Effects of the immunoglobulin A1 protease on Neisseria gonorrhoeae trafficking across polarized T84 epithelial monolayers.
We previously demonstrated that the Neisseria IgA1 protease cleaves LAMP1 (lysosome-associated membrane protein 1), a major integral membrane glycoprotein of lysosomes, thereby accelerating its degradation rate in infected A431 human epidermoid carcinoma cells and resulting in the alteration of lysosomes in these cells. In this study, we determined whether the IgA1 protease also affects the trafficking of Neisseria gonorrhoeae across polarized T84 epithelial monolayers. We report that N. gonorrhoeae infection of T84 monolayers, grown on a solid substrate or polarized on semiporous membranes, also results in IgA1 protease-mediated reduction of LAMP1. We demonstrate that iga mutants in two genetic backgrounds exited polarized T84 monolayers in fewer numbers than the corresponding wild-type strains. Finally, we present evidence that these mutants have a statistically significant and reproducible defect in their ability to traverse T84 monolayers. These results add to our previous data by showing that the IgA1 protease alters lysosomal content in polarized as well as unpolarized cells and by demonstrating a role for the protease in the traversal of epithelial barriers by N. gonorrhoeae.
Hopper
S
S
Department of Molecular Microbiology and Immunology, Oregon Health Sciences University, Portland, Oregon 97201-3098, USA. hoppers@ohsu.edu
Vasquez
B
B
Merz
A
A
Clary
S
S
Wilbur
J S
JS
So
M
M
D015703:Antigens, CD / Q000032:analysis; D016764:Cell Polarity; D004848:Epithelium / Q000382:microbiology; D006801:Humans; D052118:Lysosomal-Associated Membrane Protein 1; D051907:Lysosomal Membrane Proteins; D008562:Membrane Glycoproteins / Q000032:analysis; D009068:Movement; D009154:Mutation; D009344:Neisseria gonorrhoeae / Q000502:physiology*; D012697:Serine Endopeptidases / Q000502:physiology*; D014407:Tumor Cells, Cultured
D015703:Antigens, CD; D052118:Lysosomal-Associated Membrane Protein 1; D051907:Lysosomal Membrane Proteins; D008562:Membrane Glycoproteins; D012697:Serine Endopeptidases; C021433:IgA-specific serine endopeptidase