T cell migration in microchannels densely packed with T cells

T cells migrate diverse microenvironments of the body to mount antigen-specific immune responses. T cell activation, a key initial process for antigen-specific immune responses, occur in secondary lymphoid organs such as spleens and lymph nodes where high density of T cells migrates rapidly through the reticular networks formed by stromal cells. In vitro model system recapitulating key characteristics of secondary lymphoid organs, confined spaces densely packed with rapidly migrating cells, would be useful to investigate mechanisms ofT cell migration. In this study, we devised a method to fabricate microchannels densely packed with T cells. Microchannel arrays with fixed height (4 mu m) and length (1.5 mm) and various widths (15 similar to 80 mu m) were fabricated in between trapezoid-shaped reservoirs that facilitated T cell sedimentation near microchannel entries. Microchannel surface chemistry and filling time were optimized to achieve high packing density (0.89) of T cell filling within microchannels. Particle image velocimetry (PIV) analysis method was employed to extract velocity field of microchannels densely packed with T cells. Using velocity field information, various motility parameters were further evaluated to quantitatively assess the effects of microchannel width and media tonicity on T cell motility within cell dense microenvironments.Y

Turning behaviors of T cells climbing up ramp-like structures are regulated by myosin light chain kinase activity and lamellipodia formation

T cells navigate diverse microenvironments to perform immune responses. Micro-scale topographical structures within the tissues, which may inherently exist in normal tissues or may be formed by inflammation or injury, can influence T cell migration, but how T cell migration is affected by such topographical structures have not been investigated. In this study, we fabricated ramp-like structures with a 5 mu m height and various slopes, and observed T cells climbing up the ramp-like structures. T cells encountering the ramp-like structures exhibited MLC accumulation near head-tail junctions contacting the ramp-like structures, and made turns to the direction perpendicular to the ramp-like structures. Pharmacological study revealed that lamellipodia formation mediated by arp2/3 and contractility regulated by myosin light chain kinase (MLCK) were responsible for the intriguing turning behavior of T cells climbing the ramp-like structures. Arp2/3 or MLCK inhibition substantially reduced probability of T cells climbing sharp-edged ramp-like structures, indicating intriguing turning behavior of T cells mediated by lamellipodia formation and MLCK activity may be important for T cells to access inflamed or injured tissues with abrupt topographical changes.11Ysciescopu

Roles of endothelial A-type lamins in migration of T cells on and under endothelial layers

Stiff nuclei in cell-dense microenvironments may serve as distinct biomechanical cues for cell migration, but such a possibility has not been tested experimentally. As a first step addressing this question, we altered nuclear stiffness of endothelial cells (ECs) by reducing the expression of A-type lamins using siRNA, and investigated the migration of T cells on and under EC layers. While most T cells crawling on control EC layers avoided crossing over EC nuclei, a significantly higher fraction of T cells on EC layers with reduced expression of A-type lamins crossed over EC nuclei. This result suggests that stiff EC nuclei underlying T cells may serve as "duro-repulsive" cues to direct T cell migration toward less stiff EC cytoplasm. During subendothelial migration under EC layers with reduced expression of A-type lamins, T cells made prolonged contact and substantially deformed EC nuclei, resulting in reduced speed and directional persistence. This result suggests that EC nuclear stiffness promotes fast and directionally persistent subendothelial migration of T cells by allowing minimum interaction between T cells and EC nuclei.open11102sciescopu

The Effectiveness of Short Message Service to Assure the Preparation-to-Colonoscopy Interval before Bowel Preparation for Colonoscopy

Background/Aims. The preparation-to-colonoscopy (PC) interval is one of several important factors for the bowel preparation. Short message service (SMS) reminder from a cellular phone has been suggested to improve compliance in various medical situations. We evaluated the effectiveness of SMS reminders to assure the PC interval for colonoscopy. Methodology. This prospective randomized study was investigator blinded. In the No-SMS group, patients took the first 2 L polyethylene glycol (PEG) between 6 and 8 PM on the day before colonoscopy and the second 2 L PEG approximately 6 hours before the colonoscopy without SMS. In the SMS group, patients took first 2 L PEG in the same manner as the No-SMS group and the second 2 L PEG after receiving an SMS 6 hours before the colonoscopy. Results. The SMS group had a lower score than the No-SMS group, according to the Ottawa Bowel Preparation Scale (P<0.001). Multivariate logistic regression analysis showed that compliance with diet instructions (odds ratio (OR) 2.109; 95% confidence interval (CI), 1.11–3.99, P=0.022) and intervention using SMS ((OR) 2.329; 95% (CI), 1.34–4.02, P=0.002) were the independent significant factors for satisfactory bowel preparation. Conclusions. An SMS reminder to assure PC interval improved the bowel preparation quality for colonoscopy with bowel preparation

Differentially Expressed Potassium Channels Are Associated with Function of Human Effector Memory CD8+T cells

The voltage-gated potassium channel, Kv1.3, and the Ca2+-activated potassium channel, KCa3.1, regulate membrane potentials in T cells, thereby controlling T cell activation and cytokine production. However, little is known about the expression and function of potassium channels in human effector memory ( EM) CD8+ T cells that can be further divided into functionally distinct subsets based on the expression of the interleukin ( IL)-7 receptor alpha ( IL-7R alpha) chain. Herein, we investigated the functional expression and roles of Kv1.3 and KCa3.1 in EM CD8+ T cells that express high or low levels of the IL-7 receptor alpha chain ( IL-7R alpha(high) and IL-7R alpha(low), respectively). In contrast to the significant activity of Kv1.3 and KCa3.1 in IL-7Rahigh EM CD8+ T cells, IL-7Ralow EM CD8+ T cells showed lower expression of Kv1.3 and insignificant expression of KCa3.1. Kv1.3 was involved in the modulation of cell proliferation and IL-2 production, whereas KCa3.1 affected the motility of EM CD8+ T cells. The lower motility of IL-7Ralow EM CD8+ T cells was demonstrated using transendothelial migration and motility assays with intercellular adhesion molecule 1-and/or chemokine stromal cell-derived factor-1 alpha-coated surfaces. Consistent with the lower migration property, IL-7Ralow EM CD8+ T cells were found less frequently in human skin. Stimulating IL-7Ralow EM CD8+ T cells with IL-2 or IL-15 increased their motility and recovery of KCa3.1 activity. Our findings demonstrate that Kv1.3 and KCa3.1 are differentially involved in the functions of EM CD8+ T cells. The weak expression of potassium channels in IL-7Ralow EM CD8+ T cells can be revived by stimulation with IL-2 or IL-15, which restores the associated functions. This study suggests that IL-7Rahigh EM CD8+ T cells with functional potassium channels may serve as a reservoir for effector CD8+ T cells during peripheral inflammation.112Ysciescopu

Auto-Networking Technologies for IPv6 Mobile Ad Hoc Networks

Abstract. This paper presents auto-networking technologies for IPv6 mobile ad hoc networks. The auto-networking technologies consist of IPv6 unicast address autoconfiguration, IPv6 multicast address allocation, secure multicast DNS, and service discovery. These technologies are based on IPv6’s inherent autoconfiguration facility, which can provide ad hoc users with automatic networking in IPv6 ad hoc environment.

Multifunctional Microwell Arrays for Single Cell Level Functional Analysis of Lymphocytes

Functional analysis of lymphocytes is important for development of vaccines and diagnosis/treatment of various immune-related diseases. In this review, we describe multifunctional microwell arrays that enable functional analysis of lymphocytes at the single cell level. We first discuss key parameters for microwell array design. Then, we describe how different types of multifunctional microwell arrays were developed for various applications, including live cell imaging of lymphocyte activation, proliferation, and differentiation, and analyses of effector functions such as cytokine secretion and target cell lysis. Incorporation of novel surface chemistries and functional materials into microwell arrays for enhancing sensing capabilities will widen applications of this technology. Multifunctional microwell arrays will be a powerful tool for the development of novel therapeutics against immune-related diseases, in particular, for cancer immunotherapy.11Nsciescopu