Outer membrane vesicles as acellular vaccine against pertussis.

ROBERTS ROY M; MORENO GRISELDA; BOTTERO DANIELA; GAILLARD M. EMILIA; FINGERMANN MATÍAS; GRIAEB AUGUSTO; RUMBO MARTÍN; HOZBOR DANIELA

VACCINE

Elsevier

Año: 2008 p. 4639 - 4646

0264-410X

In this study the development and evaluation of outer membrane vesicles (OMVs) obtained from Bordetella pertussis as vaccines against pertussis disease is described. SDS-PAGE, immunoblot techniques and gel electrophoresis associated to tandem mass spectrometry were used to describe the composition of the OMVs obtained in our laboratory from B. pertussis Tohama CIP 8132 strain. These techniques revealed the presence of the main well known pertussis surface immunogens in the OMVs such as pertactin, adenylate cyclase ? hemolysin, pertussis toxin, as well as the lipopolysaccharide (LPS). Furthermore, by matrix-assisted laser desorption ionization?time-of-flight mass spectrometry analysis a total of 43 proteins were identified. Some of the proteins identified were predicted to have outer membrane or periplasmic location and relatively few cytoplasmic membrane proteins were detected. The proteins not predicted to have an outer membrane location were predominantly cytoplasmic or had an unknown location. The characterized pertussis OMVs were used in murine B. pertussis intranasal challenge model to examine their protective capacity. Killed detoxified whole cell B. pertussis bacteria were used as control. Significant differences between intraperitoneal immunized BALB/c and the control group vaccinated with PBS were observed (p<0.001). Adequate elimination rates (p<0.005) were observed in mice immunized either with OMV or whole cell bacteria. Mice were also protected against pertussis when they were intranasally immunized with OMVs. Comparable protection results were obtained with both type of immunization route. Thus intranasal treatment with OMVs enhanced markers of innate immune response such as TNFa, IL-6 and CCL20 at early time points (2 to 8 hours) as well as whole cell bacteria. In view to their capacity to induce airways innate and protective immunity in the mouse model, OMVs obtained from B pertussis are candidates to be used not only as intraperitoneal vaccine but also as intranasal vaccine to protect against pertussis.