Imaging Flow Cytometry Analysis of CEACAM Binding to Opa-Expressing Neisseria gonorrhoeae

Lacie M. Werner, Allison Palmer, Asya Smirnov, Meagan Belcher Dufrisne, Linda Columbus, Alison K. Criss

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

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.

Original languageEnglish
Pages (from-to)1081-1089
Number of pages9
JournalCytometry Part A
Volume97
Issue number10
DOIs
StatePublished - Oct 1 2020

Keywords

  • Neisseria gonorrhoeae
  • Opa protein
  • carcinoembryonic antigen-related cell adhesion molecule (CEACAM)
  • imaging flow cytometry
  • receptor-ligand binding

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