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EBioMedicine May 2023Probiotics have been increasingly proposed for enhancing immune checkpoint blockade (ICB) treatments against cancer. However, its causal relationship with...
BACKGROUND
Probiotics have been increasingly proposed for enhancing immune checkpoint blockade (ICB) treatments against cancer. However, its causal relationship with immunotherapeutic efficacy remains unclear, which promoted us to explore if and how probiotic Lacticaseibacillus rhamnosus Probio-M9 manipulates gut microbiome for expected outcomes.
METHODS
We evaluated the effects of Probio-M9 on the anti-PD-1 treatment against colorectal cancer in mice via a multi-omics approach. We defined the mechanisms of Probio-M9-mediated antitumor immunity by comprehensive analyses of metagenome and metabolites of commensal gut microbes as well as the immunologic factors and serum metabolome of the host.
FINDINGS
The results indicated that Probio-M9 intervention strengthened the anti-PD-1-based tumor inhibition. Both prophylactic and therapeutic administration of Probio-M9 showed conspicuous performance in controlling tumor growth with ICB treatment. The supplement of Probio-M9 modulated enhanced immunotherapy response through promoting beneficial microbes (e.g., Lactobacillus and Bifidobacterium animalis), producing beneficial metabolites including butyric acids in the gut, and accumulating blood-derived α-ketoglutaric acid, N-acetyl-l-glutamic acid and pyridoxine in particular, which promoted the infiltration and activation of cytotoxic T lymphocytes (CTLs) and suppressing the function of regulatory T cells (Tregs) in the tumor microenvironment (TME). Subsequently, we found that enhanced immunotherapeutic response was transmissible by transplanting either post-probiotic-treatment gut microbes or intestinal metabolites to new tumor-bearing mice.
INTERPRETATION
This study offered valuable insight into the causal role of Probio-M9 in correcting the defects in gut microbiota that compromised anti-PD-1 therapeutic efficacy, which can be used as an alternative synergetic agent with ICB for clinical cancer treatment.
FUNDING
This research was supported by Research Fund for the National Key R&D Program of China (2022YFD2100702), Inner Mongolia Science and Technology Major Projects (2021ZD0014), and China Agriculture Research System of MOF and MARA.
Topics: Animals; Mice; Dietary Supplements; Lacticaseibacillus; Lacticaseibacillus rhamnosus; Neoplasms; Probiotics; Tumor Microenvironment; Immune Checkpoint Inhibitors
PubMed: 37027929
DOI: 10.1016/j.ebiom.2023.104533 -
Journal of Translational Medicine Mar 2023Gut microbiota modulation has been demonstrated to be effective in protecting patients against detrimental effects of anti-cancer therapies, as well as to improve the...
BACKGROUND
Gut microbiota modulation has been demonstrated to be effective in protecting patients against detrimental effects of anti-cancer therapies, as well as to improve the efficacy of certain anti-cancer treatments. Among the most characterized probiotics, Lactobacillus rhamnosus GG (LGG) is currently utilized in clinics to alleviate diarrhea, mucositis or intestinal damage which might be associated with several triggers, including Clostridium difficile infections, inflammatory gut diseases, antibiotic consumption, chemotherapy or radiation therapy. Here, we investigate whether LGG cell-free supernatant (LGG-SN) might exert anti-proliferative activity toward colon cancer and metastatic melanoma cells. Moreover, we assess the potential adjuvant effect of LGG-SN in combination with anti-cancer drugs.
METHODS
LGG-SN alone or in combination with either 5-Fuorouracil and Irinotecan was used to treat human colon and human melanoma cancer cell lines. Dimethylimidazol-diphenyl tetrazolium bromide assay was employed to detect cellular viability. Trypan blue staining, anti-cleaved caspase-3 and anti-total versus anti-cleaved PARP western blots, and annexin V/propidium iodide flow cytometry analyses were used to assess cell death. Flow cytometry measurement of cellular DNA content (with propidium iodide staining) together with qPCR analysis of cyclins expression were used to assess cell cycle.
RESULTS
We demonstrate that LGG-SN is able to selectively reduce the viability of cancer cells in a concentration-dependent way. While LGG-SN does not exert any anti-proliferative activity on control fibroblasts. In cancer cells, the reduction in viability is not associated with apoptosis induction, but with a mitotic arrest in the G2/M phase of cell cycle. Additionally, LGG-SN sensitizes cancer cells to both 5-Fluorouracil and Irinotecan, thereby showing a positive synergistic action.
CONCLUSION
Overall, our results suggest that LGG-SN may contain one or more bioactive molecules with anti-cancer activity which sensitize cancer cells to chemotherapeutic drugs. Thus, LGG could be proposed as an ideal candidate for ground-breaking integrated approaches to be employed in oncology, to reduce chemotherapy-related side effects and overcome resistance or relapse issues, thus ameliorating the therapeutic response in cancer patients.
Topics: Humans; Lacticaseibacillus rhamnosus; Irinotecan; Propidium; Colon; Adjuvants, Immunologic; Melanoma; Probiotics
PubMed: 36918929
DOI: 10.1186/s12967-023-04036-3 -
Frontiers in Cellular and Infection... 2021Bacterial vaginosis (BV) and its recurrence are most commonly associated with the formation of species biofilm. Probiotics are typically used to treat BV; however, the...
Bacterial vaginosis (BV) and its recurrence are most commonly associated with the formation of species biofilm. Probiotics are typically used to treat BV; however, the optimal period of probiotic application in BV treatment remains uncertain. The present study aimed to explore the effects of and on various stages of biofilm formation in species. The biofilm-forming ability of seven strains, including one ATCC 14018 and six clinically isolated species, was determined gentian violet staining assay. Moreover, the sensitivity of the planktonic and biofilm forms toward metronidazole and clindamycin was assessed microdilution broth method. Xbb-LR-1 and Xbb-LC-1 were added during various stages of biofilm formation in species and were cocultured for 24 h. The biofilm thickness of each sample was determined confocal laser scanning microscopy (CLSM). The absolute quantities of species in each sample was obtained real time polymerase chain reaction method, and the pH value was obtained using a pH indicator paper. Biofilm formation by species in a medium with distinct pH values was observed gentian violet staining, CLSM, and scanning electron microscopy (SEM). The biofilm increased the resistance of species toward metronidazole and clindamycin. added at the initial biofilm formation stage in species exhibited highest inhibitory effect, with a percentage inhibition of 38.17% ± 1.35%. When the pH value of the culture medium was <4.5 or >6.5, ATCC 14018 could hardly form a biofilm; however, at pH ≥4.5 and ≤6.5, it was able to form a stronger biofilm. The amount of biofilm attained maximum value at optical density of 3.29 ± 0.28 (595 nm), pH 5.5, and at 36 h. Biofilm formation increases the resistance of species toward antibiotics. Maintaining an acidic vaginal environment with pH <4.5 and a vaginal microbiota dominated by remarkably prevents the formation of species biofilm at the initial stage, which further has a significant impact on the treatment and prevention of biofilm-related infections.
Topics: Biofilms; Female; Gardnerella; Gardnerella vaginalis; Humans; Lacticaseibacillus casei; Lacticaseibacillus rhamnosus; Probiotics; Vagina
PubMed: 33680986
DOI: 10.3389/fcimb.2021.568178 -
Jornal de Pediatria 2015Triggered by the growing knowledge on the link between the intestinal microbiome and human health, the interest in probiotics is ever increasing. The authors aimed to... (Review)
Review
OBJECTIVE
Triggered by the growing knowledge on the link between the intestinal microbiome and human health, the interest in probiotics is ever increasing. The authors aimed to review the recent literature on probiotics, from definitions to clinical benefits, with emphasis on children.
SOURCES
Relevant literature from searches of PubMed, CINAHL, and recent consensus statements were reviewed.
SUMMARY OF THE FINDINGS
While a balanced microbiome is related to health, an imbalanced microbiome or dysbiosis is related to many health problems both within the gastro-intestinal tract, such as diarrhea and inflammatory bowel disease, and outside the gastro-intestinal tract such as obesity and allergy. In this context, a strict regulation of probiotics with health claims is urgent, because the vast majority of these products are commercialized as food (supplements), claiming health benefits that are often not substantiated with clinically relevant evidence. The major indications of probiotics are in the area of the prevention and treatment of gastro-intestinal related disorders, but more data has become available on extra-intestinal indications. At least two published randomized controlled trials with the commercialized probiotic product in the claimed indication are a minimal condition before a claim can be sustained. Today, Lactobacillus rhamnosus GG and Saccharomyces boulardii are the best-studied strains. Although adverse effects have sporadically been reported, these probiotics can be considered as safe.
CONCLUSIONS
Although regulation is improving, more stringent definitions are still required. Evidence of clinical benefit is accumulating, although still missing in many areas. Misuse and use of products that have not been validated constitute potential drawbacks.
Topics: Bifidobacterium; Child; Dermatitis, Atopic; Diarrhea; Dietary Supplements; Gastrointestinal Diseases; Gastrointestinal Tract; Humans; Lacticaseibacillus rhamnosus; Probiotics; Saccharomyces
PubMed: 25458874
DOI: 10.1016/j.jped.2014.08.005 -
Cells Oct 2023Probiotic bacteria belonging to spp. are important producers of bioactive molecules, known as postbiotics, that play essential roles in the immunological support of the...
Probiotic bacteria belonging to spp. are important producers of bioactive molecules, known as postbiotics, that play essential roles in the immunological support of the intestinal mucosa. In this study, the system of co-culture of intestinal epithelial cells with macrophage cells in vitro was used to study the potential effect of postbiotic fractions of and on the modulation of the immune response induced by pro-inflammatory stimuli. This study's results revealed that the presence of probiotic bacterial components on the mucosal surface in the early and late stage of inflammatory conditions is based on cellular interactions that control inflammation and consequent damage to the intestinal epithelium. In our studies, heat killed fractions of probiotic bacteria and their extracted proteins showed a beneficial effect on controlling inflammation, regardless of the strain tested, consequently protecting intestinal barrier damage. In conclusion, the presented results emphasize that the fractions of probiotic bacteria of and may play a significant role in the regulation of LPS-mediated cytotoxic activity in intestinal epithelial cells. The fractions of probiotic strains of and showed the potential to suppress inflammation, effectively activating the anti-inflammatory cytokine IL-10 and modulating the IL-18-related response.
Topics: Humans; Lactobacillus plantarum; Lacticaseibacillus rhamnosus; Lactobacillus; Probiotics; Inflammation
PubMed: 37947616
DOI: 10.3390/cells12212538 -
Applied and Environmental Microbiology Mar 2020GG is one of the most widely marketed and studied probiotic strains. In GG, the gene cluster encodes pili, which are important for some of the probiotic properties of...
GG is one of the most widely marketed and studied probiotic strains. In GG, the gene cluster encodes pili, which are important for some of the probiotic properties of the strain. A previous study showed that the DNA sequence of the gene cluster was not present in some GG variants isolated from liquid dairy products. To examine the stability of the GG genome in an industrial production process, we sequenced the genome of samples of GG (DSM 33156) collected at specific steps of the industrial production process, including the culture collection stock, intermediate fermentations, and final freeze-dried products. We found that the GG genome sequence was unchanged throughout the production process. Consequently, the gene locus was intact and fully conserved in all 31 samples examined. In addition, different production batches of GG exhibited consistent phenotypes, including the presence of pili in final freeze-dried products, and consistent characteristics in assays of probiotic properties. Our data show that GG is highly stable in this industrial production process. GG is one of the best-studied probiotic strains. One of the well-characterized features of the strain is the pili encoded by the gene cluster. These pili are involved in persistence in the gastrointestinal tract and are important for the probiotic properties of GG. Previous studies demonstrated that the GG genome can be unstable under certain conditions and can lose the gene cluster. Since studies have shown that the loss of the gene cluster decreases certain GG probiotic properties, we assessed both the genomic stability and phenotypic properties of GG throughout an industrial production process. We found that neither genomic nor phenotypic changes occurred in the samples. Therefore, we demonstrate that GG retains the cluster and exhibits excellent genomic and phenotypic stability in the specific industrial process examined here.
Topics: Genome, Bacterial; Lacticaseibacillus rhamnosus; Phenotype; Probiotics
PubMed: 31924618
DOI: 10.1128/AEM.02780-19 -
Microbiology Spectrum Aug 2022Cardiac allograft rejection remains a major factor limiting long-term engraftment after transplantation. A novel phosphoinositide 3-kinase (PI3K)/mTOR dual inhibitor,...
Cardiac allograft rejection remains a major factor limiting long-term engraftment after transplantation. A novel phosphoinositide 3-kinase (PI3K)/mTOR dual inhibitor, BEZ235, prolonged cardiac allograft survival by effectively suppressing activation of the PI3K/serine/threonine kinase (AKT)/mTOR pathway. However, long-term usage of pharmacological immunosuppressant drugs can cause intestinal microbiota dysbiosis. We established mouse models of allogeneic heterotopic heart transplantation with different treatments. Fecal samples were collected and subjected to 16S rRNA sequencing and targeted fecal metabolomic analysis. Graft samples were taken for immune cell detection by flow cytometry. Inflammatory cytokines in serum were quantified by enzyme-linked immunosorbent assay (ELISA). Compared to single-target approaches (IC-87114 and rapamycin), BEZ235 more efficiently prolongs cardiac transplant survival. Interestingly, BEZ235 reduces the diversity and abundance of the intestinal microbiota community. We demonstrated that Lactobacillus rhamnosus HN001 rescues the intestinal microbiota imbalance induced by BEZ235. Our data confirmed that the combination of BEZ235 and Lactobacillus rhamnosus HN001 significantly prolongs cardiac transplant survival. A main metabolic product of Lactobacillus rhamnosus HN001, propionic acid (PA), enriches regulatory T (Treg) cells and serves as a potent immunomodulatory supplement to BEZ235. Our study provides a novel and efficient therapeutic strategy for transplant recipients.
Topics: Animals; Dysbiosis; Heart Transplantation; Imidazoles; Lacticaseibacillus rhamnosus; Mice; Phosphoinositide-3 Kinase Inhibitors; Quinolines; RNA, Ribosomal, 16S; TOR Serine-Threonine Kinases
PubMed: 35862958
DOI: 10.1128/spectrum.00794-22 -
Frontiers in Cellular and Infection... 2023Environmental noise exposure is linked to neuroinflammation and imbalance of the gut microbiota. Promoting gut microbiota homeostasis may be a key factor in relieving...
BACKGROUND
Environmental noise exposure is linked to neuroinflammation and imbalance of the gut microbiota. Promoting gut microbiota homeostasis may be a key factor in relieving the deleterious non-auditory effects of noise. This study aimed to investigate the effect of GG (LGG) intervention on noise-induced cognitive deficits and systemic inflammation in rats.
METHODS
Learning and memory were assessed using the Morris water maze, while 16S rRNA sequencing and gas chromatography-mass spectrometry were used to analyze the gut microbiota and short-chain fatty acid (SCFA) content. Endothelial tight junction proteins and serum inflammatory mediators were assessed to explore the underlying pathological mechanisms.
RESULTS
The results indicated that GG intervention ameliorated noise-induced memory deterioration, promoted the proliferation of beneficial bacteria, inhibited the growth of harmful bacteria, improved dysregulation of SCFA-producing bacteria, and regulated SCFA levels. Mechanistically, noise exposure led to a decrease in tight junction proteins in the gut and hippocampus and an increase in serum inflammatory mediators, which were significantly alleviated by GG intervention.
CONCLUSION
Taken together, GG intervention reduced gut bacterial translocation, restored gut and blood-brain barrier functions, and improved gut bacterial balance in rats exposed to chronic noise, thereby protecting against cognitive deficits and systemic inflammation by modulating the gut-brain axis.
Topics: Rats; Animals; Lacticaseibacillus rhamnosus; Brain-Gut Axis; RNA, Ribosomal, 16S; Inflammation; Tight Junction Proteins; Inflammation Mediators; Cognition; Probiotics
PubMed: 37180445
DOI: 10.3389/fcimb.2023.1067367 -
Journal of Microbiology and... Jul 2023Sarcopenia is defined as loss of muscle mass and strength due to aging. Recent studies show that sarcopenia may improve via the gut-muscle axis, suggesting that gut...
Sarcopenia is defined as loss of muscle mass and strength due to aging. Recent studies show that sarcopenia may improve via the gut-muscle axis, suggesting that gut health may affect muscle phenotypes. In this study, we aimed to investigate the ability of JY02 as a probiotic strain isolated from kimchi to alleviate sarcopenia. JY02-conditioned medium (CM) reduced dexamethasone (DEX)-induced myotube diameter atrophy and expression of muscle degradation markers (MuRF1 and atrogin-1) in C2C12 cells. The amelioration of sarcopenia was investigated by measuring body composition (lean mass), hand grip strength, myofibril size (using histological analysis), and mRNA and protein expression of muscle-related factors in a DEX-induced mouse model. The results of these analyses showed that JY02 supplementation promoted the production of muscle-enhancement markers (MHC Iβ, MHC IIα, and Myo-D) and reduced both the production of muscle degradation markers and the symptoms of muscle atrophy (loss of lean mass and muscle strength). We also found decreased levels of pro-inflammatory cytokines (IL-6, IFN- γ) and increased levels of anti-inflammatory cytokines (IL-10) in the serum of DEX+JY02-administered mice compared to those in DEX-treated mice. Overall, these results suggest that JY02 is a potent probiotic supplement that prevents sarcopenia by suppressing muscle atrophy.
Topics: Mice; Animals; Sarcopenia; Lacticaseibacillus rhamnosus; Dexamethasone; Disease Models, Animal; Hand Strength; Muscular Atrophy
PubMed: 36998149
DOI: 10.4014/jmb.2303.03001 -
Applied Microbiology and Biotechnology Jun 2022We report the production and biochemical characterization of an α-carbonic anhydrase (LrhCA) from gram-positive probiotic bacteria Lactobacillus rhamnosus GG. CAs form...
We report the production and biochemical characterization of an α-carbonic anhydrase (LrhCA) from gram-positive probiotic bacteria Lactobacillus rhamnosus GG. CAs form a family of metalloenzymes that catalyze hydration of CO/interconversion between CO and water to bicarbonate ions and protons. They are divided into eight independent gene families (α, β, γ, δ, ζ, η, θ, and ι). Interestingly, many pathogens have been identified with only β- and/or γ-CAs, which can be targeted with CA-specific inhibitors (CAIs) acting as anti-pathogen drugs. Since it is important to study the potential off-target effects of CAIs for both the human body and its commensal bacteria, we took L. rhamnosus GG as our study subject. To date, only a single α-CA has been identified in L. rhamnosus GG, which was successfully produced and biochemically characterized. LrhCA showed moderate catalytic activity with the following kinetic parameters: k of 9.86 × 10 s and k/K of 1.41 × 10 s M. Moderate inhibition was established with 11 of the 39 studied sulfonamides. The best inhibitors were 5-((4-aminophenyl)sulfonamido)-1,3,4-thiadiazole-2-sulfonamide, 4-(2-hydroxymethyl-4-nitrophenyl-sulfonamidoethyl)-benzenesulfonamide, and benzolamide with K values of 319 nM, 378 nM, and 387 nM, respectively. The other compounds showed weaker inhibitory effects. The K of acetazolamide, a classical CAI, was 733 nM. In vitro experiments with acetazolamide showed that it had no significant effect on cell growth in L. rhamnosus GG culture. Several sulfonamides, including acetazolamide, are in use as clinical drugs, making their inhibition data highly relevant to avoid any adverse off-target effects towards the human body and its probiotic organisms. KEY POINTS: • The α-carbonic anhydrase from Lactobacillus rhamnosus GG (LrhCA) is 24.3 kDa. • LrhCA has significant catalytic activity with a kcat of 9.9 × 105 s-1. • Acetazolamide resulted in a marginal inhibitory effect on cell growth.
Topics: Acetazolamide; Carbon Dioxide; Carbonic Anhydrase Inhibitors; Carbonic Anhydrases; Lacticaseibacillus rhamnosus; Sulfonamides
PubMed: 35612631
DOI: 10.1007/s00253-022-11990-3