RESUMO
OBJECTIVE: Helicobacter pylori (Hp) is a major risk factor for gastric cancer (GC). Hp promotes DNA damage and proteasomal degradation of p53, the guardian of genome stability. Hp reduces the expression of the transcription factor USF1 shown to stabilise p53 in response to genotoxic stress. We investigated whether Hp-mediated USF1 deregulation impacts p53-response and consequently genetic instability. We also explored in vivo the role of USF1 in gastric carcinogenesis. DESIGN: Human gastric epithelial cell lines were infected with Hp7.13, exposed or not to a DNA-damaging agent camptothecin (CPT), to mimic a genetic instability context. We quantified the expression of USF1, p53 and their target genes, we determined their subcellular localisation by immunofluorescence and examined USF1/p53 interaction. Usf1-/- and INS-GAS mice were used to strengthen the findings in vivo and patient data examined for clinical relevance. RESULTS: In vivo we revealed the dominant role of USF1 in protecting gastric cells against Hp-induced carcinogenesis and its impact on p53 levels. In vitro, Hp delocalises USF1 into foci close to cell membranes. Hp prevents USF1/p53 nuclear built up and relocates these complexes in the cytoplasm, thereby impairing their transcriptional function. Hp also inhibits CPT-induced USF1/p53 nuclear complexes, exacerbating CPT-dependent DNA damaging effects. CONCLUSION: Our data reveal that the depletion of USF1 and its de-localisation in the vicinity of cell membranes are essential events associated to the genotoxic activity of Hp infection, thus promoting gastric carcinogenesis. These findings are also of clinical relevance, supporting USF1 expression as a potential marker of GC susceptibility.
Assuntos
Carcinogênese , Mucosa Gástrica , Infecções por Helicobacter/metabolismo , Helicobacter pylori , Neoplasias Gástricas , Proteína Supressora de Tumor p53/genética , Fatores Estimuladores Upstream/metabolismo , Animais , Carcinogênese/genética , Carcinogênese/metabolismo , Linhagem Celular , Dano ao DNA , Mucosa Gástrica/metabolismo , Mucosa Gástrica/microbiologia , Mucosa Gástrica/patologia , Instabilidade Genômica , Helicobacter pylori/metabolismo , Helicobacter pylori/patogenicidade , Humanos , Camundongos , Complexo de Endopeptidases do Proteassoma/metabolismo , Neoplasias Gástricas/genética , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/microbiologia , UbiquitinaçãoRESUMO
Cfr is a radical S-adenosyl-l-methionine (SAM) enzyme that confers cross-resistance to antibiotics targeting the 23S rRNA through hypermethylation of nucleotide A2503. Three cfr-like genes implicated in antibiotic resistance have been described, two of which, cfr(B) and cfr(C), have been sporadically detected in Clostridium difficile However, the methylase activity of Cfr(C) has not been confirmed. We found cfr(B), cfr(C), and a cfr-like gene that shows only 51 to 58% protein sequence identity to Cfr and Cfr-like enzymes in clinical C. difficile isolates recovered across nearly a decade in Mexico, Honduras, Costa Rica, and Chile. This new resistance gene was termed cfr(E). In agreement with the anticipated function of the cfr-like genes detected, all isolates exhibited high MIC values for several ribosome-targeting antibiotics. In addition, in vitro assays confirmed that Cfr(C) and Cfr(E) methylate Escherichia coli and, to a lesser extent, C. difficile 23S rRNA fragments at the expected positions. The analyzed isolates do not have mutations in 23S rRNA genes or genes encoding the ribosomal proteins L3 and L4 and lack poxtA, optrA, and pleuromutilin resistance genes. Moreover, these cfr-like genes were found in Tn6218-like transposons or integrative and conjugative elements (ICE) that could facilitate their transfer. These results indicate selection of potentially mobile cfr-like genes in C. difficile from Latin America and provide the first assessment of the methylation activity of Cfr(C) and Cfr(E), which belong to a cluster of Cfr-like proteins that does not include the functionally characterized enzymes Cfr, Cfr(B), and Cfr(D).
Assuntos
Clostridioides difficile/genética , Genes Bacterianos , Proteínas de Bactérias/genética , Clostridioides difficile/efeitos dos fármacos , Clostridioides difficile/isolamento & purificação , Infecções por Clostridium/tratamento farmacológico , Infecções por Clostridium/epidemiologia , Infecções por Clostridium/microbiologia , Farmacorresistência Bacteriana Múltipla/genética , Humanos , Sequências Repetitivas Dispersas , América Latina/epidemiologia , Testes de Sensibilidade Microbiana , Epidemiologia Molecular , Filogenia , RNA Bacteriano/genética , RNA Ribossômico 23S/genéticaRESUMO
BACKGROUND & AIMS: Helicobacter pylori infection is the main risk factor for gastric cancer. We characterized the interactions of H pylori with gastric epithelial progenitor and stem cells in humans and mice and investigated how these interactions contribute to H pylori-induced pathology. METHODS: We used quantitative confocal microscopy and 3-dimensional reconstruction of entire gastric glands to determine the localizations of H pylori in stomach tissues from humans and infected mice. Using lineage tracing to mark cells derived from leucine-rich repeat-containing G-protein coupled receptor 5-positive (Lgr5(+)) stem cells (Lgr5-eGFP-IRES-CreERT2/Rosa26-TdTomato mice) and in situ hybridization, we analyzed gastric stem cell responses to infection. Isogenic H pylori mutants were used to determine the role of specific virulence factors in stem cell activation and pathology. RESULTS: H pylori grow as distinct bacterial microcolonies deep in the stomach glands and interact directly with gastric progenitor and stem cells in tissues from mice and humans. These gland-associated bacteria activate stem cells, increasing the number of stem cells, accelerating Lgr5(+) stem cell proliferation, and up-regulating expression of stem cell-related genes. Mutant bacteria with defects in chemotaxis that are able to colonize the stomach surface but not the antral glands in mice do not activate stem cells. In addition, bacteria that are unable to inject the contact-dependent virulence factor CagA into the epithelium colonized stomach glands in mice, but did not activate stem cells or produce hyperplasia to the same extent as wild-type H pylori. CONCLUSIONS: H pylori colonize and manipulate the progenitor and stem cell compartments, which alters turnover kinetics and glandular hyperplasia. Bacterial ability to alter the stem cells has important implications for gastrointestinal stem cell biology and H pylori-induced gastric pathology.
Assuntos
Mucosa Gástrica/microbiologia , Infecções por Helicobacter/microbiologia , Helicobacter pylori/crescimento & desenvolvimento , Receptores Acoplados a Proteínas G/metabolismo , Células-Tronco/microbiologia , Animais , Antígenos de Bactérias/genética , Antígenos de Bactérias/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Biomarcadores/metabolismo , Proliferação de Células , Modelos Animais de Doenças , Mucosa Gástrica/metabolismo , Genótipo , Infecções por Helicobacter/imunologia , Infecções por Helicobacter/patologia , Helicobacter pylori/genética , Helicobacter pylori/patogenicidade , Interações Hospedeiro-Patógeno , Humanos , Hiperplasia , Cinética , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Mutação , Organoides , Fenótipo , Receptores Acoplados a Proteínas G/genética , Células-Tronco/metabolismo , Células-Tronco/patologia , Técnicas de Cultura de Tecidos , VirulênciaRESUMO
The majority of the world population carry the gastric pathogen Helicobacter pylori. Fortunately, most individuals experience only low-grade or no symptoms, but in many cases the chronic inflammatory infection develops into severe gastric disease, including duodenal ulcer disease and gastric cancer. Here we report on a protective mechanism where H. pylori attachment and accompanying chronic mucosal inflammation can be reduced by antibodies that are present in a vast majority of H. pylori carriers. These antibodies block binding of the H. pylori attachment protein BabA by mimicking BabA's binding to the ABO blood group glycans in the gastric mucosa. However, many individuals demonstrate low titers of BabA blocking antibodies, which is associated with an increased risk for duodenal ulceration, suggesting a role for these antibodies in preventing gastric disease.
RESUMO
OBJECTIVE: To study the association between anti-VacA antibodies and pre-neoplastic lesions (IM), gastric cancer (GC), and duodenal ulcer (DU). METHODS: A case-control study that included 347 patients, 90 with IM, 60 with GC, 52 with DU, and 145 with non-atrophic gastritis was conducted. For the analysis, a polytomous logistic regression models were used. Anti-VacA antibodies were identified in sera from these patients, either by Western blot assay (WB), using antigens produced by H. pylori s1m1 strain, or by neutralization assay challenging HeLa cells with H. pylori VacA s1m1 cytotoxin. RESULTS: Results of the WB assay showed no association between WB-anti-VacA antibodies and gastroduodenal diseases. In contrast, when antibodies that neutralize VacA cytotoxic activity were studied, a significant association was found with IM (OR 2.7, 95% CI 1.4-5.1) and DU (OR 2.3, 95% CI 1.1-4.9) and an even stronger association with GC (OR 3.9, 95% CI 1.8-8.5). A significant association with histological subtypes of GC (diffuse and intestinal) and of IM (complete and incomplete) was also found. In addition, the association showed a significant dose-response effect in the case of GC, but not of DU or IM. These associations did not change substantially after adjustment for confounding factors. MAIN CONCLUSION: This study showed that VacA-neutralizing antibodies are significantly associated with gastroduodenal diseases, especially GC, and that they might be used as risk markers of GC and DU.
Assuntos
Anticorpos Antibacterianos/sangue , Anticorpos Neutralizantes/sangue , Proteínas de Bactérias/imunologia , Úlcera Duodenal/imunologia , Úlcera Duodenal/microbiologia , Neoplasias Gástricas/imunologia , Neoplasias Gástricas/microbiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Antígenos de Bactérias/imunologia , Estudos de Casos e Controles , Feminino , Infecções por Helicobacter/imunologia , Helicobacter pylori/imunologia , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Razão de Chances , Fatores de RiscoRESUMO
BACKGROUND: There is an association of interleukin (IL)1B polymorphism with gastric cancer risk. However systematic reviews of the existing evidence have shown that such association varies across populations with different genetic ancestry. AIM: To evaluate the association of IL-1B-511 and IL-1RN polymorphism and Helicobacter pylori IgG antibodies CagA, with gastric cancer in two Colombian cities located in a high risk area for gastric cancer. MATERIAL AND METHODS: A case-control study including 46 gastric cancer cases and 99 controls with non-atrophic gastritis from a high risk zone for gastric cancer. Polymorphism genotyping was carried out by polymerase chain reaction (PCR) and IgG CagA status by ELISA. RESULTS: IgG CagA seropositive individuals had an increased gastric cancer risk (odds ratio (OR) = 11.56; 95% confidence intervals (CI) 2.62-50.91 in Tunja and OR = 19.66, 95%CI 0.98-395 in Bogotá). IL-1B-511TT carriers in Tunja had increased risk of gastric cancer (OR = 11.31; 95%CI 1.20-106.54)), while IL-1RN*2 alelle carriers in Bogotá showed an inverse association with gastric cancer risk (OR = 0.03; 95%CI 0.01-0.65). CONCLUSIONS: This study adds evidence to the positive association of Helicobacter pylori CagA positive strains with non-cardial gastric cancer etiology. There is a possible heterogeneity in the association of IL-1B gene polymorphism with cancer, in populations of similar ethnic background and settled in the same risk area.
Assuntos
Antígenos de Bactérias/genética , Proteínas de Bactérias/genética , Infecções por Helicobacter/genética , Helicobacter pylori/imunologia , Proteína Antagonista do Receptor de Interleucina 1/genética , Interleucina-1beta/genética , Polimorfismo Genético/genética , Neoplasias Gástricas , Adulto , Idoso , Estudos de Casos e Controles , Colômbia/etnologia , Feminino , Infecções por Helicobacter/imunologia , Helicobacter pylori/genética , Humanos , Imunoglobulina G/sangue , Masculino , Pessoa de Meia-Idade , Fatores de Risco , Neoplasias Gástricas/genética , Neoplasias Gástricas/imunologia , Neoplasias Gástricas/microbiologiaRESUMO
Hungatella hathewayi has been observed to be a member of the gut microbiome. Unfortunately, little is known about this organism in spite of being associated with human fatalities; it is important to understand virulence mechanisms and epidemiological prospective to cause disease. In this study, a patient with chronic neurologic symptoms presented to the clinic with subsequent isolation of a strain with phenotypic characteristics suggestive of Clostridium difficile. However, whole-genome sequence found the organism to be H. hathewayi. Analysis including publicly available Hungatella genomes found substantial genomic differences as compared to the type strain, indicating this isolate was not C. difficile. We examined the whole-genome of Hungatella species and related genera, using comparative genomics to fully examine species identification and toxin production. Orthogonal phylogenetic using the 16S rRNA gene and entire genome analyses that included genome distance analyses using Genome-to-Genome Distance (GGDC), Average Nucleotide Identity (ANI), and a pan-genome analysis with inclusion of available public genomes determined the speciation to be Hungatella. Two clearly differentiated groups were identified, one including a reference H. hathewayi genome (strain DSM-13,479) and a second group that was determined to be H. effluvii, which included our clinical isolate. Also, some genomes reported as H. hathewayi were found to belong to other genera, including Clostridium and Faecalicatena. We show that the Hungatella species have an open pan-genome reflecting high genomic diversity. This study highlights the importance of correctly assigning taxonomic identification, particularly in disease-associated strains, to better understand virulence and therapeutic options.
Assuntos
Clostridiaceae , Genoma Bacteriano , Filogenia , Clostridiaceae/genética , RNA Ribossômico 16S/genéticaRESUMO
Tumour Necrosis Factor (TNF) and Heat Shock Protein 70 (HSP70) are important molecules in inflammatory, infectious and tumoral processes. The genes codifying these molecules are polymorphic and certain alleles have been associated with susceptibility to disease. Gastric cancer is associated with an Helicobacter pylori-induced chronic inflammatory response. The aim of this work was to analyze whether polymorphisms in inflammation-related genes are associated with the development of gastric cancer. We studied 447 Mexican adult patients including 228 with non-atrophic gastritis, 98 with intestinal metaplasia, 63 with gastric cancer and 58 with duodenal ulcer, and 132 asymptomatic individuals as well. DNA from peripheral white blood cells was typed for the Single Nucleotide Polymorphisms (SNPs) -308 of TNF-alpha, +252 of TNF-beta, +190 of HSP70-1, +1267 of HSP70-2 and +2437 of HSP70-HOM. Compared with the asymptomatic group, we found a significant association of TNF-beta*A and HSP70-1*C alleles with gastric cancer (OR 5.69 and 3.76, respectively) and HSP70-1*C with duodenal ulcer (OR 3.08). Genotype TNF-beta G/G showed a significant gene-dose effect with gastric cancer (OR 0.09); whereas HSP70-1 C/G showed significant association with both, gastric cancer (OR 13.31) and duodenal ulcer (OR 16.19). Polymorphisms in TNF and HSP70 showed a significant severity-dose-response as risk markers from preneoplastic lesions to gastric cancer in Mexican population, probably because of their association with an intense and sustained inflammatory response.
Assuntos
Úlcera Duodenal/genética , Predisposição Genética para Doença , Proteínas de Choque Térmico HSP70/genética , Lesões Pré-Cancerosas/genética , Neoplasias Gástricas/genética , Fator de Necrose Tumoral alfa/genética , Adulto , Úlcera Duodenal/microbiologia , Úlcera Duodenal/patologia , Feminino , Genótipo , Haplótipos , Infecções por Helicobacter/complicações , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Polimorfismo de Nucleotídeo Único , Lesões Pré-Cancerosas/microbiologia , Lesões Pré-Cancerosas/patologia , Fatores de Risco , Neoplasias Gástricas/microbiologia , Neoplasias Gástricas/patologiaRESUMO
Clostridioides difficile is a Gram positive, sporulated, rod-shape, anaerobic pathogen responsible for nosocomial diarrhea and colitis, mainly in antibiotic treated patients. C. difficile produce two toxins responsible for disease, toxin A (TcdA) and toxin B (TcdB), although not all strains produce them. Non-toxigenic C. difficile (NTCD) strains are able to colonize the intestinal mucosa and are often isolated from asymptomatic individuals. NTCD are poorly studied, their evolutionary history has not been elucidated, and their relationship with illness remains controversial. The aim of this work was to analyze the phenotype of NTCD strains isolated from clinical cases in hospitals of México, and whether NTCD strains present characteristics that differentiate them from the toxigenic strains. Seventy-four C. difficile strains isolated from patients were tested for cytotoxicity and 14 were identified as NTCD strains. We analyzed phenotypical characteristics that are important for the biology of C. difficile like colony morphology, antibiotic resistance, motility, sporulation, and adherence. Strains were also genotyped to determine the presence of genes coding for TcdA, TcdB and binary toxin and ribotyped for 027 type. When compared with toxigenic strains, NTCD strains presented an enlarged branched colony morphology, higher resistance to metronidazole, and increased sporulation efficiency. This phenotype has been reported associated with mutations that regulates phenotypic characteristics like swimming, sporulation or adhesion. Our results show that phenotype of NTCD strains is heterogeneous but still present characteristics that differentiate them from toxigenic strains.
RESUMO
Helicobacter pylori (HP) genetics may determine its clinical outcomes. Despite high prevalence of HP infection in Latin America (LA), there have been no phylogenetic studies in the region. We aimed to understand the structure of HP populations in LA mestizo individuals, where gastric cancer incidence remains high. The genome of 107 HP strains from Mexico, Nicaragua and Colombia were analyzed with 59 publicly available worldwide genomes. To study bacterial relationship on whole genome level we propose a virtual hybridization technique using thousands of high-entropy 13 bp DNA probes to generate fingerprints. Phylogenetic virtual genome fingerprint (VGF) was compared with Multi Locus Sequence Analysis (MLST) and with phylogenetic analyses of cagPAI virulence island sequences. With MLST some Nicaraguan and Mexican strains clustered close to Africa isolates, whereas European isolates were spread without clustering and intermingled with LA isolates. VGF analysis resulted in increased resolution of populations, separating European from LA strains. Furthermore, clusters with exclusively Colombian, Mexican, or Nicaraguan strains were observed, where the Colombian cluster separated from Europe, Asia, and Africa, while Nicaraguan and Mexican clades grouped close to Africa. In addition, a mixed large LA cluster including Mexican, Colombian, Nicaraguan, Peruvian, and Salvadorian strains was observed; all LA clusters separated from the Amerind clade. With cagPAI sequence analyses LA clades clearly separated from Europe, Asia and Amerind, and Colombian strains formed a single cluster. A NeighborNet analyses suggested frequent and recent recombination events particularly among LA strains. Results suggests that in the new world, H. pylori has evolved to fit mestizo LA populations, already 500 years after the Spanish colonization. This co-adaption may account for regional variability in gastric cancer risk.
Assuntos
Evolução Molecular , Genoma Bacteriano/genética , Helicobacter pylori/genética , Helicobacter pylori/patogenicidade , Filogenia , África , América , Ásia , Proteínas de Bactérias/genética , Análise por Conglomerados , Impressões Digitais de DNA , Sondas de DNA , Bases de Dados de Ácidos Nucleicos , Europa (Continente) , Variação Genética , Genótipo , Humanos , Tipagem de Sequências Multilocus , Filogeografia , Homologia de Sequência do Ácido Nucleico , Virulência/genéticaRESUMO
Helicobacter pylori is one of the most common bacterial infections worldwide. It is associated with chronic gastritis, peptic ulcer disease and constitutes a major risk factor for gastric adenocarcinoma and lymphoma. The aim of this study was to evaluate the specific serologic immunoglobulin G (IgG) response to whole cells proteins, CagA and urease antigens of Helicobacter pylori in a Venezuelan population. We evaluated 66 patients from the Hospital Universitario de Caracas, attending in the gastroscopy service. H. pylori infection was detected by culture and rapid urease test. IgG antibodies against, CagA and ureases were tested by enzyme-linked immunosorbent assay method using highly purified recombinant antigens. We demonstrated the presence of H. pylori in 48/66 (72.7%), by culture and rapid urease test. We found a seroprevalence of 45 (68%) to whole cells, 34/66 (51%) to CagA and 18/66 (27%) to urease. The positive rates of CagA antibodies in patients with gastric ulcer, gastric cancer and chronic gastritis were 87.8%, 77.7% y 40.8% respectively. The serum antibodies anti-CagA were similar between peptic ulcer disease and gastric cancer patients.
Assuntos
Antígenos de Bactérias/imunologia , Proteínas de Bactérias/imunologia , Imunoglobulina G/imunologia , Gastropatias/imunologia , Gastropatias/microbiologia , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Urease/imunologia , VenezuelaRESUMO
Biomarkers may facilitate detection of gastric cancer at an earlier stage and reduce mortality. Here we sought to determine if the glycosylation profile of serum immunoglobulin G (IgG) could distinguish patients with non-atrophic gastritis (NAG), duodenal ulcer (DU) and gastric cancer (GC). Serum IgG was released and analyzed using nano-LC-TOF mass spectrometry. Statistically significant false discovery rate (FDR)-adjusted p-values were observed for 18 glycans, eight that differed significantly between NAG and GC, three that distinguished NAG from DU, and eight that differed between DU and GC. The IgG glycosylation signature may be useful as a predictive marker for gastric cancer.
Assuntos
Infecções por Helicobacter/complicações , Helicobacter pylori/patogenicidade , Neoplasias Gástricas/etiologia , Adolescente , Adulto , Distribuição por Idade , Idoso , Antígenos de Bactérias/imunologia , Proteínas de Bactérias/imunologia , Criança , Feminino , Infecções por Helicobacter/epidemiologia , Humanos , Masculino , México/epidemiologia , Pessoa de Meia-Idade , Sistema de Registros , Estudos Soroepidemiológicos , Distribuição por Sexo , Neoplasias Gástricas/mortalidadeRESUMO
Objetivo: Caracterizar los niveles de pepsinógeno y evaluar la capacidad de discriminación del PGI y la relación PGI/PGII para el diagnóstico serológico de atrofia gástrica en diferentes poblaciones colombianas. Materiales y métodos: Participaron 600 sujetos sin sintomatología gástrica y se analizaron 544 muestras de pacientes con sintomatología gástrica provenientes de diferentes poblaciones con riesgos opuestos para cáncer gástrico. A todos los participantes se les tomó muestra de sangre. En los pacientes se obtuvieron biopsias de antro y cuerpo para su diagnóstico inicial de lesiones gastroduodenales. Los niveles de pepsinógeno y la serología de Helicobacter pylori se estimaron con pruebas de ELISA. Los análisis estadísticos incluyeron pruebas de Kruskal-Wallis y Mann-Whitney, curva ROC y valores diagnósticos. Resultados: Los niveles de pepsinógeno en pacientes y sujetos asintomáticos difieren según la zona de riesgo de procedencia. Los niveles de PGI, PGII y PGI/PGII disminuyeron a medida que aumenta la severidad del diagnóstico histológico (p < 0,005), al igual que con el grado de severidad de la atrofia y la localización multifocal (p≤0,001). El PGI ≤86,68 y PGI/PGII ≤3,19 con un área bajo la curva de 0,76 identificó pacientes con atrofia severa multifocal, serología positiva para H. pylori y procedentes de la zona de riesgo alto, con sensibilidad de 77,5% y especificidad de 71,74%. Conclusión: Los resultados sugieren que los niveles de PGI, PGI/PGII conjuntamente con serología H. pylori positiva podrían ser considerados para la detección de atrofia severa en pacientes de la zona de riesgo alto. Se necesita otros estudios en poblaciones de riesgo alto.
Objective: To characterize levels of pepsinogen and evaluate the discrimination ability of pepsinogen I (PGI) and the PGI/ pepsinogen II (PGII) ratio for the serological diagnosis of gastric atrophy in different Colombian populations. Methods: A total 600 subjects without gastric symptoms participated and 544 samples from patients with gastric symptomatology were analyzed from different populations with opposing risks to gastric cancer. A blood sample was taken from all participants; a gastric antrum and body biopsy for the initial diagnosis of gastroduodenal lesions was obtained from the patients. The levels of pepsinogen and Helicobacter pylori serology were estimated with ELISA. Statistical analyses included Kruskal -Wallis and Mann -Whitney test, ROC curve and diagnostic values. Results: The levels of PGI and PGI / PGII differ by risk area of origin. Levels of PGI, PGII and PGI / PGII decreased with increasing severity of histological diagnosis (P < .005), as with the severity of atrophy and multifocal localization (P ≤.001). The PGI ≤ 86.68 and PGI / PGII ≤ 3.19 with an area under the curve of 0.76 identified patients with severe multifocal atrophy, positive serology for H. pylori, and from the high risk area, with a sensitivity of 77.5% and specificity of 71.74%. Conclusion: The results suggest that PGI levels together with PGI / PGII ratios and positive serology for H. pylori could be considered for the detection of severe atrophy in high-risk areas. Further studies are needed in high-risk populations © 2014 Instituto Nacional de Cancerología. Published by Elsevier España, S.L.U. All rights reserved.
Assuntos
Humanos , Neoplasias Gástricas , Testes Sorológicos , Helicobacter pylori , Pepsinogênio A , Pepsinogênio C , Atrofia , Ensaio de Imunoadsorção Enzimática , Sensibilidade e Especificidade , DiagnósticoRESUMO
Objetivo: Evaluar la asociación de los polimorfismos de alguna de las citocinas más estudiadas en relación con el cáncer gástrico (IL-1B-511, IL-1RN intron-2-VNTR, TNF-α-308, IL-10-819 e IL-10- 1082) y la presencia de anticuerpos hacia la proteína cagA de Helicobacter pylori con las lesiones preneoplásicas gástricas en pacientes colombianos. Materiales y métodos: Se estudiaron 185 pacientes con lesiones preneoplásicas (gastritis atrófca, metaplasia intestinal y displasia), y 154 controles (gastritis no atrófica), provenientes de hospitales de una zona de riesgo alto y otra de riesgo bajo para cáncer gástrico. Se obtuvieron biopsias gástricas y muestras de sangre; la genotipificación de los polimorfismos se hizo por discriminación alélica usando PCR en tiempo real y por PCR convencional y electroforesis en agarosa (VNTR del intron 2 de IL-1RN); la serología de Helicobacter pylori y Helicobacter pylori cagA se determinó por ELISA. Se utilizó regresión logística multinomial en el análisis estadístico. Resultados: El genotipo IL-1B-511TT (odds ratio = 4,05; intervalo de confianza 95% 1,35-12,10) se asoció a metaplasia intestinal; no se observaron otras asociaciones entre los diferentes polimorfismos y las lesiones preneoplásicas. La infección por Helicobacter pylori cagA positivo se asoció a gastritis atrófica, metaplasia intestinal y displasia (OR = 2,66; 13,70; 40,29, respectivamente). Conclusión: Los resultados sugieren que entre los genotipos proinflamatorios el genotipo IL-1B-511TT estaría asociado a la metaplasia intestinal, y la serología de Helicobacter pylori cagA positivo sería un biomarcador útil para intervenir y prevenir la presencia de lesiones preneoplásicas. Se necesitan otros estudios con población colombiana que evalúen la asociación hallada de IL1B-511 con la metaplasia intestinal.
Objective: To evaluate the relationship of some of the most studied cytokines (IL-1B-511, IL-1RN intron-2-VNTR, TNF-a-308, IL-10-819, and IL-10-1082) with gastric cancer, as well as the presence of anti-Helicobacter pylori cagA IgG antibodies with pre-cancerous lesions in Colombian patients. Materials and methods: A study was conducted on 185 patients with pre-cancerous lesions (atrophic gastritis, intestinal metaplasia and dysplasia), and 154 controls (non-atrophic gastritis), seen in hospitals in a high risk area, and another in a low risk area, for gastric cancer. Gastric biopsy specimens and blood samples were obtained. The genotyping of the polymorphisms was performed by allelic discrimination using real-time PCR, conventional PCR, and agarose electrophoresis (VNTR of IL-1RN intron 2). The serology of Helicobacter pylori and Helicobacter pylori cagA was determined by ELISA. A multinomial logistic regression was used in the statistical analysis. Results: The IL-1B-511TT genotype was associated with intestinal metaplasia (OR=4.05; 95% CI; 1.35-12.10). No other relationships were observed between the different polymorphisms and preimg/revistas/rcc/cancerous lesions. Infection due to a positive Helicobacter pylori cagA was associated with atrophic gastritis, intestinal metaplasia and dysplasia (OR=2.66; 13.70; 40.29, respectively). Conclusion: The results suggest that, among the pro-inflammatory genotypes, the IL-1B-511TT would be associated with intestinal metaplasia, and that a positive Helicobacter pylori cagA serology could be a useful biomarker for the intervention and prevention of pre-cancerous lesions. Further studies are required in the Colombian population in order to evaluate the relationship found between IL1B-511 and intestinal metaplasia.
Assuntos
Humanos , Lesões Pré-Cancerosas , Neoplasias Gástricas , Helicobacter pylori , Interleucina-10 , Sangue , Imunoglobulina G , Reação em Cadeia da Polimerase , Reação em Cadeia da Polimerase em Tempo RealRESUMO
Objetivo: Determinar la asociación entre los polimorfismos IL-1B-511, IL-1RN, TNF-α-308, IL-10-819 e IL-101082 y la infección por Helicobacter pylori CagA positivo en un grupo de pacientes con cáncer gástrico y úlcera duodenal en diferentes poblaciones en Colombia. Métodos: Estudio de casos y controles con 341 pacientes: con gastritis no atrófica, 194; con cáncer gástrico, 58; úlcera duodenal con lesiones preneoplásicas, 54; y con úlcera duodenal, 35. La genotipificación de los polimorfismos se hizo por discriminación alélica usando PCR en tiempo real, y la del IL-1RN, por PCR convencional y electroforesis en agarosa. La infección por Helicobacter pylori CagA se determinó mediante ELISA. Se utilizó la regresión logística en el análisis estadístico. Resultados: Ser portador del genotipo IL-1B-511TT (OR=4,69; IC 95% 1,22-18,09) y tener una infección por Helicobacter pylori CagA positivo (OR=4,43; IC 95% 1,72-11,4) se asociaron a cáncer gástrico. Tener una infección por Helicobacter pylori CagA positivo (OR=4,39; IC95% 1,82-10,59) se asoció a la presencia de úlcera duodenal con lesiones preneoplásicas, y ser portador del genotipo IL-1B-511CT se asoció a úlcera duodenal (OR=0,30; IC 95% 0,10-0,91). Conclusión: Los resultados sugieren que la respuesta pro-inflamatoria y la genética virulenta de la bacteria son factores relacionados con los diferentes desenlaces ocasionados por la infección por Helicobacter pylori en la población estudiada; así, el polimorfismo IL-1B-511 es un factor relacionado con cáncer gástrico y úlcera duodenal, y la infección por Helicobacter pylori CagA positivo es un factor asociado a cáncer gástrico y úlcera duodenal con lesiones preneoplásicas.
Objective: To determine the association between the IL-1B-511, IL-1RN, TNF-α-308, IL-10-819 and IL-101082 polymorphisms and positive Heliocobacter pylori CagA infection in a group of patients with gastric cancer and duodenal ulcer in different populations in Colombia. Methods: A case-control study was performed on 341 patients: those with non-atrophic gastritis, 194; with gastric cancer, 58; duodenal ulcer with preneoplastic lesion, 54; and with duodenal ulcer, 35. The genotyping of polymorphisms was done with allelic discrimination using PCR in real time, and that for IL-1RN with conventional PCR and agarose electrophoresis. Helicobacter pylori CagA infection was ascertained with ELISA. Logistic regression was used in statistical analysis. Results: Being a carrier of genotype IL-1B-511TT (OR=4.69; CI 95% 1.22-18.09) and being positive for Helicobacter pylori CagA infection (OR=4.43; CI 95% 1.72-11.4) are associated with gastric cancer. Positive Helicobacter pylori CagA infection (OR=4.39; CI 95% 1.82-10.59) is associated with the presence of duodenal ulcer with preneoplastic lesions, being a carrier of genotype IL-1B-511CT is associated with duodenal ulcer (OR=0.30; CI 95% 0.10-0.91). Conclusion: The results suggest that pro-inflammatory response and virulent bacterial genetics are factors related to the different outcomes brought about by Helicobacter pylori infection in the population studied; that is, the IL-1B-511 polymorphism is a factor related to gastric cancer and duodenal ulcer, and positive Helicobacter pylori CagA infection is a factor associated with gastric cancer and duodenal ulcer with preneoplastic lesions.
Assuntos
Humanos , Adulto , Adenocarcinoma , Adenocarcinoma/classificação , Estudos de Casos e Controles , Helicobacter pylori/classificação , Polimorfismo Genético , Neoplasias Gástricas , Úlcera Duodenal/classificação , Colômbia , Ensaio de Imunoadsorção Enzimática/métodos , Modelos Logísticos , Reação em Cadeia da Polimerase/métodosRESUMO
Background: There is an association of interleukin (IL)1B polymorphism with gastric cancer risk. However systematic reviews of the existing evidence have shown that such association varies across populations with different genetic ancestry. Aim: To evaluate the association of IL-1B-511 and IL-1RN polymorphism and Helicobacter pylori IgG antibodies CagA, with gastric cancer in two Colombian cities located in a high risk area for gastric cancer. Material and Methods: A case-control study including 46 gastric cancer cases and 99 controls with non-atrophic gastritis from a high risk zone for gastric cancer. Polymorphism genotyping was carried out by polymerase chain reaction (PCR) and IgG CagA status by ELISA. Results: IgG CagA seropositive individuals had an increased gastric cancer risk (odds ratio (OR) = 11.56; 95 percent confidence intervals (CI) 2.62-50.91 in Tunja and OR = 19.66, 95 percentCI 0.98-395 in Bogotá). IL-1B-511TT carriers in Tunja had increased risk of gastric cancer (OR = 11.31; 95 percentCI 1.20-106.54)), while IL-1RN*2 alelle carriers in Bogotá showed an inverse association with gastric cancer risk (OR = 0.03; 95 percentCI 0.01-0.65). Conclusions: This study adds evidence to the positive association of Helicobacter pylori CagA positive strains with non-cardial gastric cancer etiology. There is a possible heterogeneity in the association of IL-1B gene polymorphism with cancer, in populations of similar ethnic background and settled in the same risk area.
Assuntos
Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Antígenos de Bactérias/genética , Proteínas de Bactérias/genética , Infecções por Helicobacter/genética , Helicobacter pylori/imunologia , Proteína Antagonista do Receptor de Interleucina 1/genética , Interleucina-1beta/genética , Polimorfismo Genético/genética , Neoplasias Gástricas , Estudos de Casos e Controles , Colômbia/etnologia , Infecções por Helicobacter/imunologia , Helicobacter pylori/genética , Imunoglobulina G/sangue , Fatores de Risco , Neoplasias Gástricas/genética , Neoplasias Gástricas/imunologia , Neoplasias Gástricas/microbiologiaRESUMO
La infección por Helicobacter pylori está asociada con gastritis, úlcera gastroduodenal, linfoma tipo Maltoma, y es considerado un importante factor de riesgo para el desarrollo de cáncer gástrico. El objetivo de este estudio fue caracterizar los genotipos vacA de cepas de H. pylori provenientes de biopsias gástricas de una población venezolana. Se evaluaron 128 pacientes con indicación de endoscopia, por enfermedad de las vías digestivas superiores. Se obtuvieron biopsias gástricas de cada uno de ellos para la amplificación de glm y tipificación de las formas alélicas de vacA, empleando la reacción en cadena de la polimerasa. Los resultados demostraron que 82 (64%) de las muestras fueron positivas para la amplificación de glm. De estos, 51 (62%) de las cepas tenían el genotipo vacA, forma alélica S1 (17 s1a, 29 s1b, 5 s1a+s1b, 10 fueron no tipificables) y 21 (26%) tenían el subtipo S2. El análisis de la región media de vacA reveló que 42 (51%) fueron vac A m1, 26 (32%) m2 y 14 (17%) no tipificaron para m1 o m2. Los resultados de la presente investigación demostraron una alta frecuencia de infección por H pylori genotipo vac A variante alèlica s1/ m1.
Helicobacter pylori infection is associated with gastritis, gastroduodenal ulcer, and Maltoma type lymphoma, and is also considered an important risk factor for the development of gastric cancer. The purpose of this study was to characterize vacA genotypes of H. pylori strains from gastric biopsies from a Venezuelan population. One hundred and twenty eight patients who required endoscopy due to disease of the upper digestive system were evaluated. A gastric biopsy was taken from each of them for glm amplification and typing of the allelic vacA forms, using the polymerase chain reaction assay. Results showed that 82 (64%) of the samples were positive for glm amplification. Of these, 51 (62%) of the strains had the vacA genotype, S1 allelic form (17 sla, 29 slb, 5 sla+slb, 10 were not typable) and 21 (26%) had the S2 subtype. The analysis of the vacA mid region revealed that 42 (51%) were vacA m1, 26 (32%) m2, and 14 did not type for m1 or m2. The results of this investigation showed a high frequency of H. pylori vacA genotype infections, s1/m1 allelic variants.
RESUMO
Helicobacter pylori strains frequently express Lewis X (Le(x)) and/or Le(y) on their cell surfaces as constituents of the O antigens of their lipopolysaccharide molecules. To assess the effect of Le(x) and Le(y) expression on the ability of H. pylori to colonize the mouse stomach and to adhere to epithelial cells, isogenic mutants were created in which fucT1 alone or fucT1 and fucT2, which encode the fucosyl transferases necessary for Le(x) and Le(y) expression, were deleted. C3H/HeJ mice were experimentally challenged with either wild-type 26695 H. pylori or its isogenic mutants. All strains, whether passaged in the laboratory or recovered after mouse passage, colonized the mice well and without consistent differences. During colonization by the mutants, there was no reversion to wild type. Similarly, adherence to AGS and KatoIII cells was unaffected by the mutations. Together, these findings indicate that Le expression is not necessary for mouse gastric colonization or for H. pylori adherence to epithelial cells.
Assuntos
Infecções por Helicobacter/microbiologia , Antígenos do Grupo Sanguíneo de Lewis/fisiologia , Antígenos CD15/fisiologia , Estômago/microbiologia , Animais , Anticorpos Antibacterianos/sangue , Aderência Bacteriana/imunologia , Células Epiteliais/microbiologia , Feminino , Fucosiltransferases/genética , Fucosiltransferases/fisiologia , Infecções por Helicobacter/sangue , Infecções por Helicobacter/imunologia , Helicobacter pylori/enzimologia , Helicobacter pylori/genética , Helicobacter pylori/imunologia , Helicobacter pylori/fisiologia , Mucosa Intestinal/citologia , Camundongos , Camundongos Endogâmicos C3H , MutagêneseRESUMO
La infección por Helicobacter pylori está asociada actualmente con diferentes formas de enfermedades digestivas, incluyendo gastritis, úlceras duodenales, gástricas y linfomas del tipo Maltoma. El microorganismo es considerado como un importante factor de riesgo para el desarrollo de cáncer gástrico. El objetivo de este estudio fue caracterizar la respuesta inmune sérica de Inmunoglobulina G (IgG), contra los antígenos CagA, ureasa y extracto total de Helicobacter pylori, en un grupo de pacientes con diferentes formas de enfermedad de las vías digestivas superiores. Fueron evaluados 66 pacientes sintomáticos referidos para examen endoscópico, provenientes del Servicio de Gastroenterología del Hospital Universitario de Caracas, Venezuela. La respuesta inmune fue evaluada mediante ensayo inmunoabsorbente (ELISA) usando antígenos recombinantes para Ureasa y CagA. La infección por Helicobacter pylori fue determinada por el cultivo de las biopsias gástricas y prueba de ureasa. Los resultados mostraron la presencia de H. pylori en 48/66 (72,7 por ciento) pacientes mediante cultivo microbiológico y prueba de ureasa. De los 66 pacientes evaluados en este estudio, 45 (68 por ciento) fueron positivos para el extracto total de H. pylori, 34/66 (51 por ciento) presentaron anticuerpos contra CagA y 18/66 (27 por ciento), tuvieron anticuerpos anti-ureasa. La tasa de positividad de anticuerpos anti-CagA en pacientes con úlcera gástrica, cáncer gástrico, y gastritis crónica fue de 87,8 por ciento, 77,7 por ciento y 40,8 por ciento respectivamente. Del presente estudio se puede concluir que los niveles de anticuerpos anti-Cag A y anti extracto total fueron similares para pacientes con enfermedad gastroduodenal severa, incluyendo, úlcera gástrica y adenocarcinoma