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Infection and Immunity, November 2003, p. 6256-6263, Vol. 71, No. 11
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.11.6256-6263.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Identification of a Gene That Affects the Efficiency of Host Cell Infection by Legionella pneumophila in a Temperature-Dependent Fashion

Dennis A. Ridenour, Suat L. G. Cirillo, Sheng Feng, Mustapha M. Samrakandi, and Jeffrey D. Cirillo*

Department of Veterinary and Biomedical Sciences, University of Nebraska, Lincoln, Nebraska 68583

Received 2 May 2003/ Returned for modification 14 July 2003/ Accepted 23 July 2003

The ability to infect host cells is critical for the survival and replication of intracellular pathogens in humans. We previously found that many genes involved in the ability of Legionella pneumophila to infect macrophages are not expressed efficiently under standard laboratory growth conditions. We have developed an approach using expression of L. pneumophila genes from an exogenous constitutive promoter on a low-copy-number vector that allows identification of genes involved in host cell infection. Through the use of this strategy, we found that expression of a gene, lvhB2, enhances the efficiency of L. pneumophila infection of mammalian cells. The putative protein encoded by lvhB2 has similarity to structural pilin subunits of type IV secretion systems. We confirmed that this gene plays a role in host cell infection by the construction of an in-frame deletion in the L. pneumophila lvhB2 gene and complementation of this mutant with the wild-type gene. The lvhB2 mutant does not display a very obvious defect in interactions with host cells when the bacteria are grown at 37°C, but it has an approximately 100-fold effect on entry and intracellular replication when grown at 30°C. These data suggest that lvhB2 plays an important role in the efficiency of host cell infection by L. pneumophila grown at lower temperatures.

* Corresponding author. Mailing address: Dept. Veterinary and Biomedical Sciences, University of Nebraska, Lincoln, 203 VBS, Fair and East Campus Loop, Lincoln, NE 68583. Phone: (402) 472-8587. Fax: (402) 472-9690. E-mail: jcirillo1{at}unl.edu.

Editor: J. D. Clements

Infection and Immunity, November 2003, p. 6256-6263, Vol. 71, No. 11
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.11.6256-6263.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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