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PubMed2006

Helicobacter pylori phase variation, immune modulation and gastric autoimmunity.

Helicobacter pylori can be regarded as a model pathogen for studying persistent colonization of humans. Phase-variable expression of Lewis blood-group antigens by H. pylori allows this microorganism to modulate the host T-helper-1-cell versus T-helper-2-cell response. We describe a model in which interactions between host lectins and pathogen carbohydrates facilitate asymptomatic persistence of H. pylori. This delicate balance, favourable for both the pathogen and the host, could lead to gastric autoimmunity in genetically susceptible individuals.

Amino Acid Sequence↗
PubMed1993

Environmental regulation of the fim switch controlling type 1 fimbrial phase variation in Escherichia coli K-12: effects of temperature and media.

Expression of type 1 fimbriae in Escherichia coli K-12 is phase variable and associated with the inversion of a short DNA element (switch). The fim switch requires either fimB (on-to-off or off-to-on switching) or fimE (on-to-off switching only) and is affected by the global regulators leucine-responsive regulatory protein (Lrp), integration host factor (IHF), and H-NS. Here it is shown that switching frequencies are regulated by both temperature and media and that these effects appear to be independent. fimE-promoted on-to-off switching occurs far more rapidly than previously estimated (0.3 per cell per generation in defined rich medium at 37 degrees C) and faster at lower than at higher temperatures. In direct contrast, fimB-promoted switching increases with temperature, with optima between 37 and 41 degrees C. Switching promoted by both fimB and fimE is stimulated by aliphatic amino acids (alanine, isoleucine, leucine, and valine), and this stimulation requires lrp. Furthermore, lrp appears to differentially regulate fimB- and fimE-promoted switching in different media.

Agar↗
PubMed2005

Phase variation mediated niche adaptation during prolonged experimental murine infection with Helicobacter pylori.

Changes in the repeats associated with the recently redefined repertoire of 31 phase-variable genes in Helicobacter pylori were investigated following murine gastric colonization for up to one year in three unrelated H. pylori strains. Between the beginning and end of the experimental period, changes were seen in ten genes (32 %), which would alter gene expression in one or more of the three strains studied. For those genes that showed repeat length changes at the longest time points, intermediate time points showed differences between the rates of change for different functional groups of genes. Genes most likely to be associated with immediate niche fitting changed most rapidly, including phospholipase A (pldA) and LPS biosynthetic genes. Other surface proteins, which may be under adaptive immune selection, changed more slowly. Restriction-modification genes showed no particular temporal pattern. The number of genes that phase varied during adaptation to the murine gastric environment correlated inversely with their relative fitness as previously determined in this murine model of colonization. This suggests a role for these genes in determining initial fitness for colonization as well as in subsequent niche adaptation. In addition, a coding tandem repeat within a phase-variable gene which does not control actual gene expression was also investigated. This repeat was found to vary in copy number during colonization. This suggests that changes in the structures encoded by tandem repeats may also play a role in altered protein functions and/or immune evasion during H. pylori colonization.

Adaptation, Physiological↗
PubMed1996

Tetrameric repeat units associated with virulence factor phase variation in Haemophilus also occur in Neisseria spp. and Moraxella catarrhalis.

The tetrameric repeat units 5'-CAAT-3' and 5'-GCAA-3' are associated with phase variable expression of lipopolysaccharide biosynthetic genes in Haemophilus influenzae. Four other tetrameric repeat units have also been reported from H. influenzae strain Rd, 5'-CAAC-3', 5'-GACA-3', 5'-AGCT-3', and 5'-TTTA-3', which are also associated with putative virulence factors. Using oligonucleotide probes corresponding to five tandem copies of each of these tetramers, we have screened three strains of Neisseria meningitidis and one each of Neisseria gonorrhoeae, Neisseria lactamica, Haemophilus parainfluenzae, Bordetella pertussis, Bordetella parapertussis, Bordetella bronchiceptica and Moraxella catarrhalis for the presence of these motifs. We have demonstrated the presence of multiple copies of the 5'-GCAA-3' motif in all the Neisseria strains tested, and also the repeated motif 5'-CAAC-3' in M. catarrhalis. We have further demonstrated by Southern blot analysis that the 5'-CAAC-3' repeats detected in M. catarrhalis are probably associated with the same genes as in H. influenzae, but that the 5'-GCAA-3' motifs in N. meningitidis are not. The use of characterised tetrameric DNA sequences as hybridisation probes may prove useful in the identification of novel phase variable virulence determinants in organisms other than H. influenzae.

Base Sequence↗
PubMed1983

Phase variation of hyaluronidase-producing peptostreptococci associated with periodontal disease.

Peptostreptococci isolated from diseased periodontal pockets produced large amounts of extracellular hyaluronidase throughout their growth cycle. Phase variants, obtained by re-streaking aged colonies of the wild types, had different colonial morphology and shorter generation times. These variants produced less hyaluronidase, and enzyme production occurred only during the early stationary phase.

Humans↗
PubMed1987

Phase variation in nasal airways resistance assessed by active anterior rhinomanometry.

Measurements of nasal airways resistance in normal subjects have shown a significant difference between values obtained in the two respiratory phases. Higher overall values are found during expiration in both the unprepared and decongested nose but these differences fail to maintain statistical significance after application of a decongestant. This indicates that the phase of respiration in which measurement is made must be routinely recorded especially when measurement is made without decongestant, and comparisons of data made only with recordings from equivalent parts of the nasal cycle.

Adolescent↗
PubMed1990

Disulfide-linked protein associated with Mycoplasma pneumoniae cytadherence phase variation.

Wild-type Mycoplasma pneumoniae possessed a protein with a very high molecular weight under nonreducing conditions (greater than 340,000; designated HMW5); this protein was absent from a noncytadhering phase variant lacking HMW1, 2, 3, and 4. When examined by two-dimensional nonreducing-reducing gel electrophoresis, HMW5 dissociated to yield a single polypeptide spot of molecular weight 190,000 that comigrated with cytadherence phase-variable protein HMW2. Extraction of wild-type mycoplasmas with Triton X-100 revealed the exclusive partitioning of HMW5 with the detergent-insoluble cytoskeletonlike triton shell.

Antigens, Bacterial↗
PubMed1977

Ultrastructural investigations on surface structures involved in Coxiella burnetii phase variation.

By using the cytochemical staining procedure with concanavalin A, horseradish peroxidase, and diaminobenzidine, no surface carbohydrates with terminal alpha-glucosyl or sterically closely related residues could be detected on the cell walls of Coxiella burnetii phases I and II. Using a polycationized ferritin derivative as a cytochemical probe, anionic binding sites were visualized in the electron microscope on cell membranes of C. burnetii phase II, but not on phase I organisms. The sites appeared to be masked in phase I particles. Anionic sites could be demonstrated on phase I organisms after treatment with NaIO4 or dimethyl sulfoxide. A number of different biological properties of C. burnetii phases I and II may depend on the presence or absence of a net negative charge on the surface of the cell walls of these organisms.

Acetone↗
PubMed1994

Growth phase variation of integration host factor level in Escherichia coli.

We have measured the intracellular abundance of integration host factor (IHF), a site-specific, heterodimeric DNA-binding protein, in exponential- and stationary-phase cultures of Escherichia coli K-12. Western immunoblot analysis showed that cultures that had been growing exponentially for several generations contained 0.5 to 1.0 ng of IHF subunits per microgram of total protein and that this increased to 5 to 6 ng/microgram in late-stationary-phase cultures. IHF is about one-third to one-half as abundant in exponentially growing cells as HU, a structurally related protein that binds DNA with little or no site specificity. Wild-type IHF is metabolically stable, but deletion mutations that eliminated one subunit reduced the abundance of the other when cells enter stationary phase. We attribute this reduction to the loss of stabilizing interactions between subunits. A mutation that inactivates IHF function but not subunit interaction increased IHF abundance, consistent with results of previous work showing that IHF synthesis is negatively autoregulated. We estimate that steady-state exponential-phase cultures contain about 8,500 to 17,000 IHF dimers per cell, a surprisingly large number for a site-specific DNA-binding protein with a limited number of specific sites. Nevertheless, small reductions in IHF abundance had significant effects on several IHF-dependent functions, suggesting that the wild-type exponential phase level is not in large excess of the minimum required for occupancy of physiologically important IHF-binding sites.

Amino Acid Sequence↗
PubMed2000

Evaluation of phase variation of nontypeable Haemophilus influenzae lipooligosaccharide during nasopharyngeal colonization and development of otitis media in the chinchilla model.

Nontypeable Haemophilus influenzae (NTHI) has four loci, lic-1 to lic-3 and lgtC, that generate phase-variable lipooligosaccharide (LOS) structures. lic-1, which is required for the expression of phosphorylcholine (ChoP), is the best characterized and is associated with an enhanced ability of H. influenzae to persist within the nasopharynges of infant rats. Recent data indicate that LOS impacts various aspects of NTHI virulence in the chinchilla model of nasopharyngeal colonization and otitis media (OM). In this study the effects of ChoP expression and the sequences of lic-1 to lic-3 and lgtC of NTHI strain 2019 were evaluated in the chinchilla OM model. Nasopharyngeal colonization data showed that a switch from the ChoP(-) to the ChoP(+) phenotype was observed as early as day 3 after intranasal inoculation. Chinchillas colonized by strains with the ChoP(+) phenotype demonstrated a significantly higher level of NTHI 2019 per milliliter of nasal lavage fluid than chinchillas colonized with predominantly the ChoP(-) variant (P < 0.05). The concentration of cells with the ChoP(+) phenotype in the middle ear was 3 log units higher than that of cells with the ChoP(-) variant (P < 0.01). There was a statistically significant association between ChoP(+) expression in the nasal lavage and the development of OM with culture-positive middle ear fluids in this model. These data suggest that expression of the ChoP(+) phenotype promotes enhanced nasopharyngeal colonization and development of OM.

Animals↗
PubMed1985

Luteal phase variations in endogenous concentrations of prostaglandins PGE and PGF and in the capacity for their in vitro formation in the human corpus luteum.

One evidence for a luteolytic role for prostaglandin F2 alpha in the human is the increase in luteal PGF at times corresponding to luteolysis as reported earlier by us and other groups. There have been other contradictory reports on this point. In the present experiments we have measured the concentrations of PGE and PGF in 16 more human corpora lutea and have determined the capacity of those tissues to form PGE and PGF in vitro. PGF concentrations were highest in the mid luteal phase but were accompanied by high PGE concentrations. On the other hand, in the late luteal phase PGF concentrations, lower than in mid luteal but generally higher than in early luteal phase, were significantly higher than PGE concentrations. This pattern in PGE and PGF concentrations was also evident in the capacity of these tissues to form these compounds in vitro. In view of the known capacity of PGE2 to counteract the luteolytic effect of PGF2 alpha, these variations in the relative concentrations of PGE and PGF during the luteal phase may be of significance in the process of luteolysis in the human.

Corpus Luteum↗
PubMed1996

Estrous cycle phase variations in visceromotor and cardiovascular responses to colonic distension in the anesthetized rat.

Visceromotor and cardiovascular responses to colonic distension were measured in female rats, anesthetized with halothane in oxygen, in the proestrus, estrus, metestrus and diestrus phases of the estrous cycle. Ten rats were studied in each group and responses were measured at 5-min intervals for 60 min. A mixed model analysis of variance showed that there was no real change in either the visceral or cardiovascular response with time. There was a highly significant difference in visceromotor responses between the phases of the estrous cycle (P < 0.001). During the phase of proestrus the balloon pressure at which a response was triggered was much lower, with a mean value (95% confidence interval) of 18.7 (16.1, 21.8) mmHg, than the other phases with mean values (95% confidence interval) of 31.9 (27.4, 37.2) mmHg for estrus, 28.1 (24.2, 32.8) mmHg for metestrus, and 31.1 (26.7, 36.3) mmHg for diestrus. The mean arterial blood pressure increased in all groups (range 3.2, 5.4%) as a response to the stimulus, but there was no associated heart rate variability and no significant differences in cardiovascular changes between the groups (P = 0.6). The visceromotor responses measured during the phase of proestrus occurred at a significantly lower threshold than in the other phases of estrous.

Anesthesia↗
PubMed2002

Quenching of n,pi*-excited states in the gas phase: variations in absolute reactivity and selectivity.

The quenching of the n,pi*-excited azoalkane 2,3-diazabicyclo[2.2.2]oct-2-ene by 19 heteroatom-containing electron and hydrogen donors, that is, amines, sulfides, ethers, and alcohols, was investigated in the gas phase. Deuterium isotope effects were measured for 9 selectively deuterated derivatives. The data support the involvement of an excited charge-transfer complex, that is, an exciplex, for tertiary amines and sulfides, and a competitive direct hydrogen transfer from the C-H bonds of ethers or from the N-H or O-H bonds of secondary and primary amines or alcohols. The recently observed "inverted" solvent effect for the fluorescence quenching of azoalkanes by amines and sulfides in solution is supported by the observed rate constants in the gas phase, which are substantially larger than those in solution. A more pronounced inverted solvent effect for the weaker electron-donating sulfides and a presumably faster exciplex deactivation result in a switch-over in absolute reactivity relative to tertiary amines in the gas phase. Most importantly, the kinetic data demonstrate that the reactivity of the strongly dipolar O-H and N-H bonds in photoinduced hydrogen abstraction reactions shows a larger decrease upon solvation than that of the less polar C-H bonds. The azoalkane data are compared with previous studies on quenching of n,pi*-triplet-excited ketones in the gas phase.

Journal Article↗
PubMed1980

Phase variation in the genus Serratia.

During an investigation of the serotypes of Serratia marcescens present in a cancer centre, it was found that 8.7% of 241 strains had more than one H antigen. There were 19 diphasic and two triphasic strains. Antigen H1 was a component of the phases in the approximately half of the strains with multiple phases. This is the first report of such phases in the genus Serratia.

Antigens, Bacterial↗
PubMed1994

Lipopolysaccharide smooth-rough phase variation in bacteria of the genus Chlamydia.

In two strains of Chlamydia psittaci and in Chlamydia trachomatis serotype L1, we have detected a so-far-unknown antigen which (i) is resistant to heat and proteolytic digestion, (ii) can be extracted with phenol-water into the water phase, (iii) gives a ladder-like banding pattern in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, (iv) is immunogenic in rabbits and mice, and (v) contains immunoreactivity of lipid A, a common and characteristic component of gram-negative lipopolysaccharides (LPS). Thus, chlamydiae contain, in addition to the known rough-type LPS, another LPS type which is phenotypically smooth (S-LPS). S-LPS was observed preferentially in chlamydiae grown in the yolk sac of embryonated eggs; it was, however, also detected by immunofluorescence in tissue culture-grown chlamydiae with a monoclonal antibody against S-LPS.

Animals↗
PubMed1990

Phenotypic switching in mycoplasmas: phase variation of diverse surface lipoproteins.

The ability of some microorganisms to rapidly alter the expression and structure of surface components reflects an important strategy for adaptation to changing environments, including those encountered by infectious agents within respective host organisms. Mycoplasma hyorhinis, a wall-less prokaryotic pathogen of the class Mollicutes, is shown to undergo high-frequency phase transitions in colony morphology and opacity and in the expression of diverse lipid-modified, cell-surface protein antigens. These proteins spontaneously vary in size, contain highly repetitive structures, and are oriented with their carboxyl-terminal region external to the membrane. Thus, mycoplasma membrane lipoproteins generate microbial surface diversity and may be part of a complex system that controls interactions of these organisms with their hosts.

Antibodies, Monoclonal↗
PubMed1998

High-frequency intracellular infection and erythrogenic toxin A expression undergo phase variation in M1 group A streptococci.

A clonal variant of serotype M1 group A streptococcus, strain 90-131, disseminated to several continents, where it was associated with severe systemic infections and toxic shock. Although this strain harbours the speA gene and is efficiently internalized by human epithelial cells, clinical isolates often fail to express the erythrogenic toxin under laboratory growth conditions. Cultures of strain 90-131 were observed to phase vary between small, dry, compact and larger, more mucoid colonies. The former were shown to be poorly internalized by epithelial cells. Analysis of RNA by Northern hybridization demonstrated that the emml, hasA and speA genes were weakly transcribed in cultures derived from the small colonies and highly transcribed in those derived from the large colonies. An insertion mutation in mga (the multigene activator) downregulated the invasion of epithelial cells and the transcription of emm1 and hasA, but had little impact on the transcription of speA. These are the first data to suggest the existence of a common regulatory circuit linking intracellular invasion, M protein, hyaluronic acid capsule and erythrogenic toxin expression by group A streptococcus. Moreover, the genetic instability of toxin expression exhibited by this serotype may impact on laboratory studies that attempt to associate toxin production with toxic shock.

Antigens, Bacterial↗