Pertactin evaluation as immunogen in murine model
Abstract number: 1733_1264
Rosales E., Fernandez-Ramirez A., Barcenas-Morales G., Montaraz-Crespo J.
Pertactin (PRN) is an antigen involved in protective immunity against whooping cough, caused by Bordetella pertussis. The presence of antibodies against pertactin correlates with protection in humans. B. pertussis is closely related with B. bronchiseptica, aetiological agent of respiratory tract infections in animal species. The aim of the present work was to obtain and purify pertactin from B. bronchiseptica and evaluate it as an immunogen in a murine model.
Previously, infection murine model has been employed using three different mouse strains (NIH, Hsd:ICR and BALB/c) and four B. bronchiseptica strains by aerosol via. In the present work, three methods were tested in order to obtain PRN from two different B. bronchiseptica strains: I. By sonicate and acetone precipitation; II. By acid extraction method (pH = 3) and, III. By ultracentrifugal series and ethanol precipitation. The product of each method was corroborated by SDS-PAGE and western blot (BB07 and BB05 monoclonal antibodies) assays. Groups of mice were immunised with PRN, PRN+FCA or B. bronchiseptica bacterin (day zero) and immunisations were repeated at day 15. All mice were challenged with B. bronchiseptica at 21st day; cfu/lung were determined at days 0, 3, 5, 7, 10, 15 and 21 after challenge. Additionally, the two groups of immunised mice with each pertactin preparation was tested in a cross-challenge experiment.
Hsd:ICR and BALB/c mouse strains were susceptible to infection by LBF (isolated from pig) and ESP1 (from human) B. bronchiseptica strains, respectively. PRN was found to be protective for both mouse strains, and this protection was even better than that conferred by whole cell bacterin. Finally, cross protection effect was not observed when the animals were immunised with the pertactin preparations.
In conclusion, the animal test described in this work could be considered as a good option for future studies of comparison the protection grade using different commercial pertactin vaccines.
|Session name:||European Society of Clinical Microbiology and Infectious Diseases|
|Location:||ICC, Munich, Germany|
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