Chemically defined medium for Legionella pneumophila growth

Ristroph, Joseph D.; Hedlund, Kenneth; Gowda, S. Veerana

doi:10.1128/jcm.13.1.115-119.1981

Cited by 84 publications

(63 citation statements)

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“…Moreover, determination of the intracellular growth rate by gentamicin kill assay demonstrated ,1000-fold increased bacterial numbers within 48 h, suggesting a generation time of at least ,5 h that was similar to other Proteobacteria, e.g. Rickettsia spp., Anaplasma marginale or Legionella pneumophila ranging from 4 to 17 h (Hidalgo, 1975;Ristroph et al, 1981;Pang and Winkler, 1994). Therefore, B. henselae can be considered as a facultative intracellular pathogen.…”

Section: Discussionmentioning

confidence: 99%

Interaction of Bartonella henselae with endothelial cells results in rapid bacterial rRNA synthesis and replication

et al. 2000

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Bartonella henselae is a slow‐growing microorganism and the causative pathogen of bacillary angiomatosis in man. Here, we analysed how interaction of B. henselae with endothelial cells might affect bacterial growth. For this purpose, bacterial rRNA production and ribosome content was determined by fluorescence in situ hybridization (FISH) using rRNA‐targeted fluorescence‐labelled oligonucleotide probes. B. henselae grown on agar plates showed no detectable rRNA content by means of FISH, whereas B. henselae co‐cultured with endothelial cells showed a rapid increase of rRNA production within the first 18 h after inoculation. The increased rRNA synthesis was paralleled by a ∼1000‐fold intracellular bacterial replication, whereas bacteria grown on agar base showed only a ∼10‐fold replication within the first 48 h of culture. Pretreatment of host cells with paraformaldehyde prevented adhesion, invasion, intracellular replication and bacterial rRNA synthesis of B. henselae. In contrast, inhibition of host cell protein synthesis by cycloheximide did not affect bacterial adhesion and invasion, but prevented intracellular replication although bacterial rRNA content was increased. Inhibition of actin polymerization by cytochalasin D did not affect adhesion, invasion, increased rRNA content or intracellular replication of B. henselae. These results demonstrate that rRNA synthesis and replication of B. henselae is promoted by viable host cells with intact de novo protein synthesis.

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Section: Discussionmentioning

confidence: 99%

Interaction of Bartonella henselae with endothelial cells results in rapid bacterial rRNA synthesis and replication

et al. 2000

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“…Legionella pneumophila relies on amino acids as the main source of energy and carbon, although carbohydrates may be a minor substrate (George et al, 1980;Ristroph et al, 1981;Tesh and Miller, 1981;Eylert et al, 2010). Even though L. pneumophila preferentially utilizes amino acids for carbon and energy, in silico analysis of the L. pneumophila genome sequence revealed that this organism is auxotrophic for the seven amino acids Arg, Cys, Ile, Leu, Met, Thr and Val (Cazalet et al, 2004;Chien et al, 2004).…”

Section: Amino Acid Requirements Of L Pneumophilamentioning

confidence: 99%

“…Even though L. pneumophila preferentially utilizes amino acids for carbon and energy, in silico analysis of the L. pneumophila genome sequence revealed that this organism is auxotrophic for the seven amino acids Arg, Cys, Ile, Leu, Met, Thr and Val (Cazalet et al, 2004;Chien et al, 2004). Furthermore, it has been shown that the amino acids Arg, Cys, Ile, Leu, Met, Ser, Thr and Val are essential for growth of L. pneumophila in vitro (Pine et al, 1979;George et al, 1980;Reeves et al, 1981;Ristroph et al, 1981;Tesh and Miller, 1981;Tesh et al, 1983;Molofsky and Swanson, 2004;Wieland et al, 2005;Eylert et al, 2010). The four amino acids, Arg, Cys, Gln and Ser, are needed to support growth in vivo (Wieland et al, 2005).…”

Section: Amino Acid Requirements Of L Pneumophilamentioning

confidence: 99%

“…Even though L. pneumophila absolutely requires amino acids to power intracellular growth, it is auxotrophic for Arg, Cys, Ile, Leu, Met, Thr and Val (Fig. 3) (Pine et al, 1979;George et al, 1980;Reeves et al, 1981;Ristroph et al, 1981;Tesh and Miller, 1981;Tesh et al, 1983;Molofsky and Swanson, 2004;Wieland et al, 2005;Eylert et al, 2010). Surprisingly, Acanthamoebae, a primary host amoeba of L. pneumophila in the aquatic environment, is auxotrophic for Arg, Ile, Leu, Met and Val (Fig.…”

Section: Synchronization Of Amino Acid Auxotrophymentioning

confidence: 99%

“…Recent evidence indicates idiosyncrasies in novel microbial strategies to trigger generation of essential nutrients from the host to power bacterial proliferation (Alkhuder et al, 2009;Price et al, 2011;Niu et al, 2012;Winter et al, 2013). Legionella pneumophila, although auxotrophic for several amino acids including cysteine, preferentially metabolizes amino acids as the main sources of energy and carbon to power intracellular replication in amoeba and human cells (Pine et al, 1979;George et al, 1980;Reeves et al, 1981;Ristroph et al, 1981;Tesh and Miller, 1981;Tesh et al, 1983;Molofsky and Swanson, 2004;Wieland et al, 2005;Eylert et al, 2010). This organism has evolved an exquisite mechanism to promote host degradation of proteins to generate a plentiful supply of amino acids to power its intracellular replication in a diverse range of amoebal hosts and the evolutionarily distant human host (Al-Khodor et al, 2008;Price et al, 2009;2010b;2011).…”

Section: Introductionmentioning

confidence: 99%

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Amoeba host‐Legionella synchronization of amino acid auxotrophy and its role in bacterial adaptation and pathogenic evolution

Price

Richards

Dwingelo

et al. 2013

Environmental Microbiology

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Summary Legionella pneumophila, the causative agent of Legionnaires’ disease, invades and proliferates within a diverse range of free-living amoeba in the environment but upon transmission to humans the bacteria hijack alveolar macrophages. Intracellular proliferation of L. pneumophila in two evolutionarily distant hosts is facilitated by bacterial exploitation of conserved host processes that are targeted by bacterial protein effectors injected into the host cell. A key aspect of microbe-host interaction is microbial extraction of nutrients from the host but understanding of this is still limited. AnkB functions as a nutritional virulence factor and promotes host proteasomal degradation of polyubiquitinated proteins generating gratuitous levels of limiting host cellular amino acids. L. pneumophila is auxotrophic for several amino acids including cysteine, which is a metabolically preferred source of carbon and energy during intracellular proliferation, but is limiting in both amoebae and humans. We propose that synchronization of bacterial amino acids auxotrophy with the host is a driving force in pathogenic evolution and nutritional adaptation of L. pneumophila and other intracellular bacteria to life within the host cell. Understanding microbial strategies of nutrient generation and acquisition in the host will provide novel antimicrobial strategies to disrupt pathogen access to essential sources of carbon and energy.

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The life stage‐specific pathometabolism of Legionella pneumophila

Eisenreich

Heuner

2016

FEBS Letters

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The genus Legionella belongs to Gram-negative bacteria found ubiquitously in aquatic habitats, where it grows in natural biofilms and replicates intracellularly in various protozoa (amoebae, ciliates). L. pneumophila is known as the causative agent of Legionnaires' disease, since it is also able to replicate in human alveolar macrophages, finally leading to inflammation of the lung and pneumonia. To withstand the degradation by its host cells, a Legionellacontaining vacuole (LCV) is established for intracellular replication, and numerous effector proteins are secreted into the host cytosol using a type four B secretion system (T4BSS). During intracellular replication, Legionella has a biphasic developmental cycle that alternates between a replicative and a transmissive form. New knowledge about the host-adapted and life stagedependent metabolism of intracellular L. pneumophila revealed a bipartite metabolic network with life stage-specific usages of amino acids (e.g. serine), carbohydrates (e.g. glucose) and glycerol as major substrates. These metabolic features are associated with the differentiation of the intracellular bacteria, and thus have an important impact on the virulence of L. pneumophila.

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Chemically defined medium for Legionella pneumophila growth

Cited by 84 publications

References 4 publications

Interaction of Bartonella henselae with endothelial cells results in rapid bacterial rRNA synthesis and replication

Interaction of Bartonella henselae with endothelial cells results in rapid bacterial rRNA synthesis and replication

Amoeba host‐Legionella synchronization of amino acid auxotrophy and its role in bacterial adaptation and pathogenic evolution

The life stage‐specific pathometabolism of Legionella pneumophila

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