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Infection and Immunity, August 1999, p. 4243-4250, Vol. 67, No. 8
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Protection against Mycobacterium avium by DNA Vaccines Expressing Mycobacterial Antigens as Fusion Proteins with Green Fluorescent Protein

Maria Velaz-Faircloth,1,2 Alison J. Cobb,2 Amanda L. Horstman,3 Stanley C. Henry,1 and Richard Frothingham1,2,4,*

Infectious Disease Section, Veterans Affairs Medical Center, Durham, North Carolina 27705,1 and Departments of Medicine,2 Biochemistry,3 and Microbiology,4 Duke University Medical Center, Durham, North Carolina 27710

Received 14 December 1998/Returned for modification 21 January 1999/Accepted 3 May 1999

Mycobacterium avium causes disseminated disease in humans with AIDS, paratuberculosis in ruminants, lymphadenopathy in swine, and tuberculosis in birds. We constructed DNA vaccines expressing mycobacterial antigens as fusion proteins with enhanced green fluorescent protein (EGFP). Plasmids p65K-EGFP, p85A-EGFP, and p85B-EGFP expressed the M. avium 65-kDa antigen, the Mycobacterium bovis BCG 85A antigen, and the M. avium 85B antigen, respectively, as EGFP fusion proteins. We visualized protein expression directly in cultured murine fibroblasts and intact muscle. p65K-EGFP expressed fusion protein in a diffuse cytoplasmic pattern, and p85A-EGFP and p85B-EGFP produced a speckled pattern. We vaccinated C57BL/6 mice with three doses of plasmid DNA and then challenged them intraperitoneally with M. avium. Negative controls received saline, and positive controls received one dose of BCG vaccine. Mice in all groups developed disseminated infection with a high burden of organisms. Compared to negative controls, mice vaccinated with p85A-EGFP had an eightfold reduction in spleen M. avium CFU at 4 weeks after infection and a fourfold reduction at 8 weeks, reductions similar to those generated by BCG vaccine. Mice vaccinated with p65K-EGFP had a fourfold CFU reduction at 4 weeks and no effect at 8 weeks. This is the first report of DNA vaccines expressing foreign antigens as fusion proteins with EGFP and the first report of successful DNA vaccination against M. avium.

* Corresponding author. Mailing address: Veterans Affairs Medical Center, 508 Fulton St., Building 4, Durham, NC 27705. Phone: (919) 286-0411, ext. 6566. Fax: (919) 286-6824. E-mail: richard.frothingham{at}duke.edu.

Infection and Immunity, August 1999, p. 4243-4250, Vol. 67, No. 8
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

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