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Research Article

ScienceAsia 30 (2004): 231-237 |doi: 10.2306/scienceasia1513-1874.2004.30.231

Characterization and Protection in Mice of Outer Membrane Proteins from Pasteurella multocida A:1 Incorporated in Lipid Vaccine Delivery Systems

Atthachai Homhuan,a* Pornpen Pathanasophon,b Daan JA Crommelin,c Wim Jiskoot,c Gideon FA Kerstend and Sompol Prakongpana

ABSTRACT: Fowl cholera is an infectious disease affecting poultry and is caused by Pasteurella multocida. To develop a subunit vaccine, outer membrane proteins (OMPs) from P. multocida serotype A:1 strain NIAH DU1551/97 were extracted and characterized using SDS-PAGE. The OMPs were solubilized in detergent or incorporated in lipid-based antigen delivery systems, namely virosomes made from Newcastle disease (ND) virus and immunostimulating complexes (ISCOMs). The formulations were characterized by several physicochemical methods. To evaluate the formulations as possible vaccine, their potency was tested in mice. Eleven proteins that range in size from 30 to 80 kDa were detected in the OMP fraction. The most prominent protein bands were 30, 33, and 45 kDa. Virosomes and ISCOMs showed an average diameter of 180 and 30 nm, respectively. OMPs incorporated in the virosomal membrane inhibited the capability of virosomes to agglutinate chicken red blood cells. Animals were challenged after two immunizations. All vaccines fully protected mice against a low dose challenge. In the case of a high dose challenge, micellar OMPs provided 80% protection, whereas OMPs incorporated in virosomes or ISCOMs gave 100% protection, which is comparable to that of inactivated whole cell vaccines. In conclusion, antigen loaded virosomes and ISCOMs are potential pasteurella subunit vaccines.

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a Department of Pharmacy, Faculty of Pharmacy, Mahidol University, Bangkok 10400, Thailand.
b Bacteriology Section, National Institute of Animal Health, Bangkok 10900, Thailand.
c Department of Pharmaceutics, Faculty of Pharmaceutical Sciences, Utrecht Institute for Pharmaceutical Sciences (UIPS), Utrecht University, P.O. Box 80082, 3508 TB Utrecht, The Netherlands.
d Research and Development Unit, Netherlands Vaccine Institute, P.O. Box 457, 3750 AL Bilthoven, The Netherlands.

* Corresponding author, E-mail: g4138458@yahoo.com

Received 20 Jan 2004, Accepted 21 May 2004