ArticleMicrobes and infection
Quantification of bacterial internalization by host cells using a beta-lactamase reporter strain: Neisseria gonorrhoeae invasion into cervical epithelial cells requires bacterial viability.
Article in Microbes and infection. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed, 15 citations in OpenAlex.
- Article
- Article
- Quantifying intracellular Mycobacterium tuberculosis: An essential issue for in vitro assays.MicrobiologyOpen · 2018Review
- nagZ Triggers Gonococcal Biofilm Disassembly.Scientific reports · 2016Article
- Determination ofAntimicrobial agents and chemotherapy · 2013Article
- Single cell measurements of vacuolar rupture caused by intracellular pathogens.Journal of visualized experiments : JoVE · 2013Article
- Neisseria gonorrhoeae breaches the apical junction of polarized epithelial cells for transmigration by activating EGFR.Cellular microbiology · 2013Article
- Neisseria gonorrhoeae-induced transactivation of EGFR enhances gonococcal invasion.Cellular microbiology · 2011Article
- Anthrax toxin uptake by primary immune cells as determined with a lethal factor-beta-lactamase fusion protein.PloS one · 2009Article
Corrections and comments
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
Abstract
Neisseria gonorrhoeae can invade into cervical epithelial cells to overcome this host defense barrier. We developed a beta-lactamase reporter system that allowed us to quantify at the single cell level if a host cell internalized a viable or nonviable microorganism. We autodisplayed beta-lactamase on the surface of FA1090 [FA1090Phi(bla-iga')] and demonstrated by confocal fluorescence microscopy and flow cytometry that FA1090Phi(bla-iga') cleaved the beta-lactamase substrate CCF2-AM loaded into host cells only when gonococci were internalized by these host cells. While FA1090Phi(bla-iga') adhered to almost all ME180 cells, viable N. gonorrhoeae were internalized by only a subset of cells during infection. Nonviable gonococci adhered to, but were not internalized by ME180 cells, and failed to recruit F-actin to sites of adherent bacteria. Overall, we show that epithelial cell invasion is a dynamic process that requires viable N. gonorrhoeae. We demonstrate the advantages of the beta-lactamase reporter system over the gentamicin protection assay in quantifying bacterial invasion. The reporter system that we have developed can be adapted to studying the internalization of any bacterial species into any host cell.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.