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Infection and Immunity, May 1999, p. 2250-2257, Vol. 67, No. 5
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Differential Expression of Vibrio vulnificus Capsular Polysaccharide

Anita C. Wright,1,* Jan L. Powell,2 Mike K. Tanner,3 Lynne A. Ensor,2 Arthur B. Karpas,4 J. Glenn Morris Jr.,2 and Marcelo B. Sztein3

Center of Marine Biotechnology, University of Maryland Biotechnology Institute,1 and Veterans Affairs Medical Center and Department of Medicine2 and Center for Vaccine Development,3 University of Maryland School of Medicine, Baltimore, and National Institute of Child Health and Human Development, National Institutes of Health, Bethesda,4 Maryland

Received 20 August 1998/Returned for modification 9 October 1998/Accepted 23 February 1999

Vibrio vulnificus is a human pathogen whose virulence has been associated with the expression of capsular polysaccharide (CPS). Multiple CPS types have been described; however, virulence does not appear to correlate with a particular CPS composition. Reversible-phase variation for opaque and translucent colony morphologies is characterized by changes in CPS expression, as suggested by electron microscopy of cells stained nonspecifically with ruthenium red. Isolates with opaque colony morphologies are virulent and appear to be more thickly encapsulated than naturally occurring translucent-phase variants, which have reduced, patchy, or absent CPS. Previously, we have shown that the virulence of translucent-phase variants was intermediate between opaque-phase variants and acapsular transposon mutants, suggesting a correlation between virulence and the amount of CPS expressed. In the present study, CPS expression of phase variants and genetically defined mutants of V. vulnificus M06-24/O was examined by using a CPS-specific monoclonal antibody with an enzyme-linked immunosorbent assay, flow cytometry, and immunoelectron microscopy. Semiquantitative analyses of CPS expression correlated well among these assays, confirming that the translucent-phase variant was intermediate in CPS expression and retained type I CPS-specific epitopes. Cell surface expression of CPS varied with the growth phase, increasing during logarithmic growth and declining in stationary culture. Significantly greater CPS expression (P = 0.026) was observed for cells grown at 30°C than for those at 37°C. These studies confirm that phase variation and virulence in V. vulnificus correlate with the amount of CPS expressed and demonstrate the fluidity of bacterial polysaccharide expression in response to environmental conditions.

* Corresponding author. Mailing address: Center of Marine Biotechnology, University of Maryland Biotechnology Institute, 701 E. Pratt St., Baltimore, MD 21202. Phone: (410) 234-8827. Fax: (410) 234-8896. E-mail: wright{at}umbi.umd.edu.

Infection and Immunity, May 1999, p. 2250-2257, Vol. 67, No. 5
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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