Characterization of a Lipoprotein Common to
            <i>Legionella</i>
            Species as a Urinary Broad-Spectrum Antigen for Diagnosis of Legionnaires' Disease

Park, Dae Won; Park, Seung Chul; Chun, Byung Chul

doi:10.1128/jcm.41.7.2974-2979.2003

Cited by 35 publications

(21 citation statements)

References 27 publications

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“…The UAG test was the most commonly used test with the highest sensitivity and lowest cost, but performance may be improved further by coverage of more Legionella serogroups and species (25). Although culture is recommended for detecting strains that would be missed by an Lp1-specific antigen and an L. pneumophila PCR, it is technically demanding with low sensitivity.…”

Section: Discussionmentioning

confidence: 99%

Utility of PCR, Culture, and Antigen Detection Methods for Diagnosis of Legionellosis

et al. 2015

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The goal of this retrospective study was to evaluate the performance of different diagnostic tests for Legionnaires' disease in a clinical setting where Legionella pneumophila PCR had been introduced. Electronic medical records at the Cleveland Clinic were searched for Legionella urinary antigen (UAG), culture, and PCR tests ordered from March 2010 through December 2013. For cases where two or more test methods were performed and at least one was positive, the medical record was reviewed for relevant clinical and epidemiologic factors. Excluding repeat testing on a given patient, 19,912 tests were ordered (12,569 UAG, 3,747 cultures, and 3,596 PCR) with 378 positive results. The positivity rate for each method was 0.4% for culture, 0.8% for PCR, and 2.7% for UAG. For 37 patients, at least two test methods were performed with at least one positive result: 10 (27%) cases were positive by all three methods, 16 (43%) were positive by two methods, and 11 (30%) were positive by one method only. For the 32 patients with medical records available, clinical presentation was consistent with proven or probable Legionella infection in 84% of the cases. For those cases, the sensitivities of culture, PCR, and UAG were 50%, 92%, and 96%, respectively. The specificities were 100% for culture and 99.9% for PCR and UAG. L egionnaires' disease is an atypical, respiratory illness associated with exposure to water colonized with Legionella species (1). In the United States, up to 18,000 hospitalizations occur each year for legionellosis, with the vast majority (70% to 92%) attributed to Legionella pneumophila serogroup 1 (Lp1) (2). This predominance of Lp1 disease is thought to be a reflection of virulence rather than environmental distribution (3-5). Besides Lp1, the strains most commonly associated with human disease are other L. pneumophila serogroups, Legionella micdadei, Legionella bozemanii, and Legionella longbeachae (2, 6). Risk factors for legionellosis include whirlpool spa exposure, recent overnight travel or plumbing repairs (two weeks prior to onset of symptoms), immunosuppression, alcoholism, diabetes, malignancy, hepatic or renal failure, chronic obstructive lung disease, smoking history, and patient age of Ͼ50 years (7). Patients with Legionnaires' disease often require intensive care unit (ICU) admission, have failed outpatient antimicrobial treatment, or may meet criteria for nosocomial pneumonia (8).The urinary antigen (UAG) test is commonly used to diagnose Legionnaires' disease because sputum production is limited and Legionella culture requires special techniques. The Lp1 antigen is typically detectable in urine beginning 2 to 3 days after the onset of clinical symptoms until 2 months after clearance of disease but may persist for a much longer period of time (2). The widespread availability of rapid, FDA-cleared, Lp1 UAG tests in the United States coincided with a 76% decrease in mortality rate (34% to 8%) from Legionnaires' disease during 1985 to 2009 (9, 10). However, it has been suggested that non-Lp1 Legio...

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Section: Discussionmentioning

confidence: 99%

Utility of PCR, Culture, and Antigen Detection Methods for Diagnosis of Legionellosis

et al. 2015

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“…Although Legionella species-common PAL antigen has been characterized as the most abundant protein of the soluble fractions (Kim et al, 2003), its role in pathogenesis is unknown. In the present study, in vitro rPAL-stimulated macrophages induced effectively IL-6 and TNF-α production in dose-dependent fashion, mediated by TLR2 signaling.…”

Section: Discussionmentioning

confidence: 99%

“…The protein concentration was determined using the Bio-Rad protein assay kit (Bio-Rad Laboratories, Hercules, CA). The protein was identified by SDS-PAGE and immunoblotting with rabbit anti-rPAL IgG (Kim et al, 2003) (data not shown). To eliminate contaminating LPS from the rPAL preparation, all reagents were prepared with endotoxin-free solutions.…”

Section: Production and Purification Of A Recombinant Lipoproteinmentioning

confidence: 99%

“…Our previous study first demonstrated that rPAL of L. pneumophila, as a vaccine candidate, was a potent activator of PAL-specific B and T cell responses in BALB/c mice immunized with either the rPAL or plasmid DNA PAL (Yoon et al, 2002). Subsequently, the PAL was characterized as a soluble antigen excreted from urine of patients with Legionella pneumonia (Kim et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

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Legionella lipoprotein activates toll-like receptor 2 and induces cytokine production and expression of costimulatory molecules in peritoneal macrophages

Shim

Kim

et al. 2009

Exp Mol Med

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Legionella bacterium, an intracellular pathogen of mononuclear phagocytes, causes acute fatal pneumonia, especially in patients with impaired cellular immune responses. Until recently, however, the toll-like receptor (TLR) engagement of bacterial proteins derived from Legionella is uncertain. We previously showed that a 19-kDa highly conserved peptidoglycan-associated lipoprotein (PAL) of Legionella pneumophila induced the PAL-specific B cell and T cell responses in mice. In this study, we observed that the rPAL antigen of L. pneumophila, as an effector molecule, activated murine macrophages via TLR2 and produced proinflammatory cytokines such as IL-6 and TNF-α. In both BALB/c and TLR4-deficient C3H/HeJ mice, pretreatment of macrophages with anti-TLR2 mAb showed severely impaired cytokine production in response to the rPAL. In addition, in vitro the rPAL treatment increased the cell surface expression of CD40, CD80, CD86 and MHC I/II molecules. We further showed that the synthetic CpG-oligodeoxynucleotides (CpG ODN) coadministered with the rPAL enhanced IL-12 and IL-6 production and expression of CD40, CD80 and MHC II compared to the rPAL treatment alone. In conclusions, these results indicate that Legionella PAL might activate macrophages via a TLR2-dependent mechanism which thus induce cytokine production and expression of costimulatory and MHC molecules.

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“…In E. coli, Pal contributes to the maintenance of cell wall integrity (22) and causes inflammation after release into the bloodstream in gram-negative sepsis (12). The expression of Pal is required for virulence in experimental Haemophilus ducreyi infection (8), and Legionella pneumophila Pal has been proposed as a possible diagnostic antigen for Legionnaires' disease (18). The identity of the other LMW protein, which for unknown reasons was not identified by Western blotting of the two-dimensional gel, is yet to be determined.…”

Section: Discussionmentioning

confidence: 99%

Suppression of Serum Antibody Responses by Pertussis Toxin after Respiratory Tract Colonization byBordetella pertussisand Identification of an Immunodominant Lipoprotein

Carbonetti¹,

Артамонова²,

Andreasen³

et al. 2004

Infect Immun

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Pertussis toxin (PT), a virulence factor secreted by Bordetella pertussis, contributes to respiratory tract infection and disease caused by this pathogen. By comparing a wild-type (WT) B. pertussis strain to a mutant strain with an in-frame deletion of the ptx genes encoding PT (⌬PT), we recently found that the lack of PT confers a significant defect in respiratory tract colonization in mice after intranasal inoculation. In this study, we analyzed serum antibody responses in mice infected with the WT or ⌬PT strain and found that infection with the ⌬PT strain elicited greater responses to several B. pertussis antigens than did infection with the WT, despite the lower colonization level achieved by the ⌬PT strain. The same enhanced antibody response was observed after infection with a strain expressing an enzymatically inactive PT; but this response was not observed after infection with B. pertussis mutant strains lacking filamentous hemagglutinin or adenylate cyclase toxin, nor when purified PT was administered with the ⌬PT inoculum, indicating a specific role for PT activity in this immunosuppressive effect. In particular, there were consistent strong serum antibody responses to one or more low-molecular-weight antigens after infection with the ⌬PT strain. These antigens were Bvg independent, membrane localized, and also expressed by the closely related pathogens Bordetella parapertussis and Bordetella bronchiseptica. Two-dimensional gel electrophoresis and mass spectrometry were used to identify one of the immunodominant low-molecular-weight antigens as a protein with significant sequence homology to peptidoglycan-associated lipoprotein in several other gram-negative bacterial species. However, a serum antibody response to this protein alone did not protect mice against respiratory tract infection by B. pertussis.Pertussis toxin (PT) is an important virulence factor produced exclusively by Bordetella pertussis, a gram-negative bacterial pathogen that colonizes the human respiratory tract and causes a disease known as whooping cough or pertussis. PT is a member of the AB 5 structural class of bacterial toxins (41, 43), consisting of an enzymatically active A subunit (S1) that ADP-ribosylates the alpha subunit of the Gi family of heterotrimeric G proteins in mammalian cells (17, 30) and a pentameric B oligomer that binds unidentified glycoconjugate receptors on cells (5, 48). PT activity has a wide range of effects on signaling pathways and normal function in mammalian cells (35,49). The administration of purified PT to experimental animals can reproduce almost all of the systemic symptoms associated with human pertussis disease, such as histamine sensitivity, lymphocytosis, and insulinemia (29, 31, 32), but its role in respiratory tract colonization and disease is uncertain. Several studies of mice intranasally infected with bacteria have shown that PT plays a role in the overall respiratory tract infection by B. pertussis (2,10,45,46). Recently, it was found that PT is a significant colonization factor for resp...

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Characterization of a Lipoprotein Common to Legionella Species as a Urinary Broad-Spectrum Antigen for Diagnosis of Legionnaires' Disease

Cited by 35 publications

References 27 publications

Utility of PCR, Culture, and Antigen Detection Methods for Diagnosis of Legionellosis

Utility of PCR, Culture, and Antigen Detection Methods for Diagnosis of Legionellosis

Legionella lipoprotein activates toll-like receptor 2 and induces cytokine production and expression of costimulatory molecules in peritoneal macrophages

Suppression of Serum Antibody Responses by Pertussis Toxin after Respiratory Tract Colonization byBordetella pertussisand Identification of an Immunodominant Lipoprotein

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