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Methods for Determining Protein Translocation by the Legionella pneumophila Dot/Icm Type IV Secretion System

Legionella pneumophila the causative agent of Legionnaires’ disease, actively manipulates host cell �processes to establish a membrane-bound replication vacuole permissive for its replication. Establishment of such replication niche requires the Dot/Icm type IV secret ...

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Subcellular Localization of Legionella Dot/Icm Effectors

The translocation of effector proteins by the Dot/Icm type IV secretion system is central to the ability of Legionella pneumophila to persist and replicate within eukaryotic cells. The subcellular localization of translocated Dot/Icm proteins in host cells provides insight into th ...

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Purification and Characterization of Legionella U-Box-Type E3 Ubiquitin Ligase

Bacterial virulence proteins often mimic host eukaryotic proteins to modify or disturb host cellular �pathways. Increasing lines of evidence show that many bacterial effector proteins have E3 ubiquitin ligase activity. The effector protein LubX is one such bacterial E3 ubiquitin l ...

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Legionella Phospholipases Implicated in Infection: Determination of Enzymatic Activities

The intracellularly replicating lung pathogen Legionella pneumophila expresses a multitude of different phospholipases which are important virulence tools during host cell infection. To study the lipolytic properties including substrate specificities of potential L. ...

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The Tetrahymena and Acanthamoeba Model Systems

Although the study of protozoology has been active for centuries, very few current academic curricula incorporate requirements or even options for coursework on the study of protists; yet, protozoa are becoming widely recognized by investigators as organisms that play a significant ...

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Dictyostelium Host Response to Legionella Infection: Strategies and Assays

The professional phagocyte Dictyostelium discoideum is a simple eukaryotic microorganism, whose natural habitat is deciduous forest soil and decaying leaves, where the amoebae feed on bacteria and grow as separate, independent, single cells. In the last decade, the organism has been s ...

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The Caenorhabditis elegans Model of Legionella Infection

Caenorhabditis elegans can serve as a simple genetic host to study interactions between Legionellaceae and their hosts, and to examine the contribution of specific gene products to virulence and immunity. C. elegans nematodes have several appealing attributes as a host organism; they a ...

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Infection of Nonphagocytic Host Cells by Legionella

Legionella pneumophila is an intracellular pathogen of free-living protozoa that can also infect alveolar macrophages, L929 fibroblast cells, and HeLa cells. Infection of nonphagocytic cells by L. pneumophila can be used to study invasion mechanisms, compare infectivity of diffe ...

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The Mouse as a Model for Pulmonary Legionella Infection

Legionella pneumophila is an intracellular bacterium that was evolutionarily selected to survive in freshwater environments by infecting free-living unicellular protozoa. Once humans inhale contaminated water droplets, the bacteria reach the pulmonary alveoli where t ...

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Analyzing Caspase-1 Activation During Legionella pneumophila Infection in Macrophages

Caspase-1 is a critical factor in the innate immune response to Legionella pneumophila. The development of methods for analyzing caspase-1 activation pathways and downstream caspase-1-associated activities has helped in understanding the regulation of this protease and the si ...

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Assessment of Legionella-Specific Immunity in Mice

Legionella pneumophila is the causative agent of the potentially fatal Legionnaires’ disease in humans. Mice have proved to be valuable model organisms to study the pathogenesis of this intracellular bacterium, as well as immune responses against it. In this chapter we describe a selecti ...

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The Guinea Pig Model of Legionnaires Disease

The guinea pig pneumonia model mimics Legionnaires’ disease as seen in immunocompromised humans, with high untreated fatality rates and as such lends itself to studies of experimental chemotherapy. Guinea pig infection is also used to assess relative virulence of different Legione ...

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Human Susceptibility to Legionnaires Disease

Legionella pneumophila is a facultative intracellular pathogen that is an important cause of pneumonia. Although host factors that may predispose to acquisition of Legionnaire’s Disease (LD) include comorbid illnesses (e.g., diabetes, chronic lung disease), age, male sex, and smok ...

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cDNA Library Construction for Next-Generation Sequencing to Determine the Transcriptional Landscape of Legionella pneumophila

The adaptation of Legionella pneumophila to the different conditions it encounters in the environment and in the host is governed by a complex regulatory system. Current knowledge of these regulatory networks and the transcriptome responses of L. pneumophila is mainly based on microar ...

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Outbreak Investigations and Identification of Legionella in Contaminated Water

To avoid further cases arising from an infectious source it is essential to ensure the early identification of all potential source(s) within an identified area, or buildings, to determine if they are being managed safely; to take appropriate samples and ensure appropriate remedial actio ...

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Investigation of Outbreaks: Epidemiology

Outbreaks of Legionnaires’ disease create high levels of public anxiety and media interest and inevitably consume a great deal of public health resources. Investigations should begin as early as possible in order to rapidly identify suspected sources of infection, control the outbre ...

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Sequence-Based Identification of Legionella

Legionella strains are considered biologically inert with respect to traditional identification schemes. Various phenotypic alternatives have been attempted but all have lacked resolution as additional species have been added to what is proving to be a large genus. Only sequence ...

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Identification of Legionella in Clinical Samples

Currently, several methods are used for the detection of Legionella in clinical samples, and these methods constitute part of the criteria for defining legionellosis cases. Urinary antigen detection is the first-line diagnostic test, although this test is limited to L. pneumophila ser ...

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Identification of Legionella in the Environment

Legionella is ubiquitous in freshwater systems worldwide and can also be found in soil. Legionellosis may be caused by inhalation of aerosolized water or soil particles containing Legionella. Isolation of Legionella from the environment is an essential step in outbreak investigati ...

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Typing Methods for Legionella

In this chapter we describe the methods currently used for subgrouping Legionella pneumophila and other non-pneumophila species. In the first part we describe monoclonal antibody (mAb) subgrouping, either by indirect immunofluorescence or indirect ELISA methods. These monoc ...

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