title: Imaging of Cell-Cell Communication in a Vertical Orientation Reveals High-Resolution Structure of Immunological Synapse and Novel PD-1 Dynamics.
abstract: The immunological synapse (IS) is one of the most pivotal communication strategies in immune cells. Understanding the molecular basis of the IS provides critical information regarding how immune cells mount an effective immune response. Fluorescence microscopy provides a fundamental tool to study the IS. However, current imaging techniques for studying the IS cannot sufficiently achieve high resolution in real cell-cell conjugates. In this study, we present a new device that allows for high-resolution imaging of the IS with conventional confocal microscopy in a high-throughput manner. Combining micropits and single-cell trap arrays, we have developed a new microfluidic platform that allows visualization of the IS in vertically "stacked" cells. Using this vertical cell pairing (VCP) system, we investigated the dynamics of the inhibitory synapse mediated by an inhibitory receptor, programed death protein-1, and the cytotoxic synapse at the single-cell level. In addition to the technique innovation, we have demonstrated novel biological findings by this VCP device, including novel distribution of F-actin and cytolytic granules at the IS, programed death protein-1 microclusters at the NK IS, and kinetics of cytotoxicity. We propose that this high-throughput, cost-effective, easy-to-use VCP system, along with conventional imaging techniques, can be used to address a number of significant biological questions in a variety of disciplines.
authors.affiliation: Department of Nanomedicine, Houston Methodist Research Institute, Houston, TX 77030; Center for Human Immunobiology, Texas Children's Hospital, Department of Pediatrics, Baylor College of Medicine, Houston, TX 77030;
authors.forename: Joon Hee
authors.initials: JH
authors.lastname: Jang
authors.affiliation: Center for Human Immunobiology, Texas Children's Hospital, Department of Pediatrics, Baylor College of Medicine, Houston, TX 77030; Department of Integrative Biology and Pharmacology, Graduate Program in Cell and Regulatory Biology, University of Texas Health Science Center at Houston, Houston, TX 77030;
authors.forename: Yu
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Center for Human Immunobiology, Texas Children's Hospital, Department of Pediatrics, Baylor College of Medicine, Houston, TX 77030;
authors.forename: Peilin
authors.initials: P
authors.lastname: Zheng
authors.affiliation: Department of Nanomedicine, Houston Methodist Research Institute, Houston, TX 77030;
authors.forename: Myeong Chan
authors.initials: MC
authors.lastname: Jo
authors.affiliation: Center for Human Immunobiology, Texas Children's Hospital, Department of Pediatrics, Baylor College of Medicine, Houston, TX 77030; Department of Pathology and Immunology, Baylor College of Medicine, Houston, TX 77030; and.
authors.forename: Grant
authors.initials: G
authors.lastname: Bertolet
authors.affiliation: Department of Integrative Biology and Pharmacology, Graduate Program in Cell and Regulatory Biology, University of Texas Health Science Center at Houston, Houston, TX 77030;
authors.forename: Michael Xi
authors.initials: MX
authors.lastname: Zhu
authors.affiliation: Department of Nanomedicine, Houston Methodist Research Institute, Houston, TX 77030; Department of Cell and Developmental Biology, Weill Cornell Medical College, New York, NY 10065 LQin@tmhs.org dongfang.liu@bcm.edu.
authors.forename: Lidong
authors.initials: L
authors.lastname: Qin
authors.affiliation: Center for Human Immunobiology, Texas Children's Hospital, Department of Pediatrics, Baylor College of Medicine, Houston, TX 77030; Department of Pathology and Immunology, Baylor College of Medicine, Houston, TX 77030; and LQin@tmhs.org dongfang.liu@bcm.edu.
authors.forename: Dongfang
authors.initials: D
authors.lastname: Liu
mesh_terms: D002450:Cell Communication / Q000276:immunology*; D045744:Cell Line, Tumor; D003602:Cytotoxicity, Immunologic / Q000276:immunology; D057809:HEK293 Cells; D006801:Humans; D054992:Immunological Synapses / Q000276:immunology* / Q000378:metabolism; D007694:Killer Cells, Natural / Q000276:immunology; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods; D061026:Programmed Cell Death 1 Receptor / Q000378:metabolism*
chemical_list: C105992:PDCD1 protein, human; D061026:Programmed Cell Death 1 Receptor