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Chapter 64 : Lipopolysaccharide Architecture of Grown in Broth and Host Cells

Authors: Jürgen H. Helbig1, Esteban Fernandez-Moreira2, Enno Jacobs3, Paul Christian Lück4, Martin Witt5
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Affiliations: 1: Medical Faculty TU Dresden, Institute of Medical Microbiology and Hygiene, D-01307 Dresden, Germany; 2: University of Michigan Medical School, Depart ment of Microbiology and Immunology, Ann Arbor, MI, 48109; 3: Medical Faculty TU Dresden, Institute of Medical Microbiology and Hygiene, D-01307 Dresden, Germany; 4: Medical Faculty TU Dresden, Institute of Medical Microbiology and Hygiene, D-01307 Dresden, Germany; 5: Medical Faculty TU Dresden, Institute of Anatomy, D-01307 Dresden, Germany
Content Type: Monograph
Publication Year: 2006

Category: Bacterial Pathogenesis

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Abstract:

This chapter demonstrates for the first time to what extent the lipopolysaccharide (LPS) architecture is involved in extracellular structures of bacteria. Furthermore, the intraphogosomal/ intracellular shedding of LPS components is investigated. The authors' investigation of the surface architecture of strain Lp02 and its and mutants substantiate that the LPS equipment depends at least on both genes. Therefore, the valuation of the influence of and on pathogenicity also has to consider the phenotypic changes of the LPS surface structures, because they initiate the first step of interaction between bacteria and host cells. The amounts of LPS were quantified by enzyme-linked immunosorbent assay. LPS was immobilized in microtiter wells and detected by monoclonal antibody (MAb) 3/1 followed by antimouse-IgG horseradish peroxidase. Moreover, until the postexponential growth phase in broth cultures only approximately 5% of bacteria of strains Corby and Lp02 can be labeled by MAb 59/1. After lysis of host cells it was observed that legionellae were released either as bacterial clusters positive for both MAbs or as single MAb 3/1-positive bacteria being MAb 59/1-positive or negative. The ability to modify and shed LPS allows to build lamellar structures and vesicles and to release soluble LPS that may modulate host cells, enhance survival in aerosols, or stabilize biofilms. In this chapter, the authors have demonstrated that the whole LPS architecture is influenced by at least two systems, namely the type IV secretion apparatus and regulatory elements.

Citation: H. Helbig J, Fernandez-Moreira E, Jacobs E, Christian Lück P, Witt M. 2006. Lipopolysaccharide Architecture of Grown in Broth and Host Cells, p 261-264. In Cianciotto N, Kwaik Y, Edelstein P, Fields B, Geary D, Harrison T, Joseph C, Ratcliff R, Stout J, Swanson M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815660.ch64
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Lipopolysaccharide Architecture of Legionella pneumophila Grown in Broth and Host Cells
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Citation: H. Helbig J, Fernandez-Moreira E, Jacobs E, Christian Lück P, Witt M. 2006. Lipopolysaccharide Architecture of Grown in Broth and Host Cells, p 261-264. In Cianciotto N, Kwaik Y, Edelstein P, Fields B, Geary D, Harrison T, Joseph C, Ratcliff R, Stout J, Swanson M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815660.ch64
/content/10.1128/9781555815660.ch64.ch64fig01
FIGURE 1
Shedding of LPS in broth ( to ) and in cells 6 h () or 19 h () after infection with strain Corby. LPS structures were labeled with MAb 3/1 ( to ) or double-labeled with MAb 59/1 plus MAb 3/1 ( and ). Objects positive for both antibodies appear white; only MAb 3/1-positive structures appear grey. MAb 59/1-positive only was not obtained. Exp, exponential growth phase; PE, postexponential growth phase. For details, see text.
10.1128/9781555815660/f0262-01_thmb.gif
10.1128/9781555815660/f0262-01.gif
Image of FIGURE 1
FIGURE 1

Shedding of LPS in broth ( to ) and in cells 6 h () or 19 h () after infection with strain Corby. LPS structures were labeled with MAb 3/1 ( to ) or double-labeled with MAb 59/1 plus MAb 3/1 ( and ). Objects positive for both antibodies appear white; only MAb 3/1-positive structures appear grey. MAb 59/1-positive only was not obtained. Exp, exponential growth phase; PE, postexponential growth phase. For details, see text.

Citation: H. Helbig J, Fernandez-Moreira E, Jacobs E, Christian Lück P, Witt M. 2006. Lipopolysaccharide Architecture of Grown in Broth and Host Cells, p 261-264. In Cianciotto N, Kwaik Y, Edelstein P, Fields B, Geary D, Harrison T, Joseph C, Ratcliff R, Stout J, Swanson M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815660.ch64
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/content/10.1128/9781555815660.ch64.ch64fig02
FIGURE 2
Transmission electron micrograph (A) and immunogold labeling (B, C) of U937 cells infected by strain Corby (14 h postinfection). The LPS was labeled with MAb 3/1. Distinguishing, large white arrows; phagosome membrane ( and ), small black arrows; LPS on bacterial surface (), ellipse; shed LPS crossing the phagosomal membrane (), circle; shed LPS inside the cytoplasm ().
10.1128/9781555815660/f0263-01_thmb.gif
10.1128/9781555815660/f0263-01.gif
Image of FIGURE 2
FIGURE 2

Transmission electron micrograph (A) and immunogold labeling (B, C) of U937 cells infected by strain Corby (14 h postinfection). The LPS was labeled with MAb 3/1. Distinguishing, large white arrows; phagosome membrane ( and ), small black arrows; LPS on bacterial surface (), ellipse; shed LPS crossing the phagosomal membrane (), circle; shed LPS inside the cytoplasm ().

Citation: H. Helbig J, Fernandez-Moreira E, Jacobs E, Christian Lück P, Witt M. 2006. Lipopolysaccharide Architecture of Grown in Broth and Host Cells, p 261-264. In Cianciotto N, Kwaik Y, Edelstein P, Fields B, Geary D, Harrison T, Joseph C, Ratcliff R, Stout J, Swanson M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815660.ch64
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References

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1. Helbig, J. H.,, P. C. Lück,, Y. A. Knirel,, W. Witz-leb, and, U. Zähringer. 1995. Molecular characterization of a virulence-associated epitope on the lipopolysaccharide of Legionella pneumophila serogroup 1. Epidemiol. Infect. 115: 71— 78.
2. Helbig, J. H.,, S. Bernander,, M. Castellani Pastoris,, J. Etienne,, V. Gaia,, S. Lauwers,, D. Lindsay,, P. C. Lück,, T. Marques,, S. Mentula,, M. F. Peeters,, C. Pelaz,, M. Struelens,, S. A. Uldum,, G. Wewalka, and, T. G. Harrison. 2002. Pan-European study on culture-proven Legionnaires’ disease: distribution of Legionella pneumophila serogroups and monoclonal subgroups. Eur. J. Clin. Microbiol Infect. Dis. 21: 710— 716.
3. Lüneberg, E.,, B. Meyer,, N. Daryab,, O. Koois-tra,, U. Zähringer,, M. Rhode,, J. Swanson, and, M. Frosch. 2001. Chromosomal insertion and excision of a 30 kb unstable genetic element is responsible for phase variation of lipopolysaccha-ride and other virulence determinants in Legionella pneumophila. Mol. Microbiol. 39: 1259— 1271.
4. Lüneberg, E.,, U. Zähringer,, Y. A. Knirel,, D. Steinmann,, M. Hartmann,, I. Steinmetz,, M. Rohde,, J. Köhl, and, M. Frosch. 1998. Phasevariable expression of lipopolysaccharide contributes to virulence of Legionella pneumophila. J. Exp. Med. 188: 49— 60.
5. Swanson, M. S., and, E. Fernandez-Moreia. 2002. A microbial strategy to multiply in macrophages: the pregnant pause. Traffic 3: 170— 77.
6. Zou, C. H.,, Y. A. Knirel,, J. H. Helbig,, U. Zähringer, and, C. S. Mintz. 1999. Molecular cloning and characterization of a locus responsible for O-acetylation of the O polysaccharide of Legionella pneumophila serogroup 1 lipopolysac-charide. J. Bacteriol. 181: 4137— 4141.

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