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PubMed2004

Caenorhabditis elegans-based screen identifies Salmonella virulence factors required for conserved host-pathogen interactions.

A Caenorhabditis elegans-Salmonella enterica host-pathogen model was used to identify both novel and previously known S. enterica virulence factors (HilA, HilD, InvH, SptP, RhuM, Spi4-F, PipA, VsdA, RepC, Sb25, RfaL, GmhA, LeuO, CstA, and RecC), including several related to the type III secretion system (TTSS) encoded in Salmonella pathogenicity island 1 (SPI-1). Mutants corresponding to presumptive novel virulence-related genes exhibited diminished ability to invade epithelial cells and/or to induce polymorphonuclear leukocyte migration in a tissue culture model of mammalian enteropathogenesis. When expressed in C. elegans intestinal cells, the S. enterica TTSS-exported effector protein SptP inhibited a conserved p38 MAPK signaling pathway and suppressed the diminished pathogenicity phenotype of an S. enterica sptP mutant. These results show that C. elegans is an attractive model to study the interaction between Salmonella effector proteins and components of the innate immune response, in part because there is a remarkable overlap between Salmonella virulence factors required for human and nematode pathogenesis.

Animals↗
PubMed2006

In vitro inhibitory effects of Polygonum cuspidatum on bacterial viability and virulence factors of Streptococcus mutans and Streptococcus sobrinus.

OBJECTIVES: Polygonum cuspidatum has been used in Korean folk medicine to improve oral hygiene. This study was performed to evaluate the effects of methanol extract from root of P. cuspidatum (MEP) on bacterial viability and the virulence factors of Streptococcus mutans and Streptococcus sobrinus. METHODS: To test the effects of MEP on bacterial viability, we determined the minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) against 20 bacterial strains, including S. mutans and S. sobrinus, using a micro-dilution assay. In case of S. mutans and S. sobrinus, the assays for time-kill and bacterial growth rate at sub-MIC concentrations were also performed. To determine effects of the extract on the virulence factors of S. mutans and S. sobrinus, the assays for sucrose-dependent adherence, water-insoluble glucan formation, glycolytic acid production, and acid tolerance were performed at sub-MIC levels. Phytochemical analysis for constituents of MEP was carried out. RESULTS: MEP showed a broad antibacterial range (MIC 0.5-4 mg/ml). The MBC was two to four times higher than the MIC. The time-kill curves showed S. mutans and S. sobrinus were significantly killed after 1h of incubation. At sub-MIC levels, doubling times of S. mutans and S. sobrinus dose-dependently increased up to 211% and 123%, respectively. At sub-MIC levels, MEP also showed inhibitory effects on the virulence factors of S. mutans and S. sobrinus in a dose-dependent fashion. Phytochemical analysis revealed the presence of alkaloids, sterol/terpenes, tannins, flavonoids, and carbohydrates. CONCLUSION: These data indicate that MEP has inhibitory effects on bacterial viability at higher concentrations (> or =MIC) and the virulence factors of S. mutans and S. sobrinus at sub-MIC concentrations, suggesting that it might be useful for the control of dental plaque formation and subsequent dental caries formation.

Anti-Bacterial Agents↗
PubMed2006

Identification and characterization of a novel bacterial virulence factor that shares homology with mammalian Toll/interleukin-1 receptor family proteins.

Many important bacterial virulence factors act as mimics of mammalian proteins to subvert normal host cell processes. To identify bacterial protein mimics of components of the innate immune signaling pathway, we searched the bacterial genome database for proteins with homology to the Toll/interleukin-1 receptor (TIR) domain of the mammalian Toll-like receptors (TLRs) and their adaptor proteins. A previously uncharacterized gene, which we have named tlpA (for TIR-like protein A), was identified in the Salmonella enterica serovar Enteritidis genome that is predicted to encode a protein resembling mammalian TIR domains, We show that overexpression of TlpA in mammalian cells suppresses the ability of mammalian TIR-containing proteins TLR4, IL-1 receptor, and MyD88 to induce the transactivation and DNA-binding activities of NF-kappaB, a downstream target of the TIR signaling pathway. In addition, TlpA mimics the previously characterized Salmonella virulence factor SipB in its ability to induce activation of caspase-1 in a mammalian cell transfection model. Disruption of the chromosomal tlpA gene rendered a virulent serovar Enteritidis strain defective in intracellular survival and IL-1beta secretion in a cell culture infection model using human THP1 macrophages. Bacteria with disrupted tlpA also displayed reduced lethality in mice, further confirming an important role for this factor in pathogenesis. Taken together, our findings demonstrate that the bacterial TIR-like protein TlpA is a novel prokaryotic modulator of NF-kappaB activity and IL-1beta secretion that contributes to serovar Enteritidis virulence.

Amino Acid Sequence↗
PubMed1993

Environmental regulation of virulence factors in Bordetella species.

Many bacteria respond in a coordinate manner to environmental changes. External stimuli, sensed by receptors, are transduced to regulatory proteins which participate in well defined pathways of gene expression by varying their structure and mode of action. The network of environmental signal transduction is responsible for a fine and continuous communication between the host and the pathogenic bacteria. As a result, the gene expression machinery of the pathogen is modified continuously, in order to establish the optimal conditions for bacterial survival and multiplication.

Amino Acid Sequence↗
PubMed2001

[Virulence factors associated with E. coli neonatal meningitis].

Escherichia coli K1 is the leading cause of gram-negative bacterial meningitis in neonates. It is associated with a mortality rate as high as 40%, and more than half of the survivors have neurologic sequelae. Bacterial meningitis is the result of bacterial translocation from gastrointestinal tract to the blood and from blood to the central nervous system. Successful crossing of the BBB by E. coli K1 requires (a) a high degree of bacteremia and (b) several E. coli determinants contributing to invasion of BMEC such as the K1 capsule, Sfa, Ibe proteins, and CNF1. A better understanding for the molecular basis of E. coli K1 penetration of the BBB could potentially lead to the development of novel therapeutic and preventative strategies for E. coli K1 meningitis.

Blood-Brain Barrier↗
PubMed2005

Staphylococcus aureus leucocidin, a virulence factor in bovine mastitis.

The involvement of Staphylococcus aureus exosecretions in bovine udder infection (Younis et al. 2003) suggests that four different monomer protein bands appearing between 36 and 31 kDa, are associated with the severity of the cow's infection response. Three out of these four bands have been identified by means of protein sequencing. Band B, with a MW of 35 kDa was identified as Panton-Valentaine leucocidin LukF'-PV chain- Staph. aureus; band C, with a MW of 32 kDa was identified as leucocidin chain LukM precursor- Staph. aureus; and band D was found to be similar, but not identical, to phosphatidylinositol-specific phospholipase-C-X. Bands B and C were purified by gel filtration using FPLC. The ability of these proteins to induce udder inflammation in vivo, and proliferation response in vitro and cytokine secretion were tested for both the crude exosecretions and purified bands. Three cows were inoculated intracisternally, with three quarters receiving either 0.007-0.008 mg (as total proteins) of Staph. aureus FR2449/1 bacterial exosecretion, pooled fraction 39-41 (bands B and C), or culture broth medium. The fourth quarter was left free as a control. Quarters that received fraction 39-41 of Staph. aureus FR2449/1, exhibited induced inflammation, which was indicated by increased somatic cell count and enhanced NAGase activity that was significantly higher than that of the original Staph. aureus FR2449/1 bacterial exosecretion. Proliferation tests of bovine blood lymphocytes in vitro showed that the pooled fraction 39-41 stimulated bovine proliferation of mononuclear cells much more than the original Staph. aureus FR2449/1 bacterial exosecretion. Secretion of TNF-alpha, IL-1beta, IL-6 and IL-8 was in accordance with the contents of LukF'-PV and LukM precursor in the exosecretions. The results suggest that LukM/ LukF' induce inflammation into the udder by a mechanism similar to that of LPS or by a unique mechanism(s) which requires further investigation.

Amino Acid Sequence↗
PubMed1996

Known bacterial virulence factors do not explain the variation in urinary cytokine levels in patients with urosepsis.

We measured urinary endotoxin, IL-6 and IL-8 levels in 23 patients with gram-negative urosepsis. The endotoxin and cytokine levels showed a 100-1000 fold range. No correlation was found between levels of urinary endotoxin, and IL-6 or IL-8 levels. In all cases bacterial numbers were > or = 10(5) CFU ml-1 urine. The endotoxin content of the isolated microorganisms neither correlated with the urinary cytokine levels, nor with IL-6 and IL-8 levels obtained in vitro when 10(3) log-phase CFU of each of the bacteria were incubated with heparinized whole blood of three healthy donors. Neither the haemolysin phenotype of the bacteria, nor the presence of the P-pili gene was correlated with the cytokine response in vivo or in vitro. Other factors than known bacterial virulence factors apparently contribute to the wide variation in urinary cytokine levels in urinary tract infection.

Cytokines↗
PubMed2004

Temporal shedding patterns and virulence factors of Escherichia coli serogroups O26, O103, O111, O145, and O157 in a cohort of beef calves and their dams.

This study investigated the shedding of Escherichia coli O26, O103, O111, O145, and O157 in a cohort of beef calves from birth over a 5-month period and assessed the relationship between shedding in calves and shedding in their dams, the relationship between shedding and scouring in calves, and the effect of housing on shedding in calves. Fecal samples were tested by immunomagnetic separation and by PCR and DNA hybridization assays. E. coli O26 was shed by 94% of calves. Over 90% of E. coli O26 isolates carried the vtx(1), eae, and ehl genes, 6.5% carried vtx(1) and vtx(2), and one isolate carried vtx(2) only. Serogroup O26 isolates comprised seven pulsed-field gel electrophoresis (PFGE) patterns but were dominated by one pattern which represented 85.7% of isolates. E. coli O103 was shed by 51% of calves. Forty-eight percent of E. coli O103 isolates carried eae and ehl, 2% carried vtx(2), and none carried vtx(1). Serogroup O103 isolates comprised 10 PFGE patterns and were dominated by two patterns representing 62.5% of isolates. Shedding of E. coli O145 and O157 was rare. All serogroup O145 isolates carried eae, but none carried vtx(1) or vtx(2). All but one serogroup O157 isolate carried vtx(2), eae, and ehl. E. coli O111 was not detected. In most calves, the temporal pattern of E. coli O26 and O103 shedding was random. E. coli O26 was detected in three times as many samples as E. coli O103, and the rate at which calves began shedding E. coli O26 for the first time was five times greater than that for E. coli O103. For E. coli O26, O103, and O157, there was no association between shedding by calves and shedding by dams within 1 week of birth. For E. coli O26 and O103, there was no association between shedding and scouring, and there was no significant change in shedding following housing.

Animal Husbandry↗
PubMed1995

Virulence factors in the colonization and persistence of bacteria in the airways.

Haemophilus influenzae, Streptococcus pneumoniae, Moraxella catarrhalis, Staphylococcus aureus, Klebsiella pneumoniae, and Pseudomonas aeruginosa are commonly isolated from sputum specimens of patients with lower respiratory tract infections. Haemophilus influenzae, S. pneumoniae, and M. catarrhalis have several pathogenic properties in common. These bacteria are able to interact with mucus, to exert ciliotoxic activity, to adhere to bronchial epithelial cells, and to invade airway epithelium. Haemophilus influenzae and S. pneumoniae strains persist for many months in the respiratory tract of patients with chronic obstructive pulmonary disease (COPD), despite the specific antibodies present in serum and sputum against the persistent strain. Especially during exacerbations persistent strains with changes in their antigenic composition are isolated. Among H. influenzae strains, the antigenic characteristics of the outer membrane protein composition vary. Variation in S. pneumoniae occurs in capsular polysaccharides, the major immunogens of this bacterium. Such variations affect the efficacy of the antibody-mediated defense mechanisms against the bacteria. Between exacerbations, particularly H. influenzae, S. pneumoniae strains are recovered from the sputum of patients with COPD. Recovery may continue for periods up to 2 yr, although not continuously. Besides ineffective antibody-mediated defense mechanisms, it is likely that hiding of the bacteria in tissue contributes to the persistence of these bacteria in patients with COPD.

Bacterial Infections↗
PubMed1991

[Adhesiveness in Neisseria meningitidis. A virulence factor?].

The role of adhesivity as a factor of virulence in cases of N. meningitidis has not yet been established. Unlike other bacteria in which the presence of adhesivity reflects an enhanced virulence, a high capacity of adherence is associated with a low virulence and invasive capacity in cases of meningococcus. In order to gain more insight into the knowledge of this phenomenon we have studied the adhesivity of 109 strains of N. meningitidis to human pharyngeal epithelial cells. Twenty nine out of the 109 strains were isolated from blood or spinal fluid of patients with meningococcal infection (meningitis or septicemia), whereas the remaining 80 strains were obtained from pharyngeal smears of healthy carriers. The adhesivity was measured as the number of meningococci adhered to 50 epithelial cells according to Craven's scale. Strains of healthy carriers showed a greater adhesivity than that of patients (p less than 0.001). The relevance of the pharyngeal area in the evaluation of the adhesive capacity is in accordance with the hypothesis that meningococcal adhesivity decreases when the microorganism crosses the pharyngeal epithelium. The results would support the concept that the virulence of N. meningitidis is related to its adhesiveness.

Bacterial Adhesion↗
PubMedDate not supplied

[The detection of the Shiga toxin antigen in connection with other virulence factors--the O and K antigens of enterobacteria].

The results of the detection of Shiga toxin antigen in culture filtrates of 73 enterobacterial strains by the methods of countercurrent electrophoresis and coagglutination, as well as their correlation with the presence of other factors of virulence, O- and K-antigens, have been analyzed. Sharply pronounced inverse correlation between the capacity of enterobacteria to synthesize Shiga (Shiga-like) toxins and their capacity to synthesize acidic polysaccharide surface-specific K- (Vi-) antigens has been established.

Agglutination Tests↗
PubMed1995

[Bacterial virulence factors and their role in the pathogenesis of pyogenic infections of the abdominal cavity and mediastinum].

Purulent peritonitis are caused predominantly by anaerobic bacteria which come from physiological intestinal flora. Mediastinitis is caused amidst other etiologic factors also by bacteria inhabiting the oropharyngeal region, as well as microorganisms causing diseases localized in the proximity of mediastinum. Anaerobic sporulating bacteria both Gram positive and negative cause often miscellaneous infections due to Staphylococcus aureus, Enterobacteriacae, Streptococcus, Enterococcus. It involves a group of bacteria which are able to produce a fibrin network in their proximity, to resist phagocytosis by their structures, to distruct the components of the complement system and immunoglobulins, to impair membranes of cells which leads by means of their factors of virulence to formation of defectuous immunity. Microbiological examination requires the material to be sent for anaerobic cultivation and the antimicrobial therapy must take into account its polymicrobic etiology. (Ref. 23.)

Abdomen↗
PubMed2003

Priming virulence factors for delivery into the host.

Several medically important Gram-negative bacterial pathogens inject virulence factors into host cells through a type III secretion system and specialized bacterial chaperones are required for their effective delivery. Recent structural work shows that these chaperones maintain virulence factors in a partially non-globular conformation that is primed for unfolding and translocation through the 'injectisome'.

Animals↗
PubMed1996

Virulence factors of medically important fungi.

Human fungal pathogens have become an increasingly important medical problem with the explosion in the number of immunocompromised patients as a result of cancer, steroid therapy, chemotherapy, and AIDS. Additionally, the globalization of travel and expansion of humankind into previously undisturbed habitats have led to the reemergence of old fungi and new exposure to previously undescribed fungi. Until recently, relatively little was known about virulence factors for the medically important fungi. With the advent of molecular genetics, rapid progress has now been made in understanding the basis of pathogenicity for organisms such as Aspergillus species and Cryptococcus neoformans. The twin technologies of genetic transformation and "knockout" deletion construction allowed for genetic tests of virulence factors in these organisms. Such knowledge will prove invaluable for the rational design of antifungal therapies. Putative virulence factors and attributes are reviewed for Aspergillus species, C. neoformans, the dimorphic fungal pathogens, and others, with a focus upon a molecular genetic approach. Candida species are excluded from coverage, having been the subject of numerous recent reviews. This growing body of knowledge about fungal pathogens and their virulence factors will significantly aid efforts to treat the serious diseases they cause.

Antigens, Fungal↗
PubMed2006

[The role of the quorum sensing system in a rat model of Pseudomonas aeruginosa pulmonary infection and its relationship with the expression of virulence factors].

OBJECTIVE: A variety of in vitro experiments indicated that most of the extracellular virulence factors of Pseudomonas aeruginosa (PA) were controlled by the quorum sensing system. The aim of this study was to explore the role of the quorum sensing system in a rat model of PA pulmonary infection. METHODS: Rats were infected intratracheally with PA strain PAO1 or PAO1-JP2 (lasI rhlI double mutant) which was embedded in seaweed alginate bead. The difference of bacteriology and pathology between the two groups was compared. The in vitro activity of elastase was measured by the Congo Red method and the expression of exotoxin A was detected by Western blot. RESULTS: At the 14th and 28th days after challenge, the bacterial colony counts in the PAO1-JP2 group were (9.6 +/- 3.3) lgCFU/g and (4.2 +/- 3.1) lgCFU/g respectively; however, those of the PAO1 group were (11.3 +/- 2.8) lgCFU/g and (9.1 +/- 1.5) lgCFU/g respectively, which showed significant difference (P < 0.05). The lung index of macroscopic pathology (LIMP) and the score of macroscopic pathology of the PAO1-JP2 group were significantly lower than those of the PAO1 group at 7th, 14th and 28th days after challenge. The size of lung abscesses or granuloma in the lung under the microscope and the score of microscopic pathology of the PAO1-JP2 group were also significantly lower at the 7th, 14th and 28th days. The in vitro activity of elastase of the PAO1-JP2 strain was 0.02 +/- 0.00 and that of the PAO1 strain was 0.35 +/- 0.03, which showed significant difference (P < 0.05). The expression of exotoxin A could be detected by Western blot in the PAO1 strain but not in the PAO1-JP2 strain. CONCLUSION: When the quorum sensing system of PA was deficient the expression of some virulence factors, such as elastase and exotoxin A, were decreased, which resulted in milder pulmonary infection.

Animals↗
PubMed2002

Common themes among bacteriophage-encoded virulence factors and diversity among the bacteriophages involved.

There are common themes among bacteriophage-encoded virulence factors, which include the well-characterized bacterial toxins and proteins that alter antigenicity as well as several new classes of bacteriophage-encoded proteins such as superantigens, effectors translocated by a type III secretion system, and proteins required for intracellular survival and host cell attachment. These virulence factors are encoded by a diversity of bacteriophages, members of the viral families Siphoviridae, Podoviridae, Myoviridae and Inoviridae, with some bacteriophages having characteristics of more than one virus family. The location of virulence genes within the bacteriophage genomes is non-random and consistent with an origin via imprecise prophage excision or as either transferable cassettes or integral components of the bacteriophage genome.

Bacterial Adhesion↗
PubMed2006

Inhibition of swarming and virulence factor expression in Proteus mirabilis by resveratrol.

Resveratrol (3,5,4-trihydroxy-trans-stilbene) is a phytoalexin compound with anti-inflammatory and antioxidant activities. The effect of resveratrol on swarming and virulence factor expression of Proteus mirabilis, an important pathogen infecting the urinary tract, was determined on swarming agar plates with and without the compound. Bacteria harvested at different times were assayed for cell length and the production of flagella, haemolysin and urease. Resveratrol inhibited P. mirabilis swarming and virulence factor expression in a dose-dependent manner. Resveratrol significantly inhibited swarming at 15 microg ml(-1), and completely inhibited swarming at 60 microg ml(-1). Inhibition of swarming and virulence factor expression was mediated through RsbA, a His-containing phosphotransmitter of the bacterial two-component signalling system possibly involved in quorum sensing. Complementation of an rsbA-defective mutant with the rsbA gene restored its responsiveness to resveratrol. The compound also inhibited the ability of P. mirabilis to invade human urothelial cells. These findings suggest that resveratrol has potential to be developed as an antimicrobial agent against P. mirabilis infection.

Anti-Bacterial Agents↗
PubMed2002

Sequence analysis of the genes encoding for the major virulence factors of Bacillus anthracis vaccine strain 'Carbosap'.

AIMS: This study was performed to analyse the molecular characteristics of genes encoding for the major virulence factors in Bacillus anthracis vaccine strain 'Carbosap' compared with the wild B. anthracis strain, to evaluate the basis of attenuation. METHODS AND RESULTS: The molecular characteristics of the B. anthracis 'Carbosap' vaccine strain, used as vaccine in Italy, were analysed in comparison with a B. anthracis virulent strain. Despite the presence of the two virulence plasmids pXO1 and pXO2, the 'Carbosap' strain proved to be protective for cattle. The presence of the regulatory genes atxA and pagR and the gerX operon, known to be involved in the virulence, was verified. In addition, all genes were sequenced. The results showed that no molecular differences between 'Carbosap' and the virulent strain were evident. CONCLUSIONS: The results of this study indicate that the attenuation of the 'Carbosap' vaccine strain is not due to the lack of virulence genes or to modifications occurring on the sequence of these genes. Therefore, other virulence factors, still unknown, could be involved in the pathogenic mechanisms. SIGNIFICANCE AND IMPACT OF THE STUDY: This paper adds new information regarding the molecular characteristics of the vaccine strain 'Carbosap' and highlights the need to better understand the virulence factors involved in the pathogenicity of B. anthracis strains.

Anthrax↗