Nutrient Deficiency Promotes the Entry of Helicobacter pylori Cells into Candida Yeast Cells

Helicobacter pylori, a Gram-negative bacterium, has as a natural niche the human gastric epithelium. This pathogen has been reported to enter into Candida yeast cells; however, factors triggering this endosymbiotic relationship remain unknown. The aim of this work was to evaluate in vitro if variations in nutrient concentration in the cultured medium trigger the internalization of H. pylori within Candida cells. We used H. pylori–Candida co-cultures in Brucella broth supplemented with 1%, 5% or 20% fetal bovine serum or in saline solution. Intra-yeast bacteria-like bodies (BLBs) were observed using optical microscopy, while intra-yeast BLBs were identified as H. pylori using FISH and PCR techniques. Intra-yeast H. pylori (BLBs) viability was confirmed using the LIVE/DEAD BacLight Bacterial Viability kit. Intra-yeast H. pylori was present in all combinations of bacteria–yeast strains co-cultured. However, the percentages of yeast cells harboring bacteria (Y-BLBs) varied according to nutrient concentrations and also were strain-dependent. In conclusion, reduced nutrients stresses H. pylori, promoting its entry into Candida cells. The starvation of both H. pylori and Candida strains reduced the percentages of Y-BLBs, suggesting that starving yeast cells may be less capable of harboring stressed H. pylori cells. Moreover, the endosymbiotic relationship between H. pylori and Candida is dependent on the strains co-cultured.

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Temperatures Outside the Optimal Range for Helicobacter pylori Increase Its Harboring within Candida Yeast Cells

Biology ◽

10.3390/biology10090915 ◽

2021 ◽

Vol 10 (9) ◽

pp. 915

Author(s):

Kimberly Sánchez-Alonzo ◽

Luciano Arellano-Arriagada ◽

Susana Castro-Seriche ◽

Cristian Parra-Sepúlveda ◽

Humberto Bernasconi ◽

...

Keyword(s):

Helicobacter Pylori ◽

Yeast Cells ◽

Rrna Gene ◽

Bacterial Cells ◽

Optimal Range ◽

Viability Assay ◽

Total Dna ◽

Fetal Bovine ◽

H Pylori ◽

Strain Dependent

Helicobacter pylori is capable of entering into yeast, but the factors driving this endosymbiosis remain unknown. This work aimed to determine if temperatures outside the optimal range for H. pylori increase its harboring within Candida. H. pylori strains were co-cultured with Candida strains in Brucella broth supplemented with 5% fetal bovine serum and incubated at 4, 25, 37 or 40 °C. After co-culturing, yeasts containing bacteria-like bodies (Y-BLBs) were observed by optical microscopy, and the bacterium were identified as H. pylori by FISH. The H. pylori 16S rRNA gene was amplified from the total DNA of Y-BLBs. The viability of intra-yeast H. pylori cells was confirmed using a viability assay. All H. pylori strains were capable of entering into all Candida strains assayed. The higher percentages of Y-BLBs are obtained at 40 °C with any of the Candida strains. H pylori also increased its harboring within yeast in co-cultures incubated at 25 °C when compared to those incubated at 37 °C. In conclusion, although H. pylori grew significantly at 40 °C, this temperature increased its harboring within Candida. The endosymbiosis between both microorganisms is strain-dependent and permits bacterial cells to remain viable under the stressing environmental conditions assayed.

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Comparison of CXC Chemokines ENA-78 and Interleukin-8 Expression in Helicobacter pylori-Associated Gastritis

Infection and Immunity ◽

10.1128/iai.69.1.81-88.2001 ◽

2001 ◽

Vol 69 (1) ◽

pp. 81-88 ◽

Cited By ~ 29

Author(s):

Gabriele Rieder ◽

Wolfgang Einsiedl ◽

Rudolf A. Hatz ◽

Manfred Stolte ◽

Georg A. Enders ◽

...

Keyword(s):

Helicobacter Pylori ◽

Protein Expression ◽

Outer Membrane Proteins ◽

Water Soluble ◽

Interleukin 8 ◽

Gastric Epithelium ◽

Mrna Levels ◽

Rt Pcr ◽

H Pylori

ABSTRACT Colonization of the gastric mucosa with Helicobacter pylori is associated with a dense infiltration of granulocytes into the lamina propria in the active phase of gastritis. In this study, we investigated the involvement of epithelial cell-derived neutrophil-activating protein 78 (ENA-78) in development of H. pylori-associated gastritis. Antral biopsies from 27 patients with H. pylori-associated gastritis and 25 from H. pylori-negative individuals were first analyzed for ENA-78 and interleukin-8 (IL-8) mRNA by semiquantitative reverse transcription (RT)-PCR. In H. pylori-positive patients, significantly elevated levels were found for both chemokines (P < 0.05). Only IL-8 mRNA levels differed significantly (P< 0.05) in H. pylori-infected individuals who had serum antibodies for cytotoxin-associated protein CagA versus H. pylori-infected CagA-negative persons. Quantification of ENA-78 transcript levels by competitive RT-PCR yielded a significant 45-fold upregulation for ENA-78 transcripts in biopsies of H. pylori-positive versus H. pylori-negative patients (P < 0.05). In contrast to earlier findings with IL-8, the degree of ENA-78 mRNA upregulation was independent of the grade of activity of gastritis. Immunofluorescence studies on tissues of antral biopsies localized ENA-78 protein expression mainly to the gastric epithelium of H. pylori-positive patients, while control tissues were negative. Upregulation of ENA-78 and IL-8 mRNA and protein expression was also observed in an in vitro system using a gastric adenocarcinoma cell line. Only viable H. pyloriyielded a strong ENA-78 and IL-8 induction, while H. pyloriouter membrane proteins or water-soluble proteins had no significant effect. These data provide evidence for the importance of both IL-8 and ENA-78 in the development and perpetuation of H. pylori-associated gastritis.

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Helicobacter pylori Colonization Drives Urokinase Receptor (uPAR) Expression in Murine Gastric Epithelium During Early Pathogenesis

Microorganisms ◽

10.3390/microorganisms8071019 ◽

2020 ◽

Vol 8 (7) ◽

pp. 1019

Author(s):

Warner Alpízar-Alpízar ◽

Mette E. Skindersoe ◽

Lone Rasmussen ◽

Mette C. Kriegbaum ◽

Ib J. Christensen ◽

...

Keyword(s):

Gastric Cancer ◽

Helicobacter Pylori ◽

Epithelial Cells ◽

Protein Expression ◽

De Novo ◽

Urokinase Receptor ◽

Gastric Epithelium ◽

Upar Expression ◽

H Pylori

(1) Background: Persistent Helicobacter pylori infection is the most important risk factor for gastric cancer. The urokinase receptor (uPAR) is upregulated in lesions harboring cancer invasion and inflammation. Circumstantial evidence tends to correlate H. pylori colonization with increased uPAR expression in the human gastric epithelium, but a direct causative link has not yet been established in vivo; (2) Methods: In a mouse model of H. pylori-induced gastritis, we investigated the temporal emergence of uPAR protein expression in the gastric mucosa in response to H. pylori (SS1 strain) infection; (3) Results: We observed intense uPAR immunoreactivity in foveolar epithelial cells of the gastric corpus due to de novo synthesis, compared to non-infected animals. This uPAR induction represents a very early response, but it increases progressively over time as do infiltrating immune cells. Eradication of H. pylori infection by antimicrobial therapy causes a regression of uPAR expression to its physiological baseline levels. Suppression of the inflammatory response by prostaglandin E2 treatment attenuates uPAR expression. Notwithstanding this relationship, H. pylori does induce uPAR expression in vitro in co-cultures with gastric cancer cell lines; (4) Conclusions: We showed that persistent H. pylori colonization is a necessary event for the emergence of a relatively high uPAR protein expression in murine gastric epithelial cells.

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In vitro transcytosis of Helicobacter pylori histidine-rich protein through gastric epithelial-like cells and the blood–brain barrier

Bioscience Biotechnology and Biochemistry ◽

10.1093/bbb/zbab221 ◽

2021 ◽

Author(s):

Takashi Iwasaki ◽

Aiki Maruyama ◽

Yurika Inui ◽

Toshihiko Sakurai ◽

Tsuyoshi Kawano

Keyword(s):

Alzheimer’S Disease ◽

Alzheimer's Disease ◽

Helicobacter Pylori ◽

Disease Progression ◽

Blood Brain Barrier ◽

Gastric Epithelium ◽

Brain Barrier ◽

Blood Brain ◽

H Pylori

Abstract Recent epidemiological studies have supported the correlation between Helicobacter pylori infection and the development of Alzheimer's disease. HpHpn, a histidine-rich H. pylori protein, forms amyloid-like oligomers; it may be a pathogenic factor for Alzheimer's disease progression. HpHpn may also be transported from the gastric epithelium to the brain. However, HpHpn is secreted from H. pylori on the outer surface of gastric epithelia; therefore, the hypothesized movement of HpHpn across the gastric epithelium to the blood remains controversial. Here, we found the HpHpn showed acidic pH-dependent cellular uptake and subsequent secretion in human gastric epithelial-like carcinoma cells. Furthermore, HpHpn exhibited in vitro permeability across the blood–brain barrier. Although further in vivo experiments are required, our findings suggest that in vitro transcytosis of HpHpn in gastric epithelial cells and the blood–brain barrier may provide new insights into the correlation between H. pylori infections and Alzheimer's disease progression.

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Antagonism of Helicobacter pylori by Bacteriocins of Lactic Acid Bacteria

Journal of Food Protection ◽

10.4315/0362-028x-66.1.3 ◽

2003 ◽

Vol 66 (1) ◽

pp. 3-12 ◽

Cited By ~ 63

Author(s):

TAE-SEOK KIM ◽

JI-WOON HUR ◽

MYEONG-AE YU ◽

CHAN-ICK CHEIGH ◽

KYUNG-NAM KIM ◽

...

Keyword(s):

Antimicrobial Activity ◽

Helicobacter Pylori ◽

Lactic Acid ◽

Lactic Acid Bacteria ◽

Inoculum Size ◽

Strain Atcc ◽

Initial Inoculum ◽

H Pylori ◽

Strain Dependent

Antimicrobial activity of seven bacteriocins produced by lactic acid bacteria against Helicobacter pylori strains (ATCC 43504, Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH [DSM] 4867, DSM 9691, and DSM 10242) was investigated in vitro using a broth microdilution assay. The bacteriocins chosen for the study were nisin A; lacticins A164, BH5, JW3, and NK24; pediocin PO2; and leucocin K. Antimicrobial activity of the bacteriocins varied among the H. pylori strains tested, of which strain ATCC 43504 was the most tolerant. Among the bacteriocins tested, lacticins A164 and BH5 produced by Lactococcus lactis subsp. lactis A164 and L. lactis BH5, respectively, showed the strongest antibacterial activity against H. pylori strains. MICs of the lacticins against H. pylori strains, when assessed by the critical dilution micromethod, ranged from 0.097 to 0.390 mg/liter (DSM strains) or from 12.5 to 25 mg/liter (ATCC 43504), supporting the strain-dependent sensitivity of the pathogen. Pediocin PO2 was less active than the lacticins against four strains of H. pylori, and leucocin K was the least active peptide, with no inhibition toward H. pylori ATCC 43504. Anti-Helicobacter activity of lacticin A164 was dependent on initial inoculum size as well as concentration of the bacteriocin added.

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Human Embryonic Gastric Xenografts in Nude Mice: a New Model of Helicobacter pylori Infection

Infection and Immunity ◽

10.1128/iai.67.4.1798-1805.1999 ◽

1999 ◽

Vol 67 (4) ◽

pp. 1798-1805 ◽

Cited By ~ 12

Author(s):

Alain Lozniewski ◽

Filipe Muhale ◽

Renee Hatier ◽

Armelle Marais ◽

Marie-Christine Conroy ◽

...

Keyword(s):

Helicobacter Pylori ◽

Nude Mice ◽

Ex Vivo ◽

Gastric Epithelium ◽

Rapid Urease Test ◽

Ex Vivo Model ◽

New Model ◽

Urease Test ◽

H Pylori

ABSTRACT In vitro or animal models have been used to investigate the pathogenesis of Helicobacter pylori infection. However, extrapolation to humans of results obtained with these heterologous models remains difficult. We have developed a new model for the study of H. pylori infection that uses human entire embryonic stomachs engrafted in nude mice. At 80 days after implantation, 22 of these xenografts, which exhibited a mature gastric epithelium, were inoculated with 107 to 108 CFU of eitherH. pylori LB1, a freshly isolated H. pyloristrain (n = 12), or H. pylori ATCC 49503 (n = 10). After 12-week examination, H. pylori LB1 persistently colonized the antrum of all inoculated grafts, as assessed by culture (mucus and mucosa), immunohistochemistry (mucosa), and a rapid urease test (mucus). H. pylori ATCC 49503, either before or after in vivo passage, permitted only a transient 2-week colonization in one of the five inoculated grafts in both groups. Colonization was always associated with an increase of gastric juice pH. A mild neutrophil infiltration of the gastric mucosa was noted solely in infected grafts. Transmission electron microscopy showed adherence of H. pylori organisms to epithelial cell surface. In six animals, intracytoplasmic location of this bacterium was observed in the antrum or the fundus. These results allow us to propose this model as a new ex vivo model for the study of specificH. pylori-gastric cell interactions.

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Limited Role of Lipopolysaccharide Lewis Antigens in Adherence of Helicobacter pylori to the Human Gastric Epithelium

Infection and Immunity ◽

10.1128/iai.71.5.2876-2880.2003 ◽

2003 ◽

Vol 71 (5) ◽

pp. 2876-2880 ◽

Cited By ~ 27

Author(s):

Jafar Mahdavi ◽

Thomas Borén ◽

Christina Vandenbroucke-Grauls ◽

Ben J. Appelmelk

Keyword(s):

Helicobacter Pylori ◽

Gastric Epithelium ◽

In Vivo Studies ◽

Minor Role ◽

Human Gastric Mucosa ◽

H Pylori

ABSTRACT In vitro and in vivo studies from various groups have suggested that Helicobacter pylori lipopolysaccharide (LPS) Lewis x (Lex) antigens mediate bacterial adhesion. We have now reevaluated this hypothesis by studying the adherence in situ of H. pylori strain 11637 and its corresponding Lex-negative rfbM mutant to human gastric mucosa from patients (n = 22) with various gastric pathologies. Significant binding of the parent strain was observed in only 8 out of 22 sections; in four out of eight patients, the Lex-negative mutant bound less well. One of these four patients displayed no gastric abnormalities, and the other three showed dysplasia, metaplasia, and adenocarcinoma, respectively; hence, we are unable to define the circumstances under which LPS-mediated adhesion takes place. We conclude that H. pylori LPS plays a distinct but minor role in adhesion.

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Helicobacter pylori Chemotaxis Modulates Inflammation and Bacterium-Gastric Epithelium Interactions in Infected Mice

Infection and Immunity ◽

10.1128/iai.00082-07 ◽

2007 ◽

Vol 75 (8) ◽

pp. 3747-3757 ◽

Cited By ~ 76

Author(s):

Susan M. Williams ◽

Yu-Ting Chen ◽

Tessa M. Andermann ◽

J. Elliot Carter ◽

David J. McGee ◽

...

Keyword(s):

Helicobacter Pylori ◽

Gastric Epithelium ◽

Wild Type ◽

Intimate Association ◽

Directed Motility ◽

Chemotaxis Receptors ◽

H Pylori ◽

Stomach Epithelium

ABSTRACT The ulcer-causing pathogen Helicobacter pylori uses directed motility, or chemotaxis, to both colonize the stomach and promote disease development. Previous work showed that mutants lacking the TlpB chemoreceptor, one of the receptors predicted to drive chemotaxis, led to less inflammation in the gerbil stomach than did the wild type. Here we expanded these findings and examined the effects on inflammation of completely nonchemotactic mutants and mutants lacking each chemoreceptor. Of note, all mutants colonized mice to the same levels as did wild-type H. pylori. Infection by completely nonchemotactic mutants (cheW or cheY) resulted in significantly less inflammation after both 3 and 6 months of infection. Mutants lacking either the TlpA or TlpB H. pylori chemotaxis receptors also had alterations in inflammation severity, while mutants lacking either of the other two chemoreceptors (TlpC and HylB) behaved like the wild type. Fully nonchemotactic and chemoreceptor mutants adhered to cultured gastric epithelial cells and caused cellular release of the chemokine interleukin-8 in vitro similar to the release caused by the wild type. The situation appeared to be different in the stomach. Using silver-stained histological sections, we found that nonchemotactic cheY or cheW mutants were less likely than the wild type to be intimately associated with the cells of the gastric mucosa, although there was not a strict correlation between intimate association and inflammation. Because others have shown that in vivo adherence promotes inflammation, we propose a model in which H. pylori uses chemotaxis to guide it to a productive interaction with the stomach epithelium.

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Helicobacter pylori-induced Rev-erbα fosters gastric bacterial colonization by impairing host innate and adaptive defense

10.21203/rs.3.rs-58644/v1 ◽

2020 ◽

Author(s):

Yuan Zhuang ◽

Fangyuan Mao ◽

Yipin Lv ◽

Chuanjie Hao ◽

Yongsheng Teng ◽

...

Keyword(s):

Helicobacter Pylori ◽

Gastric Mucosa ◽

Bacterial Colonization ◽

Direct Consequence ◽

Gastric Epithelium ◽

Th1 Cell ◽

Extracellular Signal ◽

H Pylori

Abstract Background: Helicobacter pylori (H. pylori) is a human pathogen that infects nearly half of the world’s population, however, the persistent colonization of H. pylori in gastric mucosa remains poorly understood. Nowadays it is believed that impairment of host defense of gastric epithelium induced by H. pylori plays key roles in H. pylori-associated pathology. The nuclear receptor Rev-erbα represents a powerful transcriptional repressor involved in host immunity. However, the regulation, function, and clinical relevance of Rev-erbα in H. pylori infection are presently unknown. Here we demonstrated a pro-colonization role of Rev-erbα in H. pylori infection.Results: Rev-erbα was increased in gastric mucosa of H. pylori-infected patients and mice. H. pylori induced gastric epithelial cells (GECs) to express Rev-erbα via the phosphorylated cagA that activated extracellular signal-regulated kinase (ERK) signaling pathway to mediate transcription factor nuclear factor kappa-B (NF-κB) directly binding to Rev-erbα promoter. Human gastric Rev-erbα expression correlated with H. pylori colonization, and mouse Rev-erbα from non-bone marrow-derived cells promoted gastric H. pylori burden. Importantly, H. pylori colonization was attenuated in Rev-erbα-/- mice and the mice with in vivo pharmacological inhibition of Rev-erbα. Mechanistically, Rev-erbα in GECs not only directly suppressed Reg3b and β-defensin-1 expression via binding to Reg3b and β-defensin-1 promoter respectively, which resulted in impaired bactericidal effects against H. pylori of these antibacterial proteins in vitro and in vivo; but also directly inhibited chemokine CCL21 expression via binding to CCL21 promoter, which led to decreased gastric influx of CD45+CD11c-Ly6G-CD11b+CD68- myeloid cells by CCL21-CCR7-dependent migration and, as a direct consequence, reduced bacterial clearing capacity of H. pylori-specific T helper type 1 (Th1) cell response. Conclusions: Overall, this study identifies a model involving Rev-erbα, which collectively ensures gastric bacterial persistence by suppressing host gene expression required for local innate and adaptive defense against H. pylori, and also highlight a pathological role and an immunosuppressive mechanism of Rev-erbα in persistent H. pylori infection.

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Adherence ofHelicobacter pylorito the Gastric Mucosa

Canadian Journal of Gastroenterology ◽

10.1155/1997/149734 ◽

1997 ◽

Vol 11 (3) ◽

pp. 243-248 ◽

Cited By ~ 7

Author(s):

Marguerite Clyne ◽

Brendan Drumm

Keyword(s):

Helicobacter Pylori ◽

Gastric Mucosa ◽

Bacterial Adhesion ◽

Initial Step ◽

Gastric Epithelium ◽

In Vivo Models ◽

Enteric Diseases ◽

H Pylori

Bacterial adhesion to the intestinal epithelium is a critical initial step in the pathogenesis of many enteric diseases.Helicobacter pyloriis a duodenal pathogen that adheres to the gastric epithelium and causes gastritis and peptic ulceration. The mechanism by whichH pyloricauses disease has not yet been elucidated but adherence to the gastric mucosa is thought to be an important virulence determinant of the organism. What is known about adherence ofH pylorito the gastric mucosa is summarized. Topics discussed are the mechanism ofH pyloriadherence; in vitro and in vivo models ofH pyloriinfection; and adherence and potential adhesins and receptors forH pylori.

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