• Không có kết quả nào được tìm thấy

Dexamethasone inhibits S. aureus- induced neutrophil extracellular Pathogen-Killing Mechanism, Possibly through Toll-like receptor regulation

N/A
N/A
Nguyễn Gia Hào

Academic year: 2023

Chia sẻ "Dexamethasone inhibits S. aureus- induced neutrophil extracellular Pathogen-Killing Mechanism, Possibly through Toll-like receptor regulation"

Copied!
92
0
0

Loading.... (view fulltext now)

Văn bản

In this study, we observed that DXM inhibited bacterial-induced NET formation, but not that induced by PMA. Addition of TLR2 and TLR4 agonists (HKLM and LPS) rescued DXM-inhibited NET formation induced by S.

FigUre 1 | Phorbol 12-myristate 13-acetate (PMa) and Staphylococcus aureus (S. aureus) stimulate neutrophil extracellular traps (neTs) formation  in human neutrophil
FigUre 1 | Phorbol 12-myristate 13-acetate (PMa) and Staphylococcus aureus (S. aureus) stimulate neutrophil extracellular traps (neTs) formation in human neutrophil

Consequently, favorable regulation of the compromised immune homeostasis is effective in the prognosis and treatment of IBD. Amelioration of the symptoms of DSS colitis (exacerbation of severe symptoms, colonic shortening, histopathological changes accompanying tissue damage and myeloperoxidase activation) was observed with oral administration of WT-NCC2705 alone.

FiGURe 1 | A strategy for prevention and treatment of iBD using genetically modified (gm)-immunobiotics
FiGURe 1 | A strategy for prevention and treatment of iBD using genetically modified (gm)-immunobiotics

CONCLUSiON AND FUTURe PeRSPeCTiveS

AUTHOR CONTRiBUTiONS

FUNDiNG

INTRODUCTION

When the dynamic balance between Treg and activated effector T cells is disrupted, homeostasis is compromised and may lead to the development of intestinal mucosal inflammation (Strober et al., 2007). The intersection of these factors may cause an exaggerated pro-inflammatory response against the microbiota that causes inflammatory bowel disease (IBDs), a group of idiopathic and chronic inflammatory disorders of the GIT, mainly comprising ulcerative colitis (CD) and Crohn's disease (UC) (Vangay et al., 2015; Velasquez-Manoff, 2015).

USE OF PROBIOTIC LACTIC ACID BACTERIA IN THE TREATMENT OF

Some LAB microbe-associated molecular patterns (MAMPs) are able to interact with epithelial pattern recognition receptors, mainly the Toll-like receptor-2 (TLR2), TLR6 and nod-like receptors (Ren et al., 2016). This effect has been shown to be sufficient to prevent inflammation in mice (Santos Rocha et al., 2014).

FIGURE 1 | Probiotic LAB anti-inflammatory mechanisms on the intestinal mucosa. (A) Intestinal homeostasis provided by the healthy microbiota role in stimulating ephitelial barrier components such as mucus, Paneth cells activity and eliciting protective im
FIGURE 1 | Probiotic LAB anti-inflammatory mechanisms on the intestinal mucosa. (A) Intestinal homeostasis provided by the healthy microbiota role in stimulating ephitelial barrier components such as mucus, Paneth cells activity and eliciting protective im

THE USE OF RECOMBINANT LACTIC ACID BACTERIA FOR THE TREATMENT

This therapeutic effect was related to the antioxidant properties of recombinant SOD (Rochat et al., 2005). Outside the body, the GMO strain was unable to survive, avoiding its spread in the environment (Steidler et al., 2003).

CONCLUSION

Furthermore, its activation in the intestinal mucosa appears to be required to generate a protective response against the gut microbiota during bacterially driven inflammatory events (Christa et al., 1999; Malka et al., 2000). In addition, the recombinant lactococci that produced PAP improved villous architecture preservation and increased Paneth cell activity in response to 5-FU inflammation (Carvalho et al., 2017).

AUTHOR CONTRIBUTIONS

In fact, the protective effect of PAP in models of GI inflammation has been demonstrated for the first time in a rat model of DSS-induced colitis. Interestingly, after it was shown to be beneficial in the treatment of IBD, another study sought to investigate a protective role of L.

FUNDING

This work used an adenoviral strategy to deliver PAP cDNA into host cells to increase PAP expression (Lv et al personal communication) showed that the use of L.

Oral administration of Lactococcus lactis expressing recombinant 15-lipoxygenase-1 (15 LOX-1) modulates chemically induced colitis in mice.Med. Intragastric administration of a superoxide dismutase-producing recombinant Lactobacillus caseiBL23 strain attenuates DSS colitis in mice.Int.

Bacterial cultures and supplementation

Recently, we have shown that the immune system of Ercc1−/∆7 mice resembles the immune system of aged WT mice. The expression of ERCC1-XPF (excision repair cross-complementation group 1-xeroderma pigmentosum group F) DNA repair endonuclease is reduced to ~5% compared to Ercc1+/+ mice.

Based on a variety of histological, functional, metabolomic, and proteomic data, it was concluded that Ercc1-/∆7 mice resemble normal murine aging ( 22 ). BM, thymus, spleen, mesenteric lymph nodes (MLN) and Peyer's patches (PPs) were isolated. plantarum prevented the age-related decline in colonic mucosal barrier of accelerated aging Ercc1−/∆7 mice.

MiT-chips/16s sequencing

For the detection of bacteria, tissue sections were used for fluorescence in situ hybridization (FISH), as previously published (8).

The Mucus layer in the colon Declines with age

Bacterial supplementations Do not change the Mucus layer in colon

Redundancy analysis of the microbial composition after bacterial supplementation, at the genus-like level of the MIChip analysis. The first and second ordination axes are plotted, showing respectively 4.9 and 3.6% of the variability in the data set.

Bacterial supplementation associated with Minor alterations in colonic

Eubacterium plexicaudatum and a close relative of Anaerostipes caccae tended to be higher (p = 0.06) in Ercc1−/∆7 mice supplemented with L. These data show some differences in microbial species between control-treated Ercc1−/∆7 mice and Ercc1−/∆7 bacteria supplement-treated.

Distinct gene expression Profiles in colon after each Bacterial supplementation

Bacterial supplementation alters growth- and immune-related Pathways in colon

Venn diagram of the total number of upregulated and downregulated genes in the proximal colon of Ercc1−/Δ7 mice treated with LP, BB, or LC (p < 0.05 and >1.2-fold change). In WT mice, we found no effect of bacterial supplementation on the distribution of immune cells in the BM or thymus, except for B-lineage cells after L.

FigUre 4 | Venn diagrams of differentially regulated genes in colon of  Ercc1 −/ ∆ 7  mice after bacterial supplementations
FigUre 4 | Venn diagrams of differentially regulated genes in colon of Ercc1 −/ ∆ 7 mice after bacterial supplementations

Bacterial supplementations Do not alter lifespan of Ercc1 −/∆7 Mice

CD11b + Ly6G-CD68 + cells were divided into Ly6Chi, Ly6Cint and Ly6Clo monocytes. e–g) Mean frequencies of Ly6Chimonocytes, neutrophils and CD3+CD4+CD8−Rorγt+ Th17 cells were determined by flow cytometry. h) Mean concentration of IL-17A production by splenocytes stimulated with ConA for 4 days, as determined by Cytometric Bead Array. Granulocyte monocyte precursors (GMP) were defined as. e–h) Mean absolute numbers were determined by cell counts and flow cytometry.

DiscUssiOn

Data represent mean + SEM from three to six animals per group. Plantarum supplementation altered myeloid and lymphoid development in the bone marrow and thymus of Ercc1-/∆7 mice. a. The effects of the candidate probiotic strains were pronounced on the mucosal barrier in the colon of Ercc1−/∆7 mice compared to WT mice.

Improvement of immunity in the elderly by dietary supplementation with the probiotic Bifidobacterium lactis HN019. Development and application of the human intestinal tract chip, a phylogenetic microarray: analysis of universally conserved phylotypes in the abundant microbiota of young and older adults.

Bene K, Varga Z, Petrov VO, Boyko N and Rajnavolgyi E (2017) Gut microbiota species can elicit both inflammatory and tolerogenic immune responses in human dendritic cells mediated by retinoic acid receptor alpha ligation. In humans, the vitamin A derivative all-trans retinoic acid (ATRA) is endogenously produced from retinol by DCs, macrophages, and epithelial and stromal cells and binds to RARα and retinoic X receptor alpha (RXRα) with different affinities (28) and allows monitoring of the modulatory effects of the retinoid-responsive agents.

ATRA, the selective RARα antagonist BMS-195614 (BMS614), the vehicle dimethyl sulfoxide (DMSO), and the anti-hβ-actin mAb were from Sigma-Aldrich, Schnelldorf, Germany. The anti-hIRF4 antibody was from Cell Signaling Technology, Inc. Trask Lane, Danvers, MA, USA).

Bacterial growth for moDc activation

Phagocytosis assay

Flow cytometry

Measurement of cytokine concentration

To detect IL-17 secretion, plates were coated with 0.5 µg/ml mouse anti-hCD3 antibody (BD Biosciences). Naïve CD4 + T lymphocytes were isolated with the naive CD4 + T Cell Isolation Kit II, human (Miltenyi Biotec).

To detect regulatory T lymphocytes, activated and resting moDCs were washed and co-cultured with PBL or naive CD4+ T lymphocytes for 6 days in serum-free AIM-V medium at a moDC:T cell ratio of 1:10. To detect the presence of intracellular IL-10, T cells were treated with Golgi-Stop™ on day 6.

Western Blotting

Plates were analyzed using an ImmunoScan plate reader (Cell Technology Limited, Shaker Heights, OH, USA). On day 6, cells were harvested, permeabilized, and fixed with the Citofix/Cytoperm intracellular staining kit (BD Biosciences).

Transcription Factors and the cell surface expression of cD1 glycoprotein

The relative gene expression levels of retinoic acid receptor alpha (RARα), retinoic X receptor alpha (RXRα), peroxisome proliferator-activated receptor gamma (PPARγ), ALDH1A2 and CD1d (a), interferon regulatory factor (IRF)4 (F) and IRF8 (g) were measured by quantitative real-time PCR, and the cell surface expression level of CD1a, CD1d (B ), DC-SIGN (c), CD1 4 (D) and CD11b (e) were measured by flow cytometry. The mean values ​​of the relative mRNA levels and the ratio of positive moDCs for the measured cell surface proteins were calculated from five independent experiments + SD.

Differently as compared to rar α hi irF4 lo cells

The T cell polarization capacity of moDCs was monitored in moDC stimulated with Escherichia coli Schaedler, Morganella Morganii and Bacillus subtilis or lipopolysaccharide (LPS) followed by co-cultivation of the cells with freshly isolated autologous T cells for 4 days. Using the in vitro system, we found that the live commensal bacteria were able to upregulate the cell surface expression of CX3CR1 within 24 h (Figure 3B,C), but had no effect on CD103.

The Phagocytic capacity of moDcs

The number of cytokine-producing T lymphocytes stimulated by LPS or by moDC exposed to commensal bacteria was measured by ImmunoSpot enzyme-linked interferon gamma (IFNγ) (a) and interleukin (IL)-17 (B) assays. ANOVA followed by Bonferroni's multiple comparison tests was used in statistical analysis with significance defined as *P < 0.05, **P < 0.01 and.

Depends on the individual characteristics of the Tested Bacteria and on actual

These results indicated that lipid antigen presentation by moDCs via CD1a and CD1d proteins is regulated by both ATRA and the gut microbiota in a species-specific manner.

Polarizing capacity of moDcs but aTra interferes with This effect

In addition, we can rule out the effects of other RAR isoforms such as RARβ, as it is not expressed and the expression of RARβ could not be induced in moDCs in the presence of ATRA. All-trans retinoic acid (ATRA) modifies the differentiation of moDCs that could be prevented by selective inhibition of RARα. a) In the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-4, monocytes differentiate into CD1d-CD1a+ DCs (43).

FigUre 4 | characteristics of the inflammatory and migratory potential of monocyte-derived dentritic cell (moDc) populations induced by  commensal bacteria
FigUre 4 | characteristics of the inflammatory and migratory potential of monocyte-derived dentritic cell (moDc) populations induced by commensal bacteria

PPARgamma controls CD1d expression by activating retinoic acid synthesis in developing human dendritic cells. Small intestinal lamina propria dendritic cells induce de novo generation of Foxp3 T reg cells via retinoic acid.

Treatment and induction of experimental colitis Using TnBs

Although lactate has been known to biochemists for more than 200 years, it has been considered a simple carbon metabolite intermediate with specific organoleptic/antimicrobial properties rather than a bioactive molecule. Recently, lactate has been rediscovered as an active signaling metabolite in numerous areas of biology and medicine (14).

The metabolic outcome of gut microbiota modification is the production of different profiles of short-chain fatty acids (SCFAs), such as butyrate, propionate, and acetate, and these metabolites are important in modulating key signaling pathways involved in gastrointestinal mucosal inflammation (7–9). The influence of probiotic bacteria on intestinal health has been investigated with the aim of preventing IBD or improving its treatment (10-12), and it has also been shown that metabolites present in the supernatants of fermented milk products can ex vivo have a protective effect on the intestinal mucosa exposed to inflammatory processes (13).

To determine whether the immunomodulatory capacity of lactate works in vivo, the present work evaluated the effect of lactate in congenitally driven murine model of colitis. Male BALB/c AnN, 6-week-old mice weighing over 20 g were purchased from the Faculty of Science Veterinary of the National University of La Plata, Argentina.

These suspensions were homogenized, enriched with total viable bacteria by incubation for 24 h at 37 °C, and used to inoculate BHI agar plates. Bacterial translocation was determined by growth of the microorganism on the plates after 48–72 hours of incubation at 37 °C.

Results from one representative experiment out of five are shown, expressed as mean ± SD, *indicates significant difference at p < 0.05 from its corresponding control. Results from one representative experiment out of five are shown. a) Histopathological activity index assigned to different experimental groups.

We observed that Caco-2 intestinal epithelial cells decreased their rate of glucose consumption in the presence of lactate under both basal and flagellin conditions. As epithelial cells may also contribute to increased inflammation in the TNBS model, this could be another possible cellular target explaining the bioactive properties of lactate.

TaBle 1 | Microbial translocation to liver observed 48 h after  2,4,6-trinitrobenzenesulfonic acid (TnBs)-induced colitis.
TaBle 1 | Microbial translocation to liver observed 48 h after 2,4,6-trinitrobenzenesulfonic acid (TnBs)-induced colitis.

Overall, these findings support a link between gut microbial ecology and host metabolism in the pathological setting of SLE. Epidemiological studies have consistently shown an increase in the prevalence and severity of many metabolic disorders in patients with systemic lupus erythematosus (SLE) compared with the general population (1–3).

Patients and controls

In fact, the gut microbiota is seen by some authors as a distinct endocrine organ involved, through a molecular cross-talk with the host, in the maintenance of energy homeostasis and fat storage (19). Thus, it can be speculated that changes in gut microbial ecology may disrupt this homeostatic cross-talk and accelerate the development of pathological outcomes in the host.

Quantification of Total FFa serum levels

Currently, extensive research efforts are focused on deciphering the basis of the cross-talk between the microbiota and the host metabolism in the development and progression of host diseases and have revealed the relevance of the gut microbiota-host metabolism axis mediated by different bacterial and host metabolites (20, 21). All participants were informed and gave a signed informed consent before being included in the study.

Correlations were assessed by Spearman rank tests (r-coefficient and p-value are indicated for each parameter). Differences in demographic and blood lipid variables were assessed by Mann-Withney U tests, while differences in daily intake were analyzed by multivariate analyzes adjusted for confounders.

Energy was adjusted for gender and age, while the rest of the nutrients were adjusted for gender, age and energy.

Total FFa serum levels in sle Patients

The associations between serum FFA levels and abundance of microbial groups at the level of phyla in HC and SLE patients were analyzed by Spearman rank test (r coefficient and p value are indicated for each parameter). To this end, the associations between FFA levels and the main gut microbial groups analyzed by a metagenomic approach as already described (22) were assessed.

Therefore, parameters other than those indicated may explain the altered serum FFA levels registered in SLE. The association between serum FFA levels and the F/B ratio was analyzed by Spearman's rank correlation tests.

FFa Profiling in sle Patients

Additional factors, such as disease status, may also influence the outcome of associations between gut microbiota composition and SCFA-FFA interaction. Notably, F/B ratio was positively associated with PC2 score (Table 8) in HC but not in SLE, thus suggesting a beneficial effect of gut microbiota composition on serum FFA pool in healthy individuals.

FFa and serum Biomarkers in sle Patients

The association between the F/B ratio and serum FFA levels highlights the ability of the gut microbiota to promote systemic responses in the host. Furthermore, we have provided proof-of-concept evidence for the involvement of gut dysbiosis in metabolic changes in lupus.

TaBle 7 | specific free fatty acids (FFa) in healthy controls (hc) and  systemic lupus erythematosus (sle) patients.
TaBle 7 | specific free fatty acids (FFa) in healthy controls (hc) and systemic lupus erythematosus (sle) patients.

FUnDing

The fact that no change in gut microbiota was associated with disease duration (despite the wide range of disease durations studied in the present report) is also consistent with this idea, perhaps suggesting that gut dysbiosis may be present in the preclinical stages of the disease. Larsen N, Vogensen FK, van den Berg FWJ, Nielsen DS, Andreasen AS, Pedersen BK, et al.

BaCKGRoUND

1 Gut Microbiome Research Group, Korea Food Research Institute, Sungnam, South Korea, 2 Department of Food Biotechnology, Korea University of Science and Technology, Daejeon, South Korea, 3 Research and Development Center, Cell Biotech Co. Ltd., Gimpo, South Korea, 4 Department of Biology, Probiotic Kyung Hee University, Seoul. , genome sequence, pacBio.

MateRIaLS aND MetHoDS

Kang J, Chung W-H, Lim T-J, Whon TW, Lim S ken Nam Y-D (2017) Kompleto a panagsasaruno ti genome ti Lactobacillus casei LC5, ti potensial a probiotiko para iti atopic dermatitis.

Bacterial Strains and DNa preparation

Genome Sequencing, assembly, and annotation

ReSULtS

Genome Characteristics of L. casei LC5

Comparative Study of L. casei Group

Functional Classification

The probiotic bacterium Lactobacillus casei induces activation of the intestinal mucosal immune system through innate immunity. Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees.

FIGURe 1 | Genome characteristics of Lactobacillus casei LC5 genome. (a) Circular map of genomic features; eight tracks were plotted in the map
FIGURe 1 | Genome characteristics of Lactobacillus casei LC5 genome. (a) Circular map of genomic features; eight tracks were plotted in the map

Data aCCeSS

Hình ảnh

FiGURe 1 | A strategy for prevention and treatment of iBD using genetically modified (gm)-immunobiotics
FIGURE 1 | Probiotic LAB anti-inflammatory mechanisms on the intestinal mucosa. (A) Intestinal homeostasis provided by the healthy microbiota role in stimulating ephitelial barrier components such as mucus, Paneth cells activity and eliciting protective im
TABLE 1 | Heterolgous proteins with anti-inflammatory properties produced in different strains of lactic acid bacteria.
FigUre 1 | Treatment with L. plantarum prevented the age-related decline in colonic mucus barrier of accelerated aging Ercc1 −/ ∆ 7  mice
+7

Tài liệu tham khảo

Tài liệu liên quan

* Activity 1: Listen and repeat -T introduces sound : / ch / -T asks Ss to listen to the cassette -Ss: repeat chorally – individually -T introduces word: kithchen -T asks