Role of swarming migration in the pathogenesis of bacillus endophthalmitis.

PURPOSE. Bacillus cereus causes one of the most rapidly blinding forms of bacterial endophthalmitis. Migration of B. cereus throughout the eye during endophthalmitis is a unique aspect of this disease that may contribute to intraocular virulence. This study was conducted to analyze the contribution of swarming and intraocular migration to the pathogenesis of experimental endophthalmitis. METHODS. Eyes were injected intravitreally with 100 colonyforming units (CFU) of either wild-type, nonswarming, or swarming-complemented strains of B. cereus. Pathogenicity was compared throughout the course of infection by biomicroscopy, histology, electroretinography, and bacterial and inflammatory cell quantitation. RESULTS. Wild-type, nonswarming, and swarming-complemented B. cereus strains grew to a similar number in the vitreous throughout the course of infection. Unlike the wild-type and swarming-complemented strains, the nonswarming mutant did not migrate to the anterior segment during infection. The rate of decrease in retinal responses of eyes infected with the all strains was similar, resulting in near complete elimination of retinal function by 12 hours. All Bacillus strains caused similar degrees of posterior segment inflammation and retinal destruction. However, the accumulation of inflammatory cells in the anterior chamber, hyphemae, and corneal ring abscesses did not occur in eyes infected with the nonswarming mutant. CONCLUSIONS. The deficiency in swarming had little effect on retinal function loss or the overall course or severity of experimental B. cereus endophthalmitis. However, a deficiency in swarming prevented Bacillus from migrating to the anterior segment, leading to less severe anterior segment disease

Genotypic and phenotypic properties of Candida parapsilosis sensu strictu strains isolated from different geographic regions and body sites

<p>Abstract</p> <p>Background</p> <p><it>Candida parapsilosis </it>is known to show limited genetic variability, despite different karyotypes and phenotypes have been described. To further investigate this aspect, a collection of 62 <it>sensu strictu </it><it>C. parapsilosis </it>independent isolates from 4 geographic regions (Italy, n = 19; New Zealand, n = 15; Argentina, n = 14; and Hungary, n = 14) and different body sites (superficial and deep seated) were analysed for their genetic and phenotypic traits. Amplification fragment length polymorphism (AFLP) analysis was used to confirm species identification and to evaluate intraspecific genetic variability. Phenotypic characterisation included clinically relevant traits, such as drug susceptibility, in vitro biofilm formation and aspartyl protease secretion.</p> <p>Results</p> <p>AFLP genotyping showed little variation among isolates, when the presence/absence of bands was considered. However, when AFLP profiles were compared by relative intensity for each fragment, a significant level of variation and geographical clustering was observed. All isolates were found to be susceptible to commonly used antifungals, although a reduced susceptibility to echinocandins was observed in all isolates. <it>C. parapsilosis </it>isolates from different geographic origins varied in the number of biofilm producers, with a higher prevalence of producers isolated in Hungary and Argentina. The frequency of secreted proteinase producers also varied in isolates obtained from different areas, with a higher number of proteinase producers found in Italy and New Zealand. Interestingly, biofilm production and proteinase secretion were negatively correlated. This finding could be explained by assuming that proteinase activity plays a role in detachment and release from a mature biofilm, via degradation of <it>C. parapsilosis </it>adhesins and/or extracellular matrix components, as observed for other microorganisms.</p> <p>Conclusions</p> <p>The low number of polymorphic AFLP bands (18 out of 80) obtained for <it>C. parapsilosis </it>isolates is in agreement with the limited sequence variability described for this species. However, when band intensity was included in the analysis, geographical clustering was observed. Expression of virulence factors varied among strains isolated from different geographical regions, with biofilm and proteinase producers more frequently isolated from Hungary and Italy, respectively.</p

Identification of distinct lymphocyte subsets responding to subcellular fractions of Mycobacterium bovis bacille calmette-Guerin (BCG)

In order to investigate the ability of Mycobacterium bovis BCG vaccination to induce immune responses toward different classes of mycobacterial antigens and the cell populations involved in such responses, proliferation of distinct human lymphocyte subsets from BCG-vaccinated donors in response to different subcellular fractions of BCG was analysed and compared with that of not sensitized subjects. Proliferation of different cell subsets was evaluated by flow cytometric determination of bromodeoxyuridine incorporated into DNA of dividing cells and simultaneous identification of cell surface markers. Although a certain degree of variability was observed among different donors, after 6 days of in vitro stimulation BCG-vaccinated subjects displayed, as a mean, a stronger blastogenic response to all the classes of antigens compared with non-sensitized ones. PPD, culture filtrates and membrane antigens induced a predominant proliferation of CD4(+) T cells. In contrast, preparations enriched in cytosolic antigens elicited strong proliferation of gamma delta(+) T cells which, as a mean, represented 55% of the proliferating cells. Although to a lesser extent, proliferation of gamma delta(+) T cells was also elicited by preparations enriched in membrane and cell wall antigens. In response to the latter preparation proliferation of CD4(+) T cells and CD16(+)/CD3(-) (natural killer (NK)) cells was observed, as well. In particular, cell wall antigens were found to induce significantly higher levels of proliferation of NK cells compared with all the other classes of antigens

FlhF Is Required for Swarming Motility and Full Pathogenicity of Bacillus cereus

Besides sporulation, Bacillus cereus can undergo a differentiation process in which short swimmer cells become elongated and hyperflagellated swarmer cells that favor migration of the bacterial community on a surface. The functionally enigmatic flagellar protein FlhF, which is the third paralog of the signal recognition particle (SRP) GTPases Ffh and FtsY, is required for swarming in many bacteria. Previous data showed that FlhF is involved in the control of the number and positioning of flagella in B. cereus. In this study, in silico analysis of B. cereus FlhF revealed that this protein presents conserved domains that are typical of SRPs in many organisms and a peculiar N-terminal basic domain. By proteomic analysis, a significant effect of FlhF depletion on the amount of secreted proteins was found with some proteins increased (e.g., B component of the non-hemolytic enterotoxin, cereolysin O, enolase) and others reduced (e.g., flagellin, L2 component of hemolysin BL, bacillolysin, sphingomyelinase, PC-PLC, PI-PLC, cytotoxin K) in the extracellular proteome of a ΔflhF mutant. Deprivation of FlhF also resulted in significant attenuation in the pathogenicity of this strain in an experimental model of infection in Galleria mellonella larvae. Our work highlights the multifunctional role of FlhF in B. cereus, being this protein involved in bacterial flagellation, swarming, protein secretion, and pathogenicity

Production, Secretion and Biological Activity of Bacillus cereus Enterotoxins

Bacillus cereus behaves as an opportunistic pathogen frequently causing gastrointestinal diseases, and it is increasingly recognized to be responsible for severe local or systemic infections. Pathogenicity of B. cereus mainly relies on the secretion of a wide array of toxins and enzymes and also on the ability to undergo swarming differentiation in response to surface-sensing. In this report, the pathogenicity exerted by B. cereus toxins is described with particular attention to the regulatory mechanisms of production and secretion of HBL, Nhe and CytK enterotoxins

NO RECOGNITION OF MHC CLASS II+ CELLS INFECTED WITH A VACCINIA VIRUS ENCODING INFLUENZA TYPE-A NUCLEOPROTEIN BY CLASS II-RESTRICTED T-CELLS

MHC class II is mostly charged by antigens derived from the outside of the antigen-presenting cell (APC), while class I presents endogenous antigens transported as peptides to the endoplasmic reticulum (ER) by specific transporters. Nevertheless, many antigens, especially glycoproteins, can be presented in vitro by class II even if endogenous. In order to investigate the class-II-restricted T-cell response to endogenously synthesized influenza nucleoprotein (NP) synthesized in infected cells as a model of non-glycosylated nuclear protein, class-II-restricted cytolytic T-cell (CTL) clones were established from BALB/c (H-2d) mice immunized with either influenza A/PR/8/34 (PR8) strain or with a vaccinia virus encoding the NP protein. Two of the clones were characterized in detail and turned out to be cytolytic, I-A(d)-restricted and NP peptide 218-229 specific. Even though an in vivo class-II-restricted T-cell response was elicited in BALB/c mice immunized with a vaccinia virus encoding nucleoprotein (Vacc-NP), class II+ mouse lymphoma cells were not lysed by the class-II-restricted clones in vitro when they were infected with the same virus or with a vaccinia virus encoding a truncated form of NP with no karyophilic sequence, showing that the de novo synthesized protein targeted to the nucleus or remaining in the cytoplasm cannot charge class II through the same pathway as class I in murine APC. These results extend previous observations made on transfected cells to cells that express an antigen during viral infection

Features of Bacillus cereus swarm cells

When propagated on solid surfaces, Bacillus cereus can produce differentiated swarm cells under a wide range of growth conditions. This behavioural versatility is ecologically relevant, since it allows this bacterium to adapt swarming to environmental changes. Swarming by B. cereus is medically important: swarm cells are more virulent and particularly prone to invade host tissues. Characterisation of swarming-deficient mutants highlights that flagellar genes as well as genes governing different metabolic pathways are involved in swarm-cell differentiation. In this review, the environmental and genetic requirements for swarming and the role played by swarm cells in the virulence this pathogen exerts will be outlined