I3e, I2e, and I1e stimuli were also measured using the same method.</p

Sealed clear plastic eye cover with applied white opacity filters (WOFs).

<p>From left to right, non-WOF filter (Grade 0) and WOF densities of 0.8 (Grade 1), 0.4 (Grade 2), and 0.1 (Grade 3) are presented.</p

Demographic data and ocular characteristics of the participants.

<p>Demographic data and ocular characteristics of the participants.</p

Deterioration in the isopter area per 0.1 LogMAR unit at each stimulus.

<p>Colored dots indicate deterioration in the isopter area per 0.1 LogMAR unit for each participant and the dotted lines indicate approximate straight lines.</p

Kinetic sensitivity at each meridian of each stimulus under Grades 0 to 3.

<p>Each plot was expressed as an average value for all participants. The mean visual acuity (logarithmic minimum angle of resolution, LogMAR) and mean kinetic sensitivity measured for each stimulus are shown in the lower middle and right, respectively. The symbols * and ** indicate that the kinetic sensitivity significantly decreased with p < 0.05 and 0.01 compared to the baseline Grade 0, respectively.</p

Analyses of non-benzodiazepine-induced adverse events and prognosis in elderly patients based on the Japanese adverse drug event report database

Abstract Background The contents of the guidelines for the use of non-benzodiazepines (Z-drugs) differ slightly between THE JAPANESE SOCIETY OF SLEEP RESEARCH and THE JAPAN GERIATRIC SOCIETY, and the recommended directions are conflicting. Therefore, we analyzed the use of the Japanese Adverse Drug Event Report database (JADER) for identifying adverse events (AEs) caused by Z-drugs and clarifying their occurrence trend and prognosis. Methods The signal value for comparison was calculated by using the proportional reporting ratio (PRR) and chi-squared test (χ2) results of data of elderly and non-elderly patients. Among AEs for which signals were detected in the elderly, we determined that those with lower signal values for non-elderly patients that were half the signal value of the elderly should be used with particular caution in the elderly. We also compared the prognoses. Results The AEs with > 1 risk ratio (RR) in elderly and non-elderly patients were regarded as those that should be noted in the prognosis of AEs in elderly patients. Furthermore, 28 AEs were detected in elderly patients’ signals. In this study, in addition to movement disorders such as “falls” and “bone fractures,” identified by two academic societies, signal characteristics of the elderly were obtained for psychiatric disorders and eye disorders. Conclusions There was no difference in prognosis, but these disorders could reduce the quality of life of patients. Therefore, we consider that in prescribing appropriate drug therapy for insomnia, attention should be paid to the occurrence of the AEs caused by the Z-drugs revealed by this study and the guidelines

Deletion of Hypoxia-Inducible Factor-1α in Adipocytes Enhances Glucagon-Like Peptide-1 Secretion and Reduces Adipose Tissue Inflammation

<div><p>It is known that obese adipose tissues are hypoxic and express hypoxia-inducible factor (HIF)-1α. Although some studies have shown that the expression of HIF-1α in adipocytes induces glucose intolerance, the mechanisms are still not clear. In this study, we examined its effects on the development of type 2 diabetes by using adipocyte-specific HIF-1α knockout (ahKO) mice. ahKO mice showed improved glucose tolerance compared with wild type (WT) mice. Macrophage infiltration and mRNA levels of monocyte chemotactic protein-1 (MCP-1) and tumor necrosis factor α (TNFα) were decreased in the epididymal adipose tissues of high fat diet induced obese ahKO mice. The results indicated that the obesity-induced adipose tissue inflammation was suppressed in ahKO mice. In addition, in the ahKO mice, serum insulin levels were increased under the free-feeding but not the fasting condition, indicating that postprandial insulin secretion was enhanced. Serum glucagon-like peptide-1 (GLP-1) levels were also increased in the ahKO mice. Interestingly, adiponectin, whose serum levels were increased in the obese ahKO mice compared with the obese WT mice, stimulated GLP-1 secretion from cultured intestinal L cells. Therefore, insulin secretion may have been enhanced through the adiponectin-GLP-1 pathway in the ahKO mice. Our results suggest that the deletion of HIF-1α in adipocytes improves glucose tolerance by enhancing insulin secretion through the GLP-1 pathway and by reducing macrophage infiltration and inflammation in adipose tissue.</p></div