Imaging Flow Cytometry Analysis of CEACAM Binding to Opa‐Expressing Neisseria gonorrhoeae. Issue 10 (2nd June 2020)

• Record Type: Journal Article
• Authors: Werner, Lacie M.; Palmer, Allison; Smirnov, Asya; Belcher Dufrisne, Meagan; Columbus, Linda; Criss, Alison K.
• Is Part Of: Cytometry. Volume 97:Issue 10(2020)
• Page Start: 1081
• Page End: 1089
• Publication Date: 2020-06-02
• Subjects: imaging flow cytometry -- carcinoembryonic antigen‐related cell adhesion molecule (CEACAM) -- Neisseria gonorrhoeae -- Opa protein -- receptor‐ligand binding; Flow cytometry -- Periodicals; Imaging systems in biology -- Periodicals; Imaging systems in medicine -- Periodicals; Diagnostic imaging -- Periodicals; 571.605
• DOI: 10.1002/cyto.a.24037
• Languages: English
• ISSNs: 1552-4922

Abstract

Abstract: Human carcinoembryonic antigen‐related cell adhesion molecules (CEACAMs) are a family of receptors that mediate intercellular interactions. Pathogenic bacteria have ligands that bind CEACAMs on human cells. Neisseria gonorrhoeae (Gc) encodes numerous unique outer membrane opacity‐associated (Opa) proteins that are ligands for one or more CEACAMs. CEACAMs that are expressed on epithelial cells facilitate Gc colonization, while those expressed on neutrophils affect phagocytosis and consequent intracellular survival of Gc. Since Opa protein expression is phase‐variable, variations in receptor tropism affect how individual bacteria within a population interact with host cells. Here we report the development of a rapid, quantitative method for collecting and analyzing fluorescence intensity data from thousands of cells in a population using imaging flow cytometry to detect N‐CEACAM bound to the surface of Opa‐expressing Gc. We use this method to confirm previous findings regarding Opa‐CEACAM interactions and to examine the receptor–ligand interactions of Gc expressing other Opa proteins, as well as for other N‐CEACAM proteins. © 2020 International Society for Advancement of Cytometry Abstract : Here we describe an imaging flow cytometry method for analyzing the binding of the soluble N‐terminal domain of human carcinoembryonic antigen‐related cell adhesion molecules (CEACAMs) to Neisseria gonorrhoeae bacteria expressing one of several opacity (Opa) proteins. For submicron‐sized bacteria like N. gonorrhoeae, imaging flow cytometry is an advantage since the pixel intensities of particle images are used to distinguish intact bacteria from bacterial aggregates or debris, and allows thousands of bacteria in the population to be analyzed.