ISOLATION AND CHARACTERIZATION OF OUTER MEMBRANE VESICLES (OMVS) FROM BORDETELLA PERTUSSIS TOHAMA STRAIN

Background and Aim:Despite huge vaccination against pertussis, it is a global health problem because of low efficacy of aP (acellular vaccine) that substituted with reactogenic wP (whole cell vaccine). So a third generation of pertussis vaccine with good safety and potency is needed. Outer membrane vesicles (OMVs) of Bordetella pertussis contain several immunogens and they are good candidate for vaccine production. The main target of this study is to extract and characterization of OMVs from B. pertussisMethods:Firstly the OMVs isolated by sequential ultracentrifugation. After negative phosphotungstate potassium staining the shape and size of extracted OMVs analysed by electron microscopy studies. Protein content measured by Bradford method and protein profiles studied by SDS-PAGE assay.Results:The Size ranging of obtained OMVs was between 40 to 200nm. The protein content of the OMVs was 150 µg/ml and the protein profiles of extracted OMVs studied by SDS-PAGE indicated that there is several proteins in the OMVs.Conclusion:Sveral resurgences of pertussis reported in several countries that shifted to aP from wP. Because of the side-effects associated with wP and low efficacy of aP, it is crucial to design a potent and safe vaccine against pertussis. OMVs of B.pertussis contains main immunogens and can induce Th1, Th2, Th17 as similar as wP. So the OMVs are good vaccine candidate for pertussis. The results showed that obtained OMVs were spherical nanoparticles and contain several proteins and can be used as acellular vaccine after further inestigation.

کلیدواژه ها:

Bordetella pertussis ، Outer membrane vesicle ، acellular vaccine ، whole cell vaccine