iPSC-derived hypoimmunogenic tissue resident memory T cells mediate robust anti-tumor activity against cervical cancer.

Functionally rejuvenated human papilloma virus-specific cytotoxic T lymphocytes (HPV-rejTs) generated from induced pluripotent stem cells robustly suppress cervical cancer. However, autologous rejT generation is time consuming, leading to difficulty in treating patients with advanced cancer. Although use of allogeneic HPV-rejTs can obviate this, the major obstacle is rejection by the patient immune system. To overcome this, we develop HLA-A24&-E dual integrated HPV-rejTs after erasing HLA class I antigens. These rejTs effectively suppress recipient immune rejection while maintaining more robust cytotoxicity than original cytotoxic T lymphocytes. Single-cell RNA sequencing performed to gain deeper insights reveal that HPV-rejTs are highly enriched with tissue resident memory T cells, which enhance cytotoxicity against cervical cancer through TGFßR signaling, with increased CD103 expression. Genes associated with the immunological synapse also are upregulated, suggesting that these features promote stronger activation of T cell receptor (TCR) and increased TCR-mediated target cell death. We believe that our work will contribute to feasible "off-the-shelf" T cell therapy with robust anti-cervical cancer effects.

Sustainable Tumor-Suppressive Effect of iPSC-Derived Rejuvenated T Cells Targeting Cervical Cancers.

Immunotherapy utilizing induced pluripotent stem cell (iPSC) technology has great potential. Functionally rejuvenated cytotoxic T lymphocytes (CTLs) can survive long-term as young memory T cells in vivo, with continuous tumor eradication. Banking of iPSCs as an unlimited "off-the-shelf" source of therapeutic T cells may be feasible. To generate safer iPSCs, we reprogrammed human papilloma virus type 16 (HPV16) E6-specific CTLs by Sendai virus vector without cotransduction of SV40 large T antigen. The iPSCs efficiently differentiated into HPV16-specific rejuvenated CTLs that demonstrated robust cytotoxicity against cervical cancer. The tumor-suppressive effect of rejuvenated CTLs was stronger and more persistent than that of original peripheral blood CTLs. These rejuvenated HPV16-specific CTLs provide a sustained tumor-suppressive effect even for epithelial cancers and constitute promising immunotherapy for cervical cancer.

An improved method for isolation of epithelial and stromal cells from the human endometrium.

We aimed to improve the efficiency of isolating endometrial epithelial and stromal cells (EMECs and EMSCs) from the human endometrium. We revealed by immunohistochemical staining that the large tissue fragments remaining after collagenase treatment, which are usually discarded after the first filtration in the conventional protocol, consisted of glandular epithelial and stromal cells. Therefore, we established protease treatment and cell suspension conditions to dissociate single cells from the tissue fragments and isolated epithelial (EPCAM-positive) and stromal (CD13-positive) cells by fluorescence-activated cell sorting. Four independent experiments showed that, on average, 1.2 × 10(6) of EMECs and 2.8 × 10(6) EMSCs were isolated from one hysterectomy specimen. We confirmed that the isolated cells presented transcriptomic features highly similar to those of epithelial and stromal cells obtained by the conventional method. Our improved protocol facilitates future studies to better understand the molecular mechanisms underlying the dynamic changes of the endometrium during the menstrual cycle.

A Pilot Study on the Efficacy of Continuous Positive Airway Pressure on the Manifestations of Ménière's Disease in Patients with Concomitant Obstructive Sleep Apnea Syndrome.

OBJECTIVES: To evaluate the effect of continuous positive airway pressure (CPAP) therapy on Ménière's disease patients with concomitant obstructive sleep apnea syndrome (OSAS), since recent reports suggest OSAS may cause dysfunction of the vestibular system. STUDY DESIGN: Prospective study using CPAP administered to patients diagnosed with "Definite Ménière's disease" according to the guidelines of the American Academy of Otolaryngology--Head and Neck Surgery and combined with OSAS. SETTING: University hospital. METHODS: Twenty consecutive patients, 14 male and 6 female with active, unilateral, cochleovestibular Ménière's disease refractory to medical management who also had concurrent OSAS as defined by International Classification of Sleep Disorders, Second Edition were selected to undergo solitary CPAP therapy. Audiometric testing, caloric testing, and DHI survey were conducted before and after CPAP therapy and compared to assess effectiveness of CPAP therapy as utilized for treatment of Ménière's disease. RESULTS: Although caloric testing did not show significant difference, audiometric testing and results of dizziness handicap inventory were significantly improved (p < 0.05) after CPAP therapy only, without standard treatment for Ménière's disease. CONCLUSION: Recent reports have suggested that OSAS may cause dysfunction of the vestibular system. We investigated whether standard therapy for OSAS would be of benefit in the management of vertigo and hearing loss in Ménière's disease patients. Our study cohort demonstrated significant improvement in both DHI and audiometric testing following solitary CPAP therapy for OSAS. Solitary CPAP therapy may become a new effective treatment strategy for Ménière's disease patients with OSAS, not just only for control of dizziness and vertigo but also for potential benefit of hearing.

[Clinical Analyses of 115 Patients with Peritonsillar Abscess].

The clinical data of 115 patients with peritonsillar abscess (98 men and 17 women) treated between May 2011 and March 2014 were analyzed. We examined 9 items; the age, sex, affected side, duration of hospitalization, method of drainage, smoking history, history of diabetes, antibacterial drugs used, and the isolated bacteria. The disease predominantly affected males in their 30s (27.8% of all the patients). The median duration of hospitalization was 7 days. In regard to the affected side, the right side was affected in 52%, the left side in 44%, and both sides in 4%. The method of drainage used was incision in 63%, and puncture in 37%. In regard to the personal and past medical history, 51% of patients had a history of smoking and 3.5% had a history of diabetes. ABPC/SBT was used as the single-agent antibacterial drug in 75% of cases. The most commonly isolated aerobic bacteria were α-hemolytic streptococci, and the most commonly isolated anaerobic bacteria were Prevotella. The duration of hospitalization showed no significant correlation with the smoking history, drainage method or the antibiotic treatment used (ABPC/SBT single-agent or multiple drug use). On the other hand, the duration of hospitalization was significantly longer in the more than ≥65 years' age group than in the <65 years' age group. Therefore, especially careful interventions for prevention and treatment of peritonsillar abscess are required in the elderly. In relation to antibiotic selection, it may be reasonable to expect sufficient effect with the use of ABPC/SBT as a single agent, as this antibiotic has a broad antibacterial spectrum covering aerobic, anaerobic and drug-resistant bacteria.

A temporal window for estimating surface brightness in the Craik-O'Brien-Cornsweet effect.

The central edge of an opposing pair of luminance gradients (COC edge) makes adjoining regions with identical luminance appear to be different. This brightness illusion, called the Craik-O'Brien-Cornsweet effect (COCe), can be explained by low-level spatial filtering mechanisms (Dakin and Bex, 2003). Also, the COCe is greatly reduced when the stimulus lacks a frame element surrounding the COC edge (Purves et al., 1999). This indicates that the COCe can be modulated by extra contextual cues that are related to ideas about lighting priors. In this study, we examined whether processing for contextual modulation could be independent of the main COCe processing mediated by the filtering mechanism. We displayed the COC edge and frame element at physically different times. Then, while varying the onset asynchrony between them and changing the luminance contrast of the frame element, we measured the size of the COCe. We found that the COCe was observed in the temporal range of around 600-800 ms centered at the 0 ms (from around -400 to 400 ms in stimulus onset asynchrony), which was much larger than the range of typical visual persistency. More importantly, this temporal range did not change significantly regardless of differences in the luminance contrast of the frame element (5-100%), in the durations of COC edge and/or the frame element (50 or 200 ms), in the display condition (interocular or binocular), and in the type of lines constituting the frame element (solid or illusory lines). Results suggest that the visual system can bind the COC edge and frame element with a temporal window of ~1 s to estimate surface brightness. Information from the basic filtering mechanism and information of contextual cue are separately processed and are linked afterwards.

[A case of blunt common carotid artery injury following minor trauma].

Cervical major blood vessel injuries often produce acute ingravescence of the circulatory dynamics. Therefore, if immediate treatment is not given, fatal complications can occur, resulting in death. Common carotid artery (CCA) injuries in particular are often associated with fatal outcomes. Moreover, most CCA injuries with hemorrhage producing hematoma are the result penetrating trauma, and there are few reports of blunt injuries. We report herein on a case of blunt CCA injury producing acute hematoma due to minor trauma. A 35-year-old man who was gently punched on his jaw when he was training with his child visited emergency room in our hospital complaining of swelling and pain of his neck soon after that. When we examined his neck, the larynx was displaced to the left by right neck swelling. Flexible transnasal laryngoscopic examination revealed constriction of the suffocating airway by a hematoma, so an emergency tracheotomy was performed. Enhanced CT of the neck showed active bleeding, so emergency surgical removal of the hematoma and hemostasis was carried out. We found a laceration (approximately 2 cm) of the CCA, and arrested hemorrhage with sutures. No postoperative neurologic deficit occurred. His postoperative course was good, and discharged 22 days after the operation.

Versican is upregulated in circulating monocytes in patients with systemic sclerosis and amplifies a CCL2-mediated pathogenic loop.

INTRODUCTION: Altered phenotypes of circulating monocytes of patients with systemic sclerosis (SSc) have been reported, but the role of these alterations in the pathogenesis of SSc remains unclear. This study was undertaken to identify molecules that are preferentially expressed by SSc monocytes, and to investigate the roles of these molecules in the pathogenic process of SSc. METHODS: We analyzed circulating CD14⁺ monocytes isolated from 36 patients with SSc and 32 healthy control subjects. The monocytes' gene expression profiles were assessed by Oligo GEArray (SABiosciences, Frederic, MA, USA) and semiquantitative or quantitative PCR; their protein expression was evaluated in culture supernatants of unstimulated monocytes by immunoblotting or ELISA, and by immunocytostaining. Monocyte chemoattractant activity of CCL2 was assessed in a TransWell system (Corning Incorporated, Corning, NY, USA) in the presence or absence of chondroitin sulfate (CS). RESULTS: A step-wise approach to profiling gene expression identified that versican and CCL2 were upregulated in SSc monocytes. Subsequent analysis of proteins expressed in monocyte culture supernatants confirmed enhanced production of versican and CCL2 in SSc monocytes compared with control monocytes. CCL2 bound to CS chains of versican and colocalized with versican in the monocytes' Golgi apparatus. Finally, CCL2 had a greater ability to mediate monocyte migration when bound to CS chains, because this binding provided efficient formation of CCL2 gradients and protection from protease attack. CONCLUSION: Circulating monocytes with elevated versican and CCL2 levels may contribute to the fibrotic process in a subset of SSc patients by amplifying a positive feedback loop consisting of versican, CCL2, and the influx of monocytes.

Awareness of Central Luminance Edge is Crucial for the Craik-O'Brien-Cornsweet Effect.

The Craik-O'Brien-Cornsweet (COC) effect demonstrates that perceived lightness depends not only on the retinal input at corresponding visual areas but also on distal retinal inputs. In the COC effect, the central edge of an opposing pair of luminance gradients (COC edge) makes adjoining regions with identical luminance appear to be different. To investigate the underlying mechanisms of the effect, we examined whether the subjective awareness of the COC edge is necessary for the generation of the effect. We manipulated the visibility of the COC edge using visual backward masking and continuous flash suppression while monitoring subjective reports regarding online percepts and aftereffects of adaptation. Psychophysical results showed that the online percept of the COC effect nearly vanishes in conditions where the COC edge is rendered invisible. On the other hand, the results of adaptation experiments showed that the COC edge is still processed at the early stage even under the perceptual suppression. These results suggest that processing of the COC edge at the early stage is not sufficient for generating the COC effect, and that subjective awareness of the COC edge is necessary.

A CD40-CD154 interaction in tissue fibrosis.

OBJECTIVE: To examine the role of an interaction between fibroblasts and mononuclear infiltrates through CD40-CD154 engagement in the development of tissue fibrosis. METHODS: Cultured dermal fibroblasts derived from healthy skin were induced to express CD40 by adenoviral gene transfer, stimulated with soluble CD154, and evaluated for proliferation, gene expression, and protein expression in vitro. The skin of mice with bleomycin-induced skin sclerosis, a model for systemic sclerosis (SSc), was assessed for CD40 and CD154 expression, in vivo fibroblast proliferation, and the expression of specific genes. The effects of an anti-CD154 monoclonal antibody on bleomycin-induced skin sclerosis were also examined. RESULTS: Upon stimulation with soluble CD154, cultured fibroblasts induced to express CD40 by adenoviral gene transfer proliferated and showed up-regulation of the genes for intercellular adhesion molecule 1, interleukin-6 (IL-6), IL-8, monocyte chemoattractant protein 1 (MCP-1), and RANTES, as well as up-regulation of their proteins. In the skin from bleomycin-treated mice, dermal fibroblasts expressed CD40, and mast cells and CD4+ T cells expressed CD154. Electron microscopic analysis revealed fibroblasts attached to mast cells and T cells with primitive contacts. The proliferation of fibroblasts and the up-regulated MCP-1 gene expression preceded thickening of the dermis. Finally, the anti-CD154 antibody inhibited the bleomycin-induced skin sclerosis by suppressing fibroblast proliferation and down-regulating MCP-1 expression. CONCLUSION: The interaction between fibroblasts and mast cells or T cells through CD40-CD154 signaling is critical for fibroblast activation early in the course of fibrosis. Blockade of the CD40-CD154 signal may be a novel therapeutic strategy for human fibrotic diseases, such as SSc.