title: Quantification of natural killer cell polarization and visualization of synaptic granule externalization by imaging flow cytometry.
abstract: Defining immunological mechanisms underlying NK cell biology is crucial for the treatment and prevention of immune deficiency and malignancy. The limited availability of human biological specimens presents a challenge to the study of human immunobiology. The use of high throughput, multi-parametric assays will not only aid in the definition and diagnosis of complex human immune disorders affecting NK cell function but also advance NK cell biology through population-based assessment of molecular signaling. In an effort to garner the most information from limited numbers of human cells, we designed a quantitative method to study NK cell function using imaging flow cytometry (IFC), which combines multiparametric flow cytometry and fluorescence microscopy. Specifically, we developed IFC as a tool to measure polarization and secretion of lytic granules at the immunological synapse formed between an NK cell and a susceptible target. We have further validated our approach through quantitative comparison with high-resolution confocal microscopy. We show that IFC can be used as a quantitative, high throughput measure of NK cell biological function possessing greater dimensionality than standard flow cytometry.
authors.affiliation: Rice University, Houston, TX 77005, USA.|Center for Human Immunobiology, Texas Children's Hospital and Baylor College of Medicine, Houston, TX 77030, USA.
authors.forename: Dixita I
authors.initials: DI
authors.lastname: Viswanath
authors.affiliation: Center for Human Immunobiology, Texas Children's Hospital and Baylor College of Medicine, Houston, TX 77030, USA.
authors.forename: Emily M
authors.initials: EM
authors.lastname: Mace
authors.affiliation: Center for Human Immunobiology, Texas Children's Hospital and Baylor College of Medicine, Houston, TX 77030, USA.
authors.forename: Hsiang-Ting
authors.initials: HT
authors.lastname: Hsu
authors.affiliation: Rice University, Houston, TX 77005, USA.|Center for Human Immunobiology, Texas Children's Hospital and Baylor College of Medicine, Houston, TX 77030, USA.
authors.forename: Jordan S
authors.initials: JS
authors.lastname: Orange
mesh_terms: D015550:Cell Degranulation; D002460:Cell Line; D045744:Cell Line, Tumor; D005434:Flow Cytometry; D006801:Humans; D054992:Immunological Synapses; D007694:Killer Cells, Natural / Q000166:cytology* / Q000276:immunology*; D008856:Microscopy, Fluorescence
keywords: Degranulation; Flow cytometry; Imaging flow cytometry; Immunological synapse; Microscopy; Natural killer cell