RÉSUMÉ
Limited evidence describing how host genetic variants affect the composition of the microbiota is currently available. The aim of this study was to assess the associations between a set of candidate host genetic variants and microbial composition in both saliva and gut in the TwinsUK registry. A total of 1,746 participants were included in this study and provided stool samples. A subset of 1,018 participants also provided self-reported periodontal data, and 396 of those participants provided a saliva sample. Host DNA was extracted from whole-blood samples and processed for Infinium Global screening array, focusing on 37 selected single-nucleotide polymorphisms (SNPs) previously associated with periodontitis. The gut and salivary microbiota of participants were profiled using 16S ribosomal RNA amplicon sequencing. Associations between genotype on the selected SNPs and microbial outcomes, including α diversity, ß diversity, and amplicon sequence variants (ASVs), were investigated in a multivariate mixed model. Self-reported periodontal status was also compared with microbial outcomes. Downstream analyses in gut microbiota and salivary microbiota were carried out separately. IL10 rs6667202 and VDR 2228570 SNPs were associated with salivary α diversity, and SNPs in IL10, HSA21, UHRF2, and Fc-γR genes were associated with dissimilarity matrix generated from salivary ß diversity. The SNP that was associated with the greatest number of salivary ASVs was VDR 2228570 followed by IL10 rs6667202, and that of gut ASVs was NPY rs2521364. There were 77 salivary ASVs and 39 gut ASVs differentially abundant in self-reported periodontal disease versus periodontal health. The dissimilarity between saliva and gut microbiota within individuals appeared significantly greater in self-reported periodontal cases compared to periodontal health. IL10 and VDR gene variants may affect salivary microbiota composition. Periodontal status may drive variations in the salivary microbiota and possibly, to a lesser extent, in the gut microbiota.
Sujet(s)
Microbiome gastro-intestinal , Microbiote , Parodontite , Humains , Microbiome gastro-intestinal/génétique , Interleukine-10 , Microbiote/génétique , Génotype , ARN ribosomique 16S/génétique , ARN ribosomique 16S/analyse , Ubiquitin-protein ligases/génétiqueRÉSUMÉ
AIMS: In order to develop complementary health management strategies for marine mammals, we used culture-based and culture-independent approaches to identify gastrointestinal lactobacilli of the common bottlenose dolphin, Tursiops truncatus. METHODS AND RESULTS: We screened 307 bacterial isolates from oral and rectal swabs, milk and gastric fluid, collected from 38 dolphins in the U.S. Navy Marine Mammal Program, for potentially beneficial features. We focused our search on lactobacilli and evaluated their ability to modulate TNF secretion by host cells and inhibit growth of pathogens. We recovered Lactobacillus salivarius strains which secreted factors that stimulated TNF production by human monocytoid cells. These Lact. salivarius isolates inhibited growth of selected marine mammal and human bacterial pathogens. In addition, we identified a novel Lactobacillus species by culture and direct sequencing with 96·3% 16S rDNA sequence similarity to Lactobacillus ceti. CONCLUSIONS: Dolphin-derived Lact. salivarius isolates possess features making them candidate probiotics for clinical studies in marine mammals. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study to isolate lactobacilli from dolphins, including a novel Lactobacillus species and a new strain of Lact. salivarius, with potential for veterinary probiotic applications. The isolation and identification of novel Lactobacillus spp. and other indigenous microbes from bottlenose dolphins will enable the study of the biology of symbiotic members of the dolphin microbiota and facilitate the understanding of the microbiomes of these unique animals.
Sujet(s)
Grand dauphin/microbiologie , Lactobacillus/isolement et purification , Probiotiques/isolement et purification , Animaux , Lactobacillus/classification , Lactobacillus/génétique , ARN ribosomique 16S/génétique , Facteurs de nécrose tumorale/biosynthèseRÉSUMÉ
The recently discovered pathogen Bordetella holmesii has been isolated from the airways and blood of diseased humans. Genetic events contributing to the emergence of B. holmesii are not understood, and its phylogenetic position among the bordetellae remains unclear. To address these questions, B. holmesii strains were analyzed by comparative genomic hybridization (CGH) to a Bordetella pertussis microarray and by multilocus sequence typing. Both methods indicated substantial sequence divergence between B. pertussis and B. holmesii. However, CGH identified a putative pathogenicity island of 66 kb that is highly conserved between these species and contains several IS481 elements that may have been laterally transferred from B. pertussis to B. holmesii. This island contains, among other genes, a functional, iron-regulated locus encoding the biosynthesis, export, and uptake of the siderophore alcaligin. The acquisition of this genomic island by B. holmesii may have significantly contributed to its emergence as a human pathogen. Horizontal gene transfer between B. pertussis and B. holmesii may also explain the unusually high sequence identity of their 16S rRNA genes.
Sujet(s)
Bordetella pertussis/classification , Bordetella pertussis/génétique , Bordetella/classification , Bordetella/génétique , Ilots génomiques/génétique , Protéines bactériennes/génétique , Protéines bactériennes/métabolisme , Bordetella/isolement et purification , Bordetella pertussis/isolement et purification , Évolution moléculaire , Génome bactérien , Humains , Acides hydroxamiques/métabolisme , Données de séquences moléculaires , Hybridation d'acides nucléiques , Séquençage par oligonucléotides en batterie , Phylogenèse , Analyse de séquence d'ADNRÉSUMÉ
The Bordetella master virulence regulatory system, BvgAS, controls a spectrum of gene expression states, including the virulent Bvg(+) phase, the avirulent Bvg(-) phase, and at least one Bvg-intermediate (Bvg(i)) phase. We set out to define the species- and strain-specific features of this regulon based on global gene expression profiling. Rather than functioning as a switch, Bvg controls a remarkable continuum of gene expression states, with hundreds of genes maximally expressed in intermediate phases between the Bvg(+) and Bvg(-) poles. Comparative analysis of Bvg regulation in B. pertussis and B. bronchiseptica revealed a relatively conserved Bvg(+) phase transcriptional program and identified previously uncharacterized candidate virulence factors. In contrast, control of Bvg(-)- and Bvg(i)-phase genes diverged substantially between species; regulation of metabolic, transporter, and motility loci indicated an increased capacity in B. bronchiseptica, compared to B. pertussis, for ex vivo adaptation. Strain comparisons also demonstrated variation in gene expression patterns within species. Among the genes with the greatest variability in patterns of expression, predicted promoter sequences were nearly identical. Our data suggest that the complement of transcriptional regulators is largely responsible for transcriptional diversity. In support of this hypothesis, many putative transcriptional regulators that were Bvg regulated in B. bronchiseptica were deleted, inactivated, or unregulated by BvgAS in B. pertussis. We propose the concept of a "flexible regulon." This flexible regulon may prove to be important for pathogen evolution and the diversification of host range specificity.
Sujet(s)
Protéines bactériennes/génétique , Bordetella bronchiseptica/génétique , Régulation de l'expression des gènes bactériens , Régulon , Facteurs de transcription/génétique , Protéines bactériennes/métabolisme , Bordetella bronchiseptica/métabolisme , Transduction du signal , Spécificité d'espèce , Facteurs de transcription/métabolismeRÉSUMÉ
Pathogens of the bacterial genus Bordetella cause respiratory disease in humans and animals. Although virulence and host specificity vary across the genus, the genetic determinants of this diversity remain unidentified. To identify genes that may underlie key phenotypic differences between these species and clarify their evolutionary relationships, we performed a comparative analysis of genome content in 42 Bordetella strains by hybridization of genomic DNA to a microarray representing the genomes of three Bordetella species and by subtractive hybridization. Here we show that B. pertussis and B. parapertussis are predominantly differentiated from B. bronchiseptica by large, species-specific regions of difference, many of which encode or direct synthesis of surface structures, including lipopolysaccharide O antigen, which may be important determinants of host specificity. The species also exhibit sequence diversity at a number of surface protein-encoding loci, including the fimbrial major subunit gene, fim2. Gene loss, rather than gene acquisition, accompanied by the proliferation of transposons, has played a fundamental role in the evolution of the pathogenic bordetellae and may represent a conserved evolutionary mechanism among other groups of microbial pathogens.
Sujet(s)
Bordetella/classification , Bordetella/pathogénicité , Génomique , Animaux , Antigènes bactériens/génétique , Bordetella/génétique , Bordetelloses/microbiologie , Évolution moléculaire , Protéines de fimbriae/génétique , Humains , Données de séquences moléculaires , Hybridation d'acides nucléiques/méthodes , Séquençage par oligonucléotides en batterie , Phylogenèse , Analyse de séquence d'ADN , Spécificité d'espèce , Facteurs de virulence des Bordetella/génétiqueRÉSUMÉ
Cyclospora cayetanensis is a coccidian parasite which causes severe gastroenteritis in humans. Molecular information on this newly emerging pathogen is scarce. Our objectives were to assess genetic variation within and between human-associated C. cayetanensis and baboon-associated Cyclospora papionis by examining the internal transcribed spacer (ITS) region of the ribosomal RNA operon, and to develop an efficient polymerase chain reaction- (PCR)-based method to distinguish C. cayetanensis from other closely related organisms. For these purposes, we studied C. cayetanensis ITS-1 nucleotide variability in 24 human faecal samples from five geographic locations and C. papionis ITS-1 variability in four baboon faecal samples from Tanzania. In addition, a continuous sequence encompassing ITS-1, 5.8S rDNA and ITS-2 was determined from two C. cayetanensis samples. The results indicate that C. cayetanensis and C. papionis have distinct ITS-1 sequences, but identical 5.8S rDNA sequences. ITS-1 is highly variable within and between samples, but variability does not correlate with geographic origin of the samples. Despite this variability, conserved species-specific ITS-1 sequences were identified and a single-round, C. cayetanensis-specific PCR-based assay with a sensitivity of one to ten oocysts was developed. This consistent and remarkable diversity among Cyclospora spp. ITS-1 sequences argues for polyparasitism and simultaneous transmission of multiple strains.
Sujet(s)
Cyclospora/génétique , ADN des protozoaires/génétique , Espaceur de l'ADN ribosomique/génétique , Animaux , Séquence nucléotidique , Cyclospora/composition chimique , Cyclospora/classification , Cyclosporose/parasitologie , ADN des protozoaires/composition chimique , ADN des protozoaires/isolement et purification , ADN ribosomique/composition chimique , ADN ribosomique/génétique , Espaceur de l'ADN ribosomique/composition chimique , Variation génétique , Humains , Données de séquences moléculaires , Phylogenèse , Réaction de polymérisation en chaîne/méthodes , ARN ribosomique 5.8S/génétique , Similitude de séquences d'acides nucléiques , Spécificité d'espèce , Opéron d'ARNr/génétiqueRÉSUMÉ
The characterization of life is immeasurably enhanced by determination of complete genome sequences. For organisms that engage in intimate interactions with others, the genome sequence from one participant, and associated tools, provide unique insight into its partner. We discuss how the human genome sequence will further our understanding of microbial pathogens and commensals, and vice versa. We also propose criteria for implicating a host gene in microbial pathogenesis, and urge consideration of a'second human genome project'.
Sujet(s)
Bactéries/génétique , Génome bactérien , Génome humain , Bactéries/pathogénicité , Humains , Analyse de séquence , VirulenceRÉSUMÉ
Bordetella pertussis, the agent of whooping cough, is capable of invading human respiratory epithelial cells. In this study, we investigated the mechanisms by which B. pertussis invades the human lung epithelial cell line A549 and normal human bronchial epithelial (NHBE) cells. In vitro adhesion and invasion assays using both cell types with a virulent B. pertussis strain and its isogenic mutants revealed profound defects in a mutant deficient in filamentous hemagglutinin (FHA) expression. In addition, a mutant in which an FHA Arg-Gly-Asp (RGD) site had been changed to Arg-Ala-Asp had significantly diminished invasiveness, although its adhesiveness was comparable to that of the parental strain. Furthermore, a synthetic RGD-containing hexapeptide inhibited invasion of both cell types by the virulent strain. These results demonstrate that an RGD sequence of FHA is involved in B. pertussis invasion of epithelial cells in vitro. Monoclonal antibodies directed against human alpha5beta1 integrin, but not other integrins, blocked invasion, indicating that this integrin is involved in B. pertussis invasion. Taken together, these findings suggest that B. pertussis FHA may promote invasion of human respiratory epithelial cells through the interaction of its RGD sequence with host cell alpha5beta1 integrin.
Sujet(s)
Adhésines bactériennes/immunologie , Antigènes bactériens/immunologie , Bordetella pertussis/pathogénicité , Cellules épithéliales/microbiologie , Hémagglutinines/immunologie , Récepteur fibronectine/immunologie , Muqueuse respiratoire/microbiologie , Facteurs de virulence des Bordetella , Adhésines bactériennes/génétique , Anticorps monoclonaux/pharmacologie , Antigènes bactériens/génétique , Adhérence bactérienne/effets des médicaments et des substances chimiques , Bordetella pertussis/immunologie , Hémagglutinines/génétique , Humains , Mutation , Oligopeptides/synthèse chimique , Oligopeptides/pharmacologie , Muqueuse respiratoire/cytologie , Cellules cancéreuses en culture , VirulenceRÉSUMÉ
Real-time PCR methods with primers and a probe targeting conserved regions of the bacterial 16S ribosomal DNA (rDNA) revealed a larger amount of rDNA in blood specimens from healthy individuals than in matched reagent controls. However, the origins and identities of these blood-associated bacterial rDNA sequences remain obscure.
Sujet(s)
ADN bactérien/sang , ADN ribosomique/sang , Réaction de polymérisation en chaîne/méthodes , ARN ribosomique 16S/génétique , Bactéries/génétique , Amorces ADN/génétique , HumainsRÉSUMÉ
Filamentous hemagglutinin (FHA) is a dominant cell surface-associated Bordetella pertussis adhesin. Recognition that this protein is secreted in significant amounts and that bacterial adhesins may have other activities, prompted an assessment of FHA effects on human macrophages. Incubation of human macrophage-like U937 cells with preparations of FHA resulted in dose-dependent cytotoxicity, with death of 95% of treated cells after 24 h. Based on the use of four independent methods, death of these cells could be largely attributed to apoptosis. FHA-associated apoptosis was also observed in THP-1 macrophage-like cells, fresh human peripheral blood monocyte-derived macrophages (MDM), and BEAS-2B human bronchial epithelial cells. Infection of MDM with wild-type B. pertussis resulted in apoptosis within 6 h, while infection with an FHA-deficient derivative strain was only 50% as effective. FHA-associated cytotoxicity was preceded by host cell secretion of tumor necrosis factor alpha (TNF-alpha), a potential proapoptotic factor. However, pretreatment of cells with a neutralizing anti-TNF-alpha monoclonal antibody inhibited only 16% of the FHA-associated apoptosis. On the other hand, a blocking monoclonal antibody directed against TNF-alpha receptor 1 inhibited FHA-associated apoptosis by 47.7% (P = 0.0001), suggesting that this receptor may play a role in the death pathway activated by FHA. Our in vitro data indicate that secreted and cell-associated FHA elicits proinflammatory and proapoptotic responses in human monocyte-like cells, MDM, and bronchial epithelial cells and suggest a previously unrecognized role for this prominent virulence factor in the B. pertussis-host interaction.
Sujet(s)
Adhésines bactériennes/toxicité , Apoptose/effets des médicaments et des substances chimiques , Bordetella pertussis/pathogénicité , Hémagglutinines/toxicité , Inflammation/étiologie , Facteurs de virulence des Bordetella , Adhérence bactérienne , Lignée cellulaire , Relation dose-effet des médicaments , Humains , Lipopolysaccharides/toxicité , Facteur de nécrose tumorale alpha/métabolisme , VirulenceRÉSUMÉ
Whipple's disease is caused by a cultivation-resistant bacterium, Tropheryma whippelii. Ultrastructural studies of intestinal biopsy specimens from patients with Whipple's disease have shown that intracellular and extracellular bacteria are present, but the preferred site of growth is unknown. Tissue sections from 8 patients with Whipple's disease and from 19 healthy control subjects were analyzed by use of fluorescence in situ hybridization and laser scanning confocal microscopy, to determine the location of rRNA that would indicate the presence of metabolically active bacteria. T. whippelii rRNA was most prevalent near the tips of intestinal villi, in the lamina propria, just basal to epithelial cells. Most of the bacterial rRNA signal appeared to be located between cells and did not colocalize with the human intracellular protein vimentin. The location of bacterial rRNA in tissues from patients with Whipple's disease provides evidence that bacteria are growing outside cells and suggests that T. whippelii is not an obligate intracellular pathogen.
Sujet(s)
Actinobacteria/isolement et purification , Muqueuse intestinale/microbiologie , ARN bactérien/analyse , ARN ribosomique/analyse , Maladie de Whipple/microbiologie , Actinobacteria/génétique , Actinobacteria/croissance et développement , Biopsie , Humains , Hybridation fluorescente in situ , Muqueuse intestinale/anatomopathologie , Muqueuse intestinale/ultrastructure , Microvillosités/microbiologie , Microvillosités/anatomopathologie , Microvillosités/ultrastructure , Valeurs de référence , Vimentine/analyse , Maladie de Whipple/anatomopathologieRÉSUMÉ
BACKGROUND: Little is known about the pathogenesis of Whipple disease, the reservoirs of Tropheryma whippelii, and the proportion of persons harboring the bacterium without "classic" intestinal abnormalities. OBJECTIVE: To assess the presence of T. whippelii in patients undergoing upper endoscopy for a variety of indications. DESIGN: Prospective and routine diagnostic examination of patients. SETTING: Three academic medical centers in California; Minnesota; and Heidelberg, Germany. PATIENTS: 342 patients undergoing endoscopy for evaluation of dyspepsia or possible peptic ulcer (group A, 173 patients), malabsorption (group B, 37 patients), or clinical suspicion of Whipple disease (group C, 132 patients). MEASUREMENTS: Small-intestinal biopsy specimens were tested by polymerase chain reaction for T. whippelii DNA and examined for histopathologic abnormalities. RESULTS: All patients with negative histologic findings also had negative results for T. whippelii DNA. CONCLUSIONS: T. whippelii occurs only rarely in intestinal mucosa that lacks histopathologic evidence of Whipple disease. The human small intestinal mucosa is an unlikely reservoir for this organism.
Sujet(s)
Infections à Actinomycetales/microbiologie , Actinomycetales/isolement et purification , ADN bactérien/analyse , Muqueuse intestinale/microbiologie , Maladie de Whipple/microbiologie , Infections à Actinomycetales/diagnostic , Réservoirs de maladies , Dyspepsie/anatomopathologie , Endoscopie , Humains , Muqueuse intestinale/anatomopathologie , Intestin grêle/microbiologie , Intestin grêle/anatomopathologie , Syndromes de malabsorption/anatomopathologie , Ulcère peptique/anatomopathologie , Réaction de polymérisation en chaîne , Études prospectives , Maladie de Whipple/diagnosticRÉSUMÉ
Functional genomic technologies such as high density DNA microarrays allow biologists to study the structure and behavior of thousands of genes in a single experiment. One of the fields in which microarrays have had an increasingly important impact is host-pathogen interactions. Early investigations in this area over the past two years not only emphasize the utility of this approach, but also highlight the stereotyped gene expression responses of different host cells to diverse infectious stimuli, and the potential value of broad dataset comparisons in revealing fundamental features of innate immunity. The comparative analysis of recently published datasets involving human gene expression responses to two bacterial respiratory pathogens illustrates many of these points. Comparisons between these large, highly parallel sets of experimental observations also emphasize important technical and experimental design issues as future challenges.
Sujet(s)
Bordetella pertussis/génétique , Génome bactérien , Séquençage par oligonucléotides en batterie , Pseudomonas aeruginosa/génétique , Bordetella pertussis/pathogénicité , ADN bactérien/analyse , Humains , Pseudomonas aeruginosa/pathogénicitéRÉSUMÉ
Bordetella pertussis, the causative agent of whooping cough, has many well-studied virulence factors and a characteristic clinical presentation. Despite this information, it is not clear how B. pertussis interaction with host cells leads to disease. In this study, we examined the interaction of B. pertussis with a human bronchial epithelial cell line (BEAS-2B) and measured host transcriptional profiles by using high-density DNA microarrays. The early transcriptional response to this pathogen is dominated by altered expression of cytokines, DNA-binding proteins, and NFkappaB-regulated genes. This previously unrecognized response to B. pertussis was modified in similar but nonidentical fashions by the antiinflammatory agents dexamethasone and sodium salicylate. Cytokine protein expression was confirmed, as was neutrophil chemoattraction. We show that B. pertussis induces mucin gene transcription by BEAS-2B cells then counters this defense by using mucin as a binding substrate. A set of genes is described for which the catalytic activity of pertussis toxin is both necessary and sufficient to regulate transcription. Host genomic transcriptional profiling, in combination with functional assays to evaluate subsequent biological events, provides insight into the complex interaction of host and pathogen.
Sujet(s)
Bordetella pertussis/physiologie , Appareil respiratoire/microbiologie , Transcription génétique , Bordetella pertussis/pathogénicité , Lignée cellulaire , Chimiokines/biosynthèse , Cellules épithéliales/métabolisme , Cellules épithéliales/microbiologie , Humains , Mucines/métabolisme , Facteur de transcription NF-kappa B/métabolisme , Séquençage par oligonucléotides en batterie , Toxine pertussique , Poly(ADP-ribose) polymerases/métabolisme , Appareil respiratoire/métabolisme , Facteurs de virulence des Bordetella/métabolisme , Coqueluche/anatomopathologieRÉSUMÉ
Complete genomic sequences of microbial pathogens and hosts offer sophisticated new strategies for studying host-pathogen interactions. DNA microarrays exploit primary sequence data to measure transcript levels and detect sequence polymorphisms, for every gene, simultaneously. The design and construction of a DNA microarray for any given microbial genome are straightforward. By monitoring microbial gene expression, one can predict the functions of uncharacterized genes, probe the physiologic adaptations made under various environmental conditions, identify virulence-associated genes, and test the effects of drugs. Similarly, by using host gene microarrays, one can explore host response at the level of gene expression and provide a molecular description of the events that follow infection. Host profiling might also identify gene expression signatures unique for each pathogen, thus providing a novel tool for diagnosis, prognosis, and clinical management of infectious disease.
Sujet(s)
Bactéries , Régulation de l'expression des gènes bactériens/génétique , Génétique microbienne/tendances , Génomique/méthodes , Algorithmes , Animaux , Bactéries/génétique , Bactéries/pathogénicité , Génomique/tendances , Génotype , HumainsRÉSUMÉ
Whipple's disease is a systemic disorder associated with a cultivation-resistant, poorly characterized actinomycete, Tropheryma whippelii. We determined a nearly complete rRNA operon sequence of T. whippelii from specimens from 3 patients with Whipple's disease, as well as partial operon sequences from 43 patients. Variability was observed in the 16S-23S rRNA spacer sequences, leading to the description of five distinct sequence types. One specimen contained two spacer sequence types, raising the possibility of a double infection. Secondary structure models for the primary rRNA transcript and mature rRNAs revealed rare or unique features.
Sujet(s)
Actinomycetales/génétique , Opéron , ARN ribosomique/génétique , Maladie de Whipple/microbiologie , Actinomycetales/classification , Séquence nucléotidique , Variation génétique , Humains , Données de séquences moléculaires , Conformation d'acide nucléique , Phylogenèse , ARN ribosomique 16S/génétique , ARN ribosomique 23S/génétique , ARN ribosomique 5S/génétique , Spécificité d'espèceRÉSUMÉ
Rhinosporidium seeberi, a microorganism that can infect the mucosal surfaces of humans and animals, has been classified as a fungus on the basis of morphologic and histochemical characteristics. Using consensus polymerase chain reaction (PCR), we amplified a portion of the R. seeberi 18S rRNA gene directly from infected tissue. Analysis of the aligned sequence and inference of phylogenetic relationships showed that R. seeberi is a protist from a novel clade of parasites that infect fish and amphibians. Fluorescence in situ hybridization and R. seeberi- specific PCR showed that this unique 18S rRNA sequence is also present in other tissues infected with R. seeberi. Our data support the R. seeberi phylogeny recently suggested by another group. R. seeberi is not a classic fungus, but rather the first known human pathogen from the DRIPs clade, a novel clade of aquatic protistan parasites (Ichthyosporea).
Sujet(s)
Eucaryotes/classification , Gènes d'ARN ribosomique , ARN ribosomique 18S/génétique , Rhinosporidiose/microbiologie , Rhinosporidium/classification , Animaux , Clonage moléculaire , Maladies des chiens/microbiologie , Chiens , Eucaryotes/génétique , Eucaryotes/isolement et purification , Humains , Hybridation fluorescente in situ , Microscopie électronique , Données de séquences moléculaires , Phylogenèse , Réaction de polymérisation en chaîne , Rhinosporidiose/médecine vétérinaire , Rhinosporidium/génétique , Rhinosporidium/isolement et purification , Rhinosporidium/ultrastructure , Analyse de séquence d'ADNRÉSUMÉ
Innate immune responses to pathogens are believed to be patterned and stereotyped. Adaptive responses display variety but in relatively few types of products and with limited numbers of mechanisms. Is this apparent disparity between microbial pathogen diversity and a restricted set of host responses an accurate picture of infection or is it the result of a limited collection of analytic tools? DNA microarray technology permits one to address simple descriptive questions about global gene expression inside cells. In particular, it offers an opportunity to examine the relationship between host and pathogen in much greater detail than has been possible previously. One can now ask, firstly, how a host cell or organism 'sees' a microbial pathogen from the viewpoint of gene expression responses and, secondly, at what level it is able to discriminate between different agents. Other potential insights to be reaped include the identification of microbial determinants of the host response, the temporal features of the 'conversation' between host and pathogen, novel strategies for therapeutic and prophylactic intervention and prognostic markers of outcome.
Sujet(s)
Analyse de profil d'expression de gènes , Régulation de l'expression des gènes , Système immunitaire/immunologie , Infections/immunologie , Séquençage par oligonucléotides en batterie , Animaux , Infections bactériennes/immunologie , Traitement automatique des données , Humains , Système immunitaire/métabolisme , Activation des lymphocytes , Souris , Cellules U937/immunologie , Virulence , Maladies virales/immunologieRÉSUMÉ
Molecular, sequence-based environmental surveys of microorganisms have revealed a large degree of previously uncharacterized diversity. However, nearly all studies of the human endogenous bacterial flora have relied on cultivation and biochemical characterization of the resident organisms. We used molecular methods to characterize the breadth of bacterial diversity within the human subgingival crevice by comparing 264 small subunit rDNA sequences from 21 clone libraries created with products amplified directly from subgingival plaque, with sequences obtained from bacteria that were cultivated from the same specimen, as well as with sequences available in public databases. The majority (52.5%) of the directly amplified 16S rRNA sequences were <99% identical to sequences within public databases. In contrast, only 21.4% of the sequences recovered from cultivated bacteria showed this degree of variability. The 16S rDNA sequences recovered by direct amplification were also more deeply divergent; 13.5% of the amplified sequences were more than 5% nonidentical to any known sequence, a level of dissimilarity that is often found between members of different genera. None of the cultivated sequences exhibited this degree of sequence dissimilarity. Finally, direct amplification of 16S rDNA yielded a more diverse view of the subgingival bacterial flora than did cultivation. Our data suggest that a significant proportion of the resident human bacterial flora remain poorly characterized, even within this well studied and familiar microbial environment.