[Sporadic lymphangioleiomyomatosis: A rare cause of bilateral spontaneous pneumothorax in young woman]. / La lymphangioléiomyomatose sporadique : cause rare du pneumothorax spontané bilatéral chez la femme jeune.

Sporadic lymphangioleiomyomatosis is an orphan disease of the young woman. Its clinical manifestations are mainly respiratory, including spontaneous pneumothorax. The management is multidisciplinary. We report the case of a young patient with bilateral spontaneous pneumothorax revealing sporadic lymphangioleiomyomatosis.

Modulation of host cell membrane fluidity: a novel mechanism for preventing bacterial adhesion.

Adhesion of bacterial enteropathogens to host mucosal surfaces is a critical primary step in the pathogenesis of diarrheal disease. We investigated the effects of altering the physical properties of eukaryotic cells on bacterial adhesion with the use of a series of three structurally dissimilar membrane fluidizers and several Escherichia coli as test strains. Lipid fluidity of the cell plasma membrane was measured by steady-state fluorescence anisotropy employing the probe 1-(4-trimethylammoniumphenyl)-6-phenyl-1,3, 5-hexatriene. There was a dose-dependent and reversible inhibition of bacterial adhesion with increasing membrane fluidity. Time course experiments indicated that increasing membrane fluidity during the early stages of bacterial adhesion was essential for inhibition of attachment. None of the fluidizers affected the viability of either eukaryotic or prokaryotic cells. These findings demonstrate, for the first time, that changes in plasma membrane physical properties of epithelial cells can prevent microbial adhesion. This also suggests that altering the membrane properties of host cells could form a basis for novel strategies to prevent bacterial adhesion during infection in vivo.

Divergent signal transduction responses to infection with attaching and effacing Escherichia coli.

Shiga toxin-producing Escherichia coli (STEC) O157:H7 is an attaching and effacing pathogen that causes hemorrhagic colitis and the hemolytic-uremic syndrome. Although this organism causes adhesion pedestals, the cellular signals responsible for the formation of these lesions have not been clearly defined. We have shown previously that STEC O157:H7 does not induce detectable tyrosine phosphorylation of host cell proteins upon binding to eukaryotic cells and is not internalized into nonphagocytic epithelial cells. In the present study, tyrosine-phosphorylated proteins were detected under adherent STEC O157:H7 when coincubated with the non-intimately adhering, intimin-deficient, enteropathogenic E. coli (EPEC) strain CVD206. The ability to be internalized into epithelial cells was also conferred on STEC O157:H7 when coincubated with CVD206 ([158 +/- 21] % of control). Neither the ability to rearrange phosphotyrosine proteins nor that to be internalized into epithelial cells was evident following coincubation with another STEC O157:H7 strain or with the nonsignaling espB mutant of EPEC. E. coli JM101(pMH34/pSSS1C), which overproduces surface-localized O157 intimin, also rearranged tyrosine-phosphorylated and cytoskeletal proteins when coincubated with CVD206. In contrast, JM101 (pMH34/pSSS1C) demonstrated rearrangement of cytoskeletal proteins, but not tyrosine-phosphorylated proteins, when coincubated with intimin-deficient STEC (strains CL8KO1 and CL15). These findings indicate that STEC O157:H7 forms adhesion pedestals by mechanisms that are distinct from those in attaching and effacing EPEC. Taken together, these findings point to diverging signal transduction responses to infection with attaching and effacing bacterial enteropathogens.

Heterogeneity in phenotypic and genotypic characteristics among strains of Hafnia alvei.

Hafnia alvei is an emerging human pathogen associated with sporadic cases and outbreaks of diarrhea. Bangladeshi isolates of H. alvei possess the Escherichia coli attaching and effacing (eaeA) gene and demonstrate an attaching and effacing phenotype. In the present study we examined 11 Canadian H. alvei isolates and strain 19,982 from Bangladesh to determine if the formation of attaching and effacing lesions is a property shared among multiple isolates. Attaching and effacing lesions were detected by induction of tyrosine kinase protein phosphorylation and cytoskeletal rearrangements in infected tissue culture epithelial cells with immunofluorescence microscopy and by the examination of infected cells with transmission electron microscopy. The presence of the eaeA gene was examined by PCR and colony blot hybridization. Profiles of outer membrane protein extracts, chromosomal macrorestriction fragments, and plasmids were also examined. Accumulation of host phosphotyrosine proteins and rearrangement of the cytoskeletal protein alpha-actinin were both observed in HEp-2 cells infected with H. alvei 19,982. In contrast, none of the other 11 clinical H. alvei isolates demonstrated either of these responses, nor did they form attaching and effacing lesions under electron microscopy. Consistent with the absence of the attaching and effacing phenotype, these clinical isolates did not possess the eaeA gene. The outer membrane protein profiles of all the Canadian isolates were identical but differed from that of H. alvei 19,982. Pulsed-field gel electrophoresis and plasmid profile analyses of the clinical H. alvei isolates differed substantially from those of the Bangladeshi strain. These results indicate that there is heterogeneity among H. alvei strains with respect to signal transduction, attaching and effacing adhesion, outer membrane constituents, and genotype. Epidemiological studies on enteropathogenic H. alvei thus need to go beyond simple species designations and require specific identification of the virulent clones.

Alpha-actinin accumulation in epithelial cells infected with attaching and effacing gastrointestinal pathogens.

Fluorescent F-actin staining utilizing phalloidin, a highly toxic mushroom poison, is used as an indirect test to detect attaching and effacing (AE) bacteria. A study was done to determine if accumulation of alpha-actinin in infected tissue culture cells is a consistent feature and whether it corresponds with the AE response. Rearrangement of alpha-actinin was detected using immunofluorescence microscopy by incubation of infected cells with a murine monoclonal anti-alpha-actinin antibody. Foci of alpha-actinin-specific fluorescence corresponding to areas of bacterial adhesion were detected by transmission electron microscopy in HEp-2 and gastric KATO-III cells infected with only those bacterial strains that formed AE lesions. Therefore, this study shows that alpha-actinin accumulation is a consistent, specific manifestation of the AE phenotype and forms the basis for the development of a safe alternative test for detecting AE bacteria.

Signal transduction responses following adhesion of verocytotoxin-producing Escherichia coli.

Attaching and effacing adhesion to epithelial cells is a pathognomonic feature of infection by both enteropathogenic Escherichia coli (EPEC) and certain strains of verocytotoxin-producing E. coli (VTEC). EPEC adhesion to tissue culture epithelial cells results in activation of the phosphatidylinositol pathway, with elevated levels of inositol 1,4,5-triphosphate and cytosolic free calcium. In this report, we show that VTEC also activate this signal transduction pathway in infected epithelial cells. Specifically, increased levels of inositol 1,4,5-triphosphate and intracellular free calcium were observed in HEp-2 cells infected with VTEC of serotype O157:H7. VTEC of serotypes O157:H7 and O113:H21 also induced increases in intracellular calcium levels in the human intestinal crypt-like cell line T84, even with minimal or no attaching and effacing activity as monitored by transmission electron microscopy. In contrast to EPEC, VTEC failed to induce tyrosine phosphorylation of epithelial cell proteins in HEp-2 and T84 cells, as determined by indirect immunofluorescence microscopy. These findings suggest that signal transduction responses to VTEC, including elevated levels of inositol triphosphates and intracellular free calcium, are independent of formation of the attaching and effacing lesion. Our findings also show that VTEC pathogenesis may involve signal transduction pathways that are distinct from those induced by EPEC infection.

Increased adherence of Escherichia coli RDEC-1 to human tissue culture cells results in the activation of host signaling pathways.

Attaching and effacing (AE) adhesion is associated with the pathogenesis of enteropathogenic Escherichia coli (EPEC) and rabbit diarrheogenic E. coli (RDEC-1). Although RDEC-1 provides an animal model for investigating pathophysiology of EPEC infection, RDEC-1 does not adhere to human cell lines, thereby limiting in vitro investigation. Therefore, transformed RDEC-1 strains expressing adhesins derived from human diarrheogenic E. coli were studied. These adhesins promoted AE adhesion of RDEC-1 and led to the accumulation of alpha-actinin aggregates in the cytoplasm of infected cells. Furthermore, these strains induced host signal transduction pathways, resulting in tyrosine phosphorylation of host proteins and an intracellular elevation of calcium. These results demonstrate that RDEC-1 and EPEC stimulate similar signal transduction pathways in infected epithelial cells, thus lending additional support for the use of RDEC-1 as a model for the study of human EPEC infection.

Distinct binding properties of eaeA-negative verocytotoxin-producing Escherichia coli of serotype O113:H21.

Infection of humans with verotoxin-producing Escherichia coli (VTEC) O113:H21 is associated with clinical features comparable to those associated with infection with attaching and effacing VTEC strains including those of serotype O157:H7. We have shown previously that the adhesion phenotype of VTEC O157:H7 is influenced by the presence of a homolog of the chromosomal eaeA (for E. coli attaching and effacing) gene. In contrast, by colony blot hybridization, VTEC O113:H21 is negative for the eaeA gene. Therefore, the aim of this study was to define the adhesion phenotype of VTEC O113:H21 strain CL-15 to both cultured epithelial cells (HEp-2) and rabbit intestine in vivo. Under transmission electron microscopy, areas of microvillus effacement were observed in regions directly beneath the organism in CL-15-infected cells both in vitro and in vivo. However, F-actin adhesion pedestals on the host plasma membrane were absent. Failure of CL-15 to induce polymerization of actin was confirmed by using staining of F-actin with fluorescein-labeled phalloidin. Under indirect immunofluorescence microscopy, CL-15-infected HEp-2 cells also failed to demonstrate the recruitment of another cytoskeletal element, alpha-actinin, below foci of bacterial adhesion. In contrast, VTEC O157:H7 infection of HEp-2 cells was associated with increased alpha-actinin immunofluorescence. These findings suggest that bacterial factors distinct from those of EaeA are necessary for the adhesion phenotype of VTEC O113:H21.