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Infection and Immunity, March 2004, p. 1423-1430, Vol. 72, No. 3
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.3.1423-1430.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

The Klebsiella pneumoniae O Antigen Contributes to Bacteremia and Lethality during Murine Pneumonia

Sunita Shankar-Sinha,1,{dagger} Gabriel A. Valencia,1,{dagger} Brian K. Janes,2 Jessica K. Rosenberg,1 Chris Whitfield,3 Robert A. Bender,2 Ted J. Standiford,4 and John G. Younger1*

Departments of Emergency Medicine,1 Internal Medicine, School of Medicine,4 Department of Molecular, Cellular, and Developmental Biology, School of Literature, Science, and Arts, University of Michigan, Ann Arbor, Michigan,2 Department of Microbiology, University of Guelph, Guelph, Ontario, Canada3

Received 8 May 2003/ Returned for modification 16 July 2003/ Accepted 21 November 2003

Bacterial surface carbohydrates are important pathogenic factors in gram-negative pneumonia infections. Among these factors, O antigen has been reported to protect pathogens against complement-mediated killing. To examine further the role of O antigen, we insertionally inactivated the gene encoding a galactosyltransferase necessary for serotype O1 O-antigen synthesis (wbbO) from Klebsiella pneumoniae 43816. Analysis of the mutant lipopolysaccharide by sodium dodecyl sulfate-polyacrylamide gel electrophoresis confirmed the absence of O antigen. In vitro, there were no detectable differences between wild-type K. pneumoniae and the O-antigen-deficient mutant in regard to avid binding by murine complement C3 or resistance to serum- or whole-blood-mediated killing. Nevertheless, the 72-h 50% lethal dose of the wild-type strain was 30-fold greater than that of the mutant (2 x 103 versus 6 x 104 CFU) after intratracheal injection in ICR strain mice. Despite being less lethal, the mutant organism exhibited comparable intrapulmonary proliferation at 24 h compared to the level of the wild type. Whole-lung chemokine expression (CCL3 and CXCL2) and bronchoalveolar inflammatory cell content were also similar between the two infections. However, whereas the wild-type organism produced bacteremia within 24 h of infection in every instance, bacteremia was not seen in mutant-infected mice. These results suggest that during murine pneumonia caused by K. pneumoniae, O antigen contributes to lethality by increasing the propensity for bacteremia and not by significantly changing the early course of intrapulmonary infection.

* Corresponding author. Mailing address: 7679 Kresge Research Building I, 200 Zina Pitcher Pl., Ann Arbor, MI 48109-0303. Phone: (734) 647-7564. Fax: (734) 615-4220. E-mail: jyounger{at}umich.edu.

Editor: J. N. Weiser

{dagger} S.S.-S. and G.A.V. contributed equally as first authors to this work.

Infection and Immunity, March 2004, p. 1423-1430, Vol. 72, No. 3
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.3.1423-1430.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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