Immunization of Pseudomonas aeruginosa alginate - conjugated to diphteria toxoid in mice

Subject Areas : Molecular Microbiology

Ghazaleh Jaberi ¹ , Reza Shapouri ² , Ashraf Kariminik ³

1 - M.Sc., Department of Microbiology, Science and Research Branch, Islamic Azad University, Kerman, Iran
2 - Assistant Professor, Department of Microbiology, Zanjan Branch, Islamic Azad University, Zanjan, Iran
3 - Lecturer, Department of Microbiology, Kerman Branch, Islamic Azad University, Kerman, Iran.

Received: 2013-04-01 Accepted : 2013-05-05 Published : 2013-06-01

Keywords: ELISA, Alginate, Diphtheria toxoid, Conjugate vaccine, Pseudomonas aeroginosa,

Abstract :

Background and Objectives: Pseudomonas aeruginosa is one of the most important infectious agent in medicine. These bacteria cause several infections especially in the patient with high degrees of burns and in immunodeficiency cases. The purpose of this study was providing conjugated antigen component that can induct production of antibody and memory immune in mouse model against Pseudomonas aeruginosa.  Materials and Methods: After culturing P. aeruginosa strain PAO1 and extraction of alginate, Diphteria toxoid, ADH and EDAC were added as protein carrier, spacer and linker, respectively to the alginate. After passing the Alginate diphtheria toxoid conjugated (ALG-DT) through chromatography column (CL-2B), its quality was checked to get the quality control label. The prepared antigens were intraperitoneally injected to BALB/c mice (10µg/ml/mouse) for three times once each two weeks. Blood sampling was performed after two weeks of each injection and the levels of serum IgG, IgA and IgM were measured by ELISA. Results: The titer of serum antibodies of vaccinated group with ALG-DT increased significantly after each injection. The levels of serum IgG, IgM and IgA against alginate in vaccinated group was significantly more than the control groups, and after third injection reached to 3.5, 1.7 and 1.2 times increases, respectively. Conclusion: The increases in the levels of  antibodies in vaccinated groups are an indicator of activation of T-cells and memory cells. As a result, the conjugated alginate-diphtheria toxoid can be appropriate candidates for production of vaccine.