Receptor-mediated endocytosis of Neisseria gonorrhoeae into primary human urethral epithelial cells: the role of the asialoglycoprotein receptor

H A Harvey; M P Jennings; C A Campbell; R Williams; M A Apicella

doi:10.1046/j.1365-2958.2001.02666.x

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Receptor-mediated endocytosis of Neisseria gonorrhoeae into primary human urethral epithelial cells: the role of the asialoglycoprotein receptor

Journal article

Peer reviewed

Receptor-mediated endocytosis of Neisseria gonorrhoeae into primary human urethral epithelial cells: the role of the asialoglycoprotein receptor

H A Harvey, M P Jennings, C A Campbell, R Williams and M A Apicella

Molecular microbiology, Vol.42(3), pp.659-672

11/2001

DOI: 10.1046/j.1365-2958.2001.02666.x

PMID: 11722733

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Abstract

Urethral epithelial cells are invaded by Neisseria gonorrhoeae during gonococcal infection in men. To understand further the mechanisms of gonococcal entry into host cells, we used the primary human urethral epithelial cells (PHUECs) tissue culture system recently developed by our laboratory. These studies showed that human asialoglycoprotein receptor (ASGP-R) and the terminal lactosamine of lacto-N-neotetraose-expressing gonococcal lipooligosaccharide (LOS) play an important role in invasion of PHUECs. Microscopy studies showed that ASGP-R traffics to the cell surface after gonococcal challenge. Co-localization of ASGP-R with gonococci was observed. As ASGP-R-mediated endocytosis is clathrin dependent, clathrin localization in PHUECs was examined after infection. Infected PHUECs showed increased clathrin recruitment and co-localization of clathrin and gonococci. Preincubating PHUECs in 0.3 M sucrose or monodansylcadaverine (MDC), which both inhibit clathrin-coated pit formation, resulted in decreased invasion. N. gonorrhoeae strain 1291 produces a single LOS glycoform that terminates with Gal(beta1-4)GlcNac(beta1-3)Gal(beta1-4)Glc (lacto-N-neotetraose). Invasion assays showed that strain 1291 invades significantly more than four isogenic mutants expressing truncated LOS. Sialylation of strain 1291 LOS inhibited invasion significantly. Preincubation of PHUECs in asialofetuin (ASF), an ASGP-R ligand, significantly reduced invasion. A dose-response reduction in invasion was observed in PHUECs preincubated with increasing concentrations of NaOH-deacylated 1291 LOS. These studies indicated that an interaction between lacto-N-neotetraose-terminal LOS and ASGP-R allows gonococcal entry into PHUECs.

Carbohydrate Sequence

Gonorrhea - microbiology

Asialoglycoprotein Receptor

Humans

Cells, Cultured

Lipopolysaccharides - metabolism

Molecular Sequence Data

Receptors, Cell Surface - metabolism

Male

Endocytosis

Amino Sugars - metabolism

Epithelial Cells - microbiology

Lipopolysaccharides - chemistry

Neisseria gonorrhoeae - chemistry

Urothelium - microbiology

Neisseria gonorrhoeae - genetics

Neisseria gonorrhoeae - metabolism

Neisseria gonorrhoeae - pathogenicity

Urethra - microbiology

Details

Title: Subtitle: Receptor-mediated endocytosis of Neisseria gonorrhoeae into primary human urethral epithelial cells: the role of the asialoglycoprotein receptor
Creators: H A Harvey - Department of Microbiology, Bowen Science Building, 51 Newton Road, The University of Iowa, Iowa City, IA 52242, USA
M P Jennings
C A Campbell
R Williams
M A Apicella
Resource Type: Journal article
Publication Details: Molecular microbiology, Vol.42(3), pp.659-672
DOI: 10.1046/j.1365-2958.2001.02666.x
PMID: 11722733
NLM abbreviation: Mol Microbiol
ISSN: 0950-382X
eISSN: 1365-2958
Grant note: AI38515 / NIAID NIH HHS AI43924 / NIAID NIH HHS AI18384 / NIAID NIH HHS
Language: English
Date published: 11/2001
Academic Unit: Microbiology and Immunology; Cardiovascular Medicine; Internal Medicine
Record Identifier: 9984094716302771

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