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Infection and Immunity, February 2007, p. 766-773, Vol. 75, No. 2
0019-9567/07/$08.00+0     doi:10.1128/IAI.00741-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Characterization of the Outer Membrane Proteome of Leptospira interrogans Expressed during Acute Lethal Infection{triangledown}

Jarlath E. Nally,1* Julian P. Whitelegge,2,3 Sara Bassilian,2,3 David R. Blanco,1 and Michael A. Lovett1

Division of Infectious Diseases, Department of Medicine,1 The Pasarow Mass Spectrometry Laboratory,2 The Jane and Terry Semel Institute for Neuroscience and Human Behavior, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, California3

Received 9 May 2006/ Returned for modification 1 June 2006/ Accepted 30 October 2006

Pathogenic Leptospira species adapt to a wide range of environmental conditions during disease transmission and infection. While the proteome of in vitro cultivated Leptospira has been characterized in several studies to date, relatively little is known of the proteome as expressed by Leptospira during disease processes. Isolates of Leptospira obtained from patients suffering the severe pulmonary form of leptospirosis cause acute lethal infection in guinea pigs and chronic asymptomatic infection in rats. Recent studies have demonstrated that protein and lipopolysaccharide constituents of Leptospira recovered from acutely infected guinea pig tissue differ from that of Leptospira in chronically infected rat tissue and in vitro cultivated Leptospira (J. E. Nally, E. Chow, M. C. Fishbein, D. R. Blanco, and M. A. Lovett, Infect. Immun. 73:3251-3260, 2005). In the current study, the proteome of Leptospira expressed during disease processes was characterized relative to that of in vitro cultivated Leptospira (IVCL) after enrichment for hydrophobic membrane proteins with Triton X-114. Protein samples were separated by two-dimensional gel electrophoresis, and antigens expressed during infection were identified by immunoblotting with monospecific antiserum and convalescent rat serum in addition to mass spectrometry. Results suggest a significant increase in the expression of the outer membrane protein Loa22 during acute infection of guinea pigs relative to other outer membrane proteins, whose expression is generally diminished relative to expression in IVCL. Significant amounts of LipL32 are also expressed by Leptospira during acute infection of guinea pigs.

* Corresponding author. Present address: School of Agriculture, Food Science and Veterinary Medicine, Veterinary Sciences Centre, University College Dublin, Belfield, Dublin 4, Ireland. Phone: 353 1 716 6182. Fax: 353 1 716 6185. E-mail: Jarlath.Nally{at}ucd.ie.

{triangledown} Published ahead of print on 13 November 2006.

Editor: W. A. Petri, Jr.

Infection and Immunity, February 2007, p. 766-773, Vol. 75, No. 2
0019-9567/07/$08.00+0     doi:10.1128/IAI.00741-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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