Molecular identification of potential Th1/Th2 responses-modulating bacterial genes using suppression subtractive DNA hybridization

Ghadimi, Darab; Njeru, Patrisio Njiru; Guigas, Claudia; Hassan, M.; Fölster‐Holst, Regina; Geis, Arnold; Vrese, Michael de; Schrezenmeir, Jürgen; Heller, Knut J.

doi:10.1016/j.imbio.2013.10.005

Cited by 9 publications

(10 citation statements)

References 69 publications

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“…For example, the EPS-deficient mutant of L. casei Shirota stimulates more proinflammatory cytokine production in mouse macrophages and spleen cells than in the parental strain (18). Ghadimi et al also suggested that the presence of genes to synthesize polysaccharide might potentially modulate the immunomodulatory property of Lactobacillus fermentum strains toward Th1-inducing cytokine secretion in PBMCs, while the strain lacks the genes that lead to Th1/Th2-suppressing cytokine production (19). Interestingly, L. plantarum strain SF2A35B was shown previously to elicit very limited immune responses in PBMCs and blood monocyte-derived dendritic cells (34,58).…”

Section: Discussionmentioning

confidence: 99%

“…EPS, specifically heteropolysaccharides, of lactobacilli have been reported to be involved in various biological functions, such as phage absorption (14,15), adhesion to human cells or to other bacteria (16,17), and immunomodulation (18,19). Various studies have employed gene deletion mutation of EPS-associated genes as a strategy to investigate the biological roles of EPS.…”

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confidence: 99%

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Strain-Specific Features of Extracellular Polysaccharides and Their Impact on Lactobacillus plantarum-Host Interactions

Lee

Caggianiello

Swam

et al. 2016

Appl Environ Microbiol

110

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Lactobacilli are found in diverse environments and are widely applied as probiotic, health-promoting food supplements. Polysaccharides are ubiquitously present on the cell surface of lactobacilli and are considered to contribute to the species-and strainspecific probiotic effects that are typically observed. Two Lactobacillus plantarum strains, SF2A35B and Lp90, have an obvious ropy phenotype, implying high extracellular polysaccharide (EPS) production levels. In this work, we set out to identify the genes involved in EPS production in these L. plantarum strains and to demonstrate their role in EPS production by gene deletion analysis. A model L. plantarum strain, WCFS1, and its previously constructed derivative that produced reduced levels of EPS were included as reference strains. The constructed EPS-reduced derivatives were analyzed for the abundance and sugar compositions of their EPS, revealing cps2-like gene clusters in SF2A35B and Lp90 responsible for major EPS production. Moreover, these mutant strains were tested for phenotypic characteristics that are of relevance for their capacity to interact with the host epithelium in the intestinal tract, including bacterial surface properties as well as survival under the stress conditions encountered in the gastrointestinal tract (acid and bile stress). In addition, the Toll-like receptor 2 (TLR2) signaling and immunomodulatory capacities of the EPS-negative derivatives and their respective wild-type strains were compared, revealing strain-specific impacts of EPS on the immunomodulatory properties. Taken together, these experiments illustrate the importance of EPS in L. plantarum strains as a strain-specific determinant in host interaction. IMPORTANCEThis study evaluates the role of extracellular polysaccharides that are produced by different strains of Lactobacillus plantarum in the determination of the cell surface properties of these bacteria and their capacity to interact with their environment, including their signaling to human host cells. The results clearly show that the consequences of removal of these polysaccharides are very strain specific, illustrating the diverse and unpredictable roles of these polysaccharides in the environmental interactions of these bacterial strains. In the context of the use of lactobacilli as health-promoting probiotic organisms, this study exemplifies the importance of strain specificity.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Strain-Specific Features of Extracellular Polysaccharides and Their Impact on Lactobacillus plantarum-Host Interactions

Lee

Caggianiello

Swam

et al. 2016

Appl Environ Microbiol

110

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“…PBMCs were cultured in 24-well plated (Corning Incorporated Co., Ltd., China) with final concentration of 1 × 10 6 cells per mL in the presence or absence of different stimulants at 37 °C in 5% CO 2 . Final concentration of 1 × 10 6 CFU mL −1 live-bacterial cell suspensions were used as stimulants, Escherichia coli O111: B4 LPS (Sigma, USA) and concanavalian A (ConA, Shanghai Yuanye Biotechnology Co., Ltd., China) from Canavalia A were used to stimulate PBMCs at a concentration of 1 and 5 μg mL −1 respectively as positive controls, [12,13] PBMCs without stimulation were used as control. The supernatants were collected at 48 h by centrifugation (3000 × g, 15 min) from co-culture.…”

Section: Stimulation Of Pbmcsmentioning

confidence: 99%

Probiotics Alleviate Food Protein Allergy in Mice by Activating TLR4 Signaling Pathway

Tian

Liu

et al. 2023

Molecular Nutrition Food Res

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Scope Food allergy has become a world recognized public health problem due to its versatility and lack of efficacious methods for its treatment. Probiotics supplement is a potential way to prevent food allergy. Methods and results In this study, potential strains are screen out by peripheral blood mononuclear cells (PBMCs), and their abilities of alleviating food allergy are examined using a mouse model induced by ovalbumin (OVA). The results show that six strains increase ratio of interferon‐γ (IFN‐γ)/interleukin (IL)‐4 secreted by PBMCs with good abilities in intestinal adhesion and gastrointestinal tolerance. Oral administration of Bifidobacterium animalis KV9 (KV9) and Lactobacillus vaginalis FN3 (FN3) attenuates allergic responses in allergy mice, including allergic symptoms, mast cells aggregation and activity, serum OVA‐special‐immunoglobulins E (OVA‐sIgE) production. KV9 and FN3 upregulate the production of IFN‐γ/IL‐4 in splenocytes, increase the genes and proteins expression of toll‐like receptor 4 (TLR4), myeloid differentiation primary response 88 (Myd88) and interferon regulatory factor (IRF)‐1 in allergic mice spleen, and decrease the IRF‐4. Conclusion The study demonstrates that KV9 and FN3 possess anti‐allergic activities via activation of TLR4 pathway and modulating the expression of IRF‐1 and IRF‐4 which leads to T helper type 1 (Th1)/T helper type 2 (Th2) cell immunology balance.

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“…Studies on the analysis of gene expression and differentiation in amoebas isolated from different clinical cases may contribute to a better understanding of the parasite's biology. There are many methodologies used with this purpose, and new techniques of differential gene expression have been described [ 6 , 9 – 12 ]. Nevertheless, the suppression subtractive hybridization (SSH) technique, which is based on the identification of genes expressed only in cells and tissues of interest, still has not been used in order to identify genes which are possibly involved in the virulence of amoebas.…”

Section: Introductionmentioning

confidence: 99%

“…Nevertheless, the suppression subtractive hybridization (SSH) technique, which is based on the identification of genes expressed only in cells and tissues of interest, still has not been used in order to identify genes which are possibly involved in the virulence of amoebas. SSH combines cDNA hybridizations with polymerase chain reaction (PCR) and enables high level-expressed genes not to be cloned and genes which are specific to certain structures, generally low expressed, to be selected [ 9 , 13 ], contributing to characterize novel genes which may determine virulence, resistance to drugs, and infectivity. SSH has been used in order to evaluate differential gene expression between two E. histolytica strains, one virulent and another nonvirulent.…”

Section: Introductionmentioning

confidence: 99%

Differentially Expressed Genes of Virulent and NonvirulentEntamoeba histolyticaStrains Identified by Suppression Subtractive Hybridization

Freitas

Alvarenga

Fernandes

et al. 2014

BioMed Research International

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Entamoeba histolytica is a parasite which presents capacity to degrade tissues and therefore has a pathogenic behavior. As this behavior is not shown by all strains, there have been several studies investigating molecular basis of the cytotoxicity process. Using the suppression subtractive hybridization (SSH) technique, differential gene expressions of two E. histolytica strains, one virulent (EGG) and one nonvirulent (452), have been analyzed with the purpose of isolating genes which may be involved with amoebic virulence. Nine cDNA fragments presenting high homology with E. histolytica previously sequenced genes were subtracted. Of these, four genes were confirmed by RT-PCR. Two coding for hypothetical proteins, one for a cysteine-rich protein, expressed only in the virulent strain, EGG and another one, coding for grainin 2 protein, exclusive from 452 strain. This study provided new insight into the proteins differences in the virulent and nonvirulent E. histolytica strains. We believe that further studies with these proteins may prove association of them with tissue damage, providing new perceptions to improve treatment or diagnosis of the invasive disease.

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Molecular identification of potential Th1/Th2 responses-modulating bacterial genes using suppression subtractive DNA hybridization

Cited by 9 publications

References 69 publications

Strain-Specific Features of Extracellular Polysaccharides and Their Impact on Lactobacillus plantarum-Host Interactions

Strain-Specific Features of Extracellular Polysaccharides and Their Impact on Lactobacillus plantarum-Host Interactions

Probiotics Alleviate Food Protein Allergy in Mice by Activating TLR4 Signaling Pathway

Differentially Expressed Genes of Virulent and NonvirulentEntamoeba histolyticaStrains Identified by Suppression Subtractive Hybridization

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