1. Pasteur Institute of Iran 2. Baqiyatallah University of Medical Sciences 3. Pasteur Institute of Iran
Abstract
Introduction
Brucellosis is the most common zoonotic bacterial disease that is transmitted from animals to humans. a key tool for the control, elimination, and eradication of brucellosis is the development of an effective vaccine; however, there is no available effective vaccine against brucella. in this work, we evaluated the vaccine potential of modified chitosan nanoparticles containing flic protein against brucellosis.
Methods
The amplification of flic gene was performed by using the pcr. then, the amplified gene was cloned and expressed in pet28a-bl21 (de3) expression system. the proteins were purified with ni-nta column. sds-page and western blot was used for confirmation of purified protein. mannosylated chitosan nanoparticles are synthesized by chemical synthesis and then nanoparticles are examined in terms of structure, shape and size by ftir, sem and dls. mannosylated chitosan nanoparticles containing flic were applied for immunization of balb/c mice. antibody-dependent immune mechanisms as well as the ifn-γ, il-2, and il-10 cytokines were determined in post-immunized mouse serum. in challenging tests different groups of immunized mice were infected with brucella.
Results
Flic gene was amplified and cloned in pet28a vector. then, expression of flic was performed in bl21 (de3) that sds-page and western blot showed the high purity of the eluted protein on column. s.c immunization of mannosylated chitosan nanoparticles containing flic elicited a strong specific igg response (higher igg2a titers) and significant ifn-γ /il2 production.
Conclusion
In general, the results exhibited that flic is immunogenic antigen and its administration with mannosylated chitosan in mice induced th1 immune response.
