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CpG-Modified Plasmid DNA Encoding Flagellin Improves Immunogenicity and Provides Protection against Burkholderia pseudomallei Infection in BALB/c Mice

Section of Infectious Disease, Kaohsiung Veterans General Hospital, Kaohsiung, Taiwan,¹ Institute of Environmental Education, National Kaohsiung Normal University, Kaohsiung, Taiwan,² Department of Biotechnology, National Kaohsiung Normal University, Kaohsiung, Taiwan,³ Department of Infectious Disease, E-DA Hospital/I-Shou University, Kaohsiung, Taiwan,⁴ Department of Microbiology and Immunology, University of Arkansas for Medical Sciences, Little Rock, Arkansas⁵

Received 13 August 2005/ Returned for modification 22 September 2005/ Accepted 15 December 2005

The plasmid DNA encoding the fliC gene of Burkholderia pseudomallei combined with CpG oligodeoxynucleotide (ODN) was injected intramuscularly into BALB/c mice, resulting in the increased production of certain humoral antibodies and flagellin-specific spleen cell clonal expansion. CpG ODN, as an immunoadjuvant, was added to the plasmid containing the fliC gene in order to obtain ongoing expression in muscle for a long period. Functional expression of flagellin from the constructed CpG-modified plasmid in transfected peritoneal exudate cells of BALB/c mice was shown by reverse transcription-PCR and Western blotting. Furthermore, BALB/c mice immunized with the modified plasmid had relatively higher resistance to B. pseudomallei infection in vivo than did mice immunized with unmodified plasmid DNA. The time course of restricted bacterial growth in spleen and liver and changes in the cytokine profiles of immunized mice suggested that the stimulated phagocytic cells would be able to kill the bacteria eventually, possibly as a consequence of the induction of Th-1-type immune polarization in vivo. Th-1-type immune polarization was detected in response to flagellin induction in mice immunized with CpG-modified plasmid DNA by the appearance of increased levels of immunoglobulin G2a antibodies and gamma interferon-secreting cells specific to flagellin. The exogenous CpG motifs added to the fliC gene would contribute to an adjuvant-like response that enhances the flagellin-specific immunogenicity and provides protection against B. pseudomallei infection. This CpG-modified plasmid DNA vaccination is an important potential strategy that should be developed to protect against melioidosis.

Editor: J. B. Bliska

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