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Infection and Immunity, September 2002, p. 5193-5201, Vol. 70, No. 9
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.9.5193-5201.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Analysis of Pathogen-Host Cell Interactions in Purpura Fulminans: Expression of Capsule, Type IV Pili, and PorA by Neisseria meningitidis In Vivo

O. B. Harrison,1 B. D. Robertson,1 S. N. Faust,2 M. A. Jepson,3 R. D. Goldin,4 M. Levin,2 and R. S. Heyderman5*

Infectious Diseases & Microbiology,1 Paediatrics,2 Histopathology, Faculty of Medicine, Imperial College, London,4 Pathology & Microbiology,5 Biochemistry, University of Bristol, Bristol, United Kingdom3

Received 21 March 2002/ Returned for modification 25 April 2002/ Accepted 17 June 2002

The pattern of meningococcal surface structure expression in different microenvironments following bloodstream invasion in vivo is not known. We used immunohistochemistry to determine the expression of capsule, type IV pili, and PorA by meningococci residing in the skin lesions of children with purpura fulminans. All the skin biopsy samples showed evidence of thrombosis and, frequently, a perivascular inflammatory cell infiltrate consisting of neutrophils (elastase positive) and monocytes/macrophages (CD68 positive). Modified Gram staining revealed 20 to over 100 gram-negative diplococci in each 4-µm-thick section, usually grouped into microcolonies. Immunoperoxidase staining demonstrated that the invading meningococci expressed PorA, capsule, and type IV pilin. Expression of these antigens was not restricted to any particular environment and was found in association with meningococci located in leukocytes, small blood vessels, and the dermal interstitium. Confocal laser scanning microscopy demonstrated coexpression of pilin and capsule by numerous microcolonies. However, there was some discordance in capsule and pilin expression within the microcolonies, suggesting phase variation. The strategy employed in this study will be helpful in investigating invasive bacterial diseases where antigenic and phase variation has a significant impact on virulence and on vaccine design.

* Corresponding author. Mailing address: Department of Pathology & Microbiology, School of Medical Sciences, University of Bristol, University Walk, Bristol BS8 1TD, United Kingdom. Phone: 44 (0) 117 928 2567. Fax: 44 (0) 117 929 9162. E-mail: r.heyderman{at}bristol.ac.uk.

Editor: D. L. Burns

Infection and Immunity, September 2002, p. 5193-5201, Vol. 70, No. 9
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.9.5193-5201.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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