title: A novel high-throughput assay to quantify the vaccine-induced inhibition of Bordetella pertussis adhesion to airway epithelia.
abstract: 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.
  
  
  
  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.
  
  
  
  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.
authors.affiliation: GSK Vaccines, Via Fiorentina 1, 53100, Siena, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Zanaboni
authors.affiliation: GSK Vaccines, Via Fiorentina 1, 53100, Siena, Italy.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Arato
authors.affiliation: GSK Vaccines, Via Fiorentina 1, 53100, Siena, Italy.
authors.forename: Mariagrazia
authors.initials: M
authors.lastname: Pizza
authors.affiliation: GSK Vaccines, Via Fiorentina 1, 53100, Siena, Italy.
authors.forename: Anja
authors.initials: A
authors.lastname: Seubert
authors.affiliation: GSK Vaccines, Via Fiorentina 1, 53100, Siena, Italy. rosanna.x.leuzzi@gsk.com.
authors.forename: Rosanna
authors.initials: R
authors.lastname: Leuzzi
mesh_terms: D000072283:A549 Cells / Q000187:drug effects / Q000382:microbiology; D018829:Adhesins, Bacterial / Q000032:analysis*; D000907:Antibodies, Bacterial / Q000187:drug effects; D001886:Bordetella pertussis / Q000187:drug effects* / Q000472:pathogenicity; D018929:Cell Culture Techniques; D002460:Cell Line / Q000187:drug effects / Q000382:microbiology; D004847:Epithelial Cells / Q000382:microbiology*; D005455:Fluorescent Antibody Technique / Q000379:methods; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D008954:Models, Biological; D010567:Pertussis Vaccine / Q000032:analysis* / Q000627:therapeutic use; D015203:Reproducibility of Results; D012137:Respiratory System / Q000382:microbiology*; D012680:Sensitivity and Specificity; D014611:Vaccination; D022282:Vaccines, Acellular / Q000032:analysis / Q000627:therapeutic use; D014917:Whooping Cough / Q000188:drug therapy / Q000382:microbiology
chemical_list: D018829:Adhesins, Bacterial; D000907:Antibodies, Bacterial; D010567:Pertussis Vaccine; D022282:Vaccines, Acellular
keywords: Adhesion assay; Airway epithelium; B. pertussis; Fluorescent-labeled bacteria