Identification of Biomarkers for Systemic Distribution of Nanovesicles From Lactobacillus johnsonii N6.2

Harrison, Natalie A.; Gardner, C. L.; Silva, Danilo R da; González, Claudio F.; Lorca, Graciela L.

doi:10.3389/fimmu.2021.723433

Cited by 15 publications

(30 citation statements)

References 67 publications

(101 reference statements)

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“…To identify the mechanism by which NVs reduce apoptosis in βlox5 cells, we performed RNAseq analysis of βlox5 cells treated with NVs from L. johnsonii at 10 8 and 10 10 particles/ml (NV8 and NV10, respectively), in presence or absence of the pool of cytokines. We have recently reported that different proteins and lipids are enriched in NVs when compared to the membranes (MEMB) isolated from L. johnsonii ( 15 ). To compare the differential host response to NVs or MEMB components, βlox5 cells were challenged with 1 or 5 µg/mL MEMB (MEMB1 and MEMB5, respectively) in the presence or absence of the cytokine combination.…”

Section: Resultsmentioning

confidence: 99%

“…Lactobacillus johnsonii was grown in de Man, Rogosa, Sharpe (MRS) media as previously described ( 8 ). NVs and membrane isolation were performed from L. johnsonii cultures grown in exosome-free MRS media ( 15 ). Isolated NVs were immediately quantified using Nanosight NS300 (Malvern instruments Ltd, Malvern, UK) and aliquots were stored at -80°C until use.…”

Section: Methodsmentioning

confidence: 99%

“…One of the limitations of our preclinical and clinical studies has been the absence of identified mechanistic effectors of L. johnsonii activity. As a step in that direction, we have recently characterized extracellular vesicles (herein referred as nanovesicles, NVs) secreted by L. johnsonii ( 15 ). Similar to mammalian cells, both Gram-negative and Gram-positive bacteria are able to release NVs ( 16 ).…”

Section: Introductionmentioning

confidence: 99%

“…These NVs are evolutionarily conserved, have a spherical shape composed of proteolipid bilayers, and carry specific subsets of lipids, nucleic acids, proteins and metabolites ( 17 ). Proteomic and lipidomic analysis showed that NVs secreted by L. johnsonii have a unique protein and lipid composition that is distinct from the L. johnsonii cell membrane ( 15 ). NV surface-exposed proteins were evaluated for their potential to generate an immune response in human hosts.…”

Section: Introductionmentioning

confidence: 99%

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Nanovesicles From Lactobacillus johnsonii N6.2 Reduce Apoptosis in Human Beta Cells by Promoting AHR Translocation and IL10 Secretion

et al. 2022

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L. johnsonii N6.2 releases nano-sized vesicles (NVs) with distinct protein and lipid contents. We hypothesized that these NVs play a central role in the delivery of bioactive molecules that may act as mechanistic effectors in immune modulation. In this report, we observed that addition of NVs to the human pancreatic cell line βlox5 reduced cytokine-induced apoptosis. Through RNAseq analyses, increased expression of CYP1A1, CYP1B1, AHRR, and TIPARP genes in the aryl hydrocarbon receptor (AHR) pathways were found to be significantly induced in presence of NVs. AHR nuclear translocation was confirmed by confocal microscopy. The role of NVs on beta cell function was further evaluated using primary human pancreatic islets. It was found that NVs significantly increased insulin secretion in presence of high glucose concentrations. These increases positively correlated with increased GLUT6 and SREBF1 mRNA and coincided with reduced oxidative stress markers. Furthermore, incubation of NVs with THP-1 macrophages promoted the M2 tolerogenic phenotype through STAT3 activation, expression of AHR-dependent genes and secretion of IL10. Altogether, our findings indicate that bacterial NVs have the potential to modulate glucose homeostasis in the host by directly affecting insulin secretion by islets and through the induction of a tolerogenic immune phenotype.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Nanovesicles From Lactobacillus johnsonii N6.2 Reduce Apoptosis in Human Beta Cells by Promoting AHR Translocation and IL10 Secretion

et al. 2022

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“…On the other hand, it is interesting to note that L. johnsonii N6.2 EVs are recognized by IgA and IgG from the plasma of individuals who had consumed the probiotic. In particular, the increase of IgA occurs as a result of a specific response to EV components: Sdp_SH3b2 and Sdp_SH3b6 ( Harrison et al, 2021 ). Although the function of bacterial SH3b domains is not completely known, they are proposed to be cell wall binding domains in prokaryotes.…”

Section: Extracellular Vesicles From Probiotic Bacteria and Infectiou...mentioning

confidence: 99%

Probiotics, Their Extracellular Vesicles and Infectious Diseases

et al. 2022

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Probiotics have been shown to be effective against infectious diseases in clinical trials, with either intestinal or extraintestinal health benefits. Even though probiotic effects are strain-specific, some “widespread effects” include: pathogen inhibition, enhancement of barrier integrity and regulation of immune responses. The mechanisms involved in the health benefits of probiotics are not completely understood, but these effects can be mediated, at least in part, by probiotic-derived extracellular vesicles (EVs). However, to date, there are no clinical trials examining probiotic-derived EVs health benefits against infectious diseases. There is still a long way to go to bridge the gap between basic research and clinical practice. This review attempts to summarize the current knowledge about EVs released by probiotic bacteria to understand their possible role in the prevention and/or treatment of infectious diseases. A better understanding of the mechanisms whereby EVs package their cargo and the process involved in communication with host cells (inter-kingdom communication), would allow further advances in this field. In addition, we comment on the potential use and missing knowledge of EVs as therapeutic agents (postbiotics) against infectious diseases. Future research on probiotic-derived EVs is needed to open new avenues for the encapsulation of bioactives inside EVs from GRAS (Generally Regarded as Safe) bacteria. This could be a scientific novelty with applications in functional foods and pharmaceutical industries.

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Internalization of extracellular vesicles from Lactobacillus johnsonii N6.2 elicit an RNA sensory response in human pancreatic cell lines

Silva

González

Lorca

2023

J of Extracellular Bio

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Cells of all domains of life can secrete extracellular vesicles (EV). These secreted vesicles have been indicated as vehicles carrying molecules that facilitate intra‐ and inter‐species interaction. Lactobacillus johnsonii N6.2, a bacterium used in probiotic preparations, has been shown to produce nano‐sized EV. In the present work we used L. johnsonii N6.2 EV, concentrated from exosome‐depleted MRS supernatant, to identify the uptake mechanisms of EV and the impact of the RNA cargo in the EV on the upregulation of the cellular response of βlox5 human pancreatic cells. Using eukaryotic uptake inhibitors, it was found that EV are internalized by the clathrin/dynamin mediated endocytosis pathway. Further co‐localization experiments with the endosome markers RAB5, RAB7 and LAMP1 as well as calcein indicated that EV escape the endosome shortly after RAB7 fusion. Using the expression of the 2′,5′‐oligoadenylate synthetase (OAS) host pathway, previously identified as targeted by L. johnsonii EV, we found that the host cellular response to the EV are dependent on the integrity of the external components of the EV as well as on the RNA cargo. Global transcriptome analysis was performed on EV and the bacterial whole cell. It was found that the RNA transcripts found within the EV largely represent the most abundantly transcribed genes in the bacterial cells such as those associated with protein synthesis and glycolysis. Further analysis showed an enrichment of smaller size transcripts as well as those encoding for membrane bound or extracellular proteins in L. johnsonii’s EV.

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Identification of Biomarkers for Systemic Distribution of Nanovesicles From Lactobacillus johnsonii N6.2

Cited by 15 publications

References 67 publications

Nanovesicles From Lactobacillus johnsonii N6.2 Reduce Apoptosis in Human Beta Cells by Promoting AHR Translocation and IL10 Secretion

Nanovesicles From Lactobacillus johnsonii N6.2 Reduce Apoptosis in Human Beta Cells by Promoting AHR Translocation and IL10 Secretion

Probiotics, Their Extracellular Vesicles and Infectious Diseases

Internalization of extracellular vesicles from Lactobacillus johnsonii N6.2 elicit an RNA sensory response in human pancreatic cell lines

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