Fatemeh Vahedi; Mahmoud Reza Jaafari; Mahmoud Mahmoudi
Volume 7, Issue 4 , December 2010, , Pages 210-216
Abstract
Background: DNA vaccines are third generation vaccines which have made promises to combat infectious diseases. Cationic liposomes are used as effective delivery systems for DNA vaccines to generate stronger immunity. Objective: Encapsulation of pcDNA3.1+PA plasmid, encoding protective antigen (PA) of ...
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Background: DNA vaccines are third generation vaccines which have made promises to combat infectious diseases. Cationic liposomes are used as effective delivery systems for DNA vaccines to generate stronger immunity. Objective: Encapsulation of pcDNA3.1+PA plasmid, encoding protective antigen (PA) of Bacillus anthracis (B. anthracis) into cationic liposomes, and evaluation of its effect on specific humoral specific immunity against PA were aimed. Methods: The liposomes containing pcDNA3.1+PA plasmids were prepared with phosphatidylcholine (PC), dioleoyl phosphatidylethanolamine (DOPE) and 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) using dehydration-rehydration method. BALB/c mice were immunized by intramuscular (IM) injection to investigate the immunogenicity of the formulations. The resulting specific antibodies against PA, total IgG, IgG1, IgG2a and IgG2b isotypes, were evaluated by enzyme linked immunosorbent assay (ELISA) method. Conclusion: A higher concentration of specific IgG against PA was found in sera of a group immunized with the encapsulated plasmid compared with the naked plasmid alone. This difference was significant for IgG1 isotype.
Fatemeh Vahedi; Naser Taiebi Meibody; Mahdi KianiZadeh; Mahmoud Mahmoudi
Volume 2, Issue 3 , September 2005, , Pages 134-140
Abstract
Background: DNA immunization with plasmid DNA encoding bacterial, viral, parasitic and tumor antigens has been reported to trigger protective immunity. Objective: To evaluate the use of a DNA immunization strategy for protection against anthrax, a plasmid was constructed. Methods: The partialsequence ...
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Background: DNA immunization with plasmid DNA encoding bacterial, viral, parasitic and tumor antigens has been reported to trigger protective immunity. Objective: To evaluate the use of a DNA immunization strategy for protection against anthrax, a plasmid was constructed. Methods: The partialsequence of protective antigen of Bacillus anthracis, amino acids 175-764, as a potent immunogenic target was selected. The DNA encoding this segment was utilized in the construction of pcDNA3.1+PA plasmid. After intramuscular injection of rats with pcDNA3.1+PA plasmid, the expression of PA was assessed by RT-PCR and immunohistochemistry at RNA and protein levels, respectively. We also evaluated the presence of anti-PA antibodies in sera of immunized mice with pcDNA3.1+PA construct using immunoblotting. Results: The integrity of pcDNA3.1+PA construct was confirmed with restriction analysis and sequencing. The expression of PA was detected at RNA and protein levels. The presence of anti-PA antibodies in immunized mice with pcDNA3.1+PA construct was also confirmed. Conclusion: Our results indicate that pcDNA3.1+PA eukaryotic expressing vector could express PA antigen, induce antibody response and may be used as a candidate for DNA vaccine against anthrax.