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1.
Bio Protoc ; 8(5)2018 Mar 05.
Article in English | MEDLINE | ID: mdl-29623285

ABSTRACT

Here we describe a method to test bacterial adhesion to paraffin embedded tissue sections. This method allows examining binding of different bacterial strains, transfected with a fluorescent protein reporter plasmid to various tissues, to better understand different mechanisms such as colonization. This assay provides a more physiological context to bacterial binding, than would have been achieved using adhesion assays to cell lines. The sections can be imaged using fluorescent microscopy and adhesion of various bacterial strains can be quantified and tested, simultaneously.

2.
Cell Rep ; 20(1): 40-47, 2017 07 05.
Article in English | MEDLINE | ID: mdl-28683322

ABSTRACT

Urinary tract infection (UTI) is the most common type of bacterial infection in humans. Fifty percent of all women will experience at least one UTI in their lifetime, with uropathogenic Escherichia coli (UPEC) accounting for 80% of reported cases. UTI evokes a complex, well-timed immune response that is crucial for bacterial clearance. The majority of immune cells participating in the immune response are absent from the healthy bladder, and the mechanisms used to recruit them upon UTI are not fully understood. Here, we show that immediately after UPEC infection, bladder epithelial cells secrete stromal cell-derived factor 1 (SDF-1), initiating immune cell accumulation at the site of infection. SDF-1 blockade significantly reduced immune cell migration to the infected bladder, resulting in severe exacerbation of infection. We also show that FimH, the adhesin of type 1 fimbria, one of UPEC's virulence factors, is directly involved in the secretion of SDF-1 upon UTI.


Subject(s)
Chemokine CXCL12/immunology , Immunity, Innate , Urinary Tract Infections/immunology , Adhesins, Escherichia coli/immunology , Animals , Chemokine CXCL12/metabolism , Enteropathogenic Escherichia coli/immunology , Enteropathogenic Escherichia coli/pathogenicity , Female , Fimbriae Proteins/immunology , Killer Cells, Natural/immunology , Mice , Mice, Inbred C57BL , Neutrophils/immunology , T-Lymphocytes/immunology , Urinary Tract Infections/microbiology , Urothelium/immunology , Urothelium/metabolism
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