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BMC Microbiol ; 16: 215, 2016 09 15.
Article in English | MEDLINE | ID: mdl-27633511

ABSTRACT

BACKGROUND: Pertussis or whooping cough is an acute respiratory illness caused by the Gram-negative pathogen Bordetella pertussis. Despite high vaccination coverage whooping cough is currently re-emerging in many developed countries. Although the causes of pertussis resurgence are matter of debate, emerging evidences suggest that acellular vaccines efficiently protect against the hallmark symptoms of pertussis disease but fail to prevent colonization. This presumably impacts on increased risk of bacterial transmission and consequent spread throughout the population. These evidences suggest that improved vaccines may be required for efficient bacterial clearance in the upper respiratory tract. Consequently, there is a need for novel bioassays to evaluate at pre-clinical or clinical level the impact of different vaccines on B. pertussis colonization. RESULTS: We developed a high-throughput bacterial adhesion inhibition (BAI) assay based on human respiratory cell lines and on live bacteria chemically conjugated to a fluorescent dye. Employing A549 cells as model, we evaluated the impact of antibodies elicited by acellular (aP) and whole cell (wP) vaccines on B. pertussis adhesion in vitro. Moreover, we settled the method also on polarized Calu-3 cells grown at air-liquid interface (ALI), showing that this assay can be extended to more complex cell models mimicking the airway epithelium. CONCLUSIONS: We proved that this method is a sensitive, rapid and reproducible system to evaluate the anti-adhesive properties of vaccine-induced antibodies and can be employed to assess improved pertussis vaccines.


Subject(s)
Adhesins, Bacterial/analysis , Bordetella pertussis/drug effects , Epithelial Cells/microbiology , High-Throughput Screening Assays/methods , Pertussis Vaccine/analysis , Respiratory System/microbiology , A549 Cells/drug effects , A549 Cells/microbiology , Antibodies, Bacterial/drug effects , Bordetella pertussis/pathogenicity , Cell Culture Techniques , Cell Line/drug effects , Cell Line/microbiology , Fluorescent Antibody Technique/methods , Humans , Models, Biological , Pertussis Vaccine/therapeutic use , Reproducibility of Results , Sensitivity and Specificity , Vaccination , Vaccines, Acellular/analysis , Vaccines, Acellular/therapeutic use , Whooping Cough/drug therapy , Whooping Cough/microbiology
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