<i>Lactobacillus johnsonii</i> N6.2 Phospholipids Induce T Cell Anergy upon Cognate Dendritic Cell Interactions

<b>Background/Objectives</b>: <i>Lactobacillus johnsonii</i> N6.2 is a gut symbiont with probiotic properties. <i>L. johnsonii</i> N6.2 delayed the progression of type 1 diabetes (T1D) in diabetic-prone rats. The probiotic intake demonstrated immune cell modulatio...

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Main Authors: Alexandra E. Cuaycal, Monica F. Torrez Lamberti, Graciela L. Lorca, Claudio F. Gonzalez
Format: Article
Language:English
Published: MDPI AG 2025-04-01
Series:Metabolites
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Online Access:https://www.mdpi.com/2218-1989/15/5/284
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author Alexandra E. Cuaycal
Monica F. Torrez Lamberti
Graciela L. Lorca
Claudio F. Gonzalez
author_facet Alexandra E. Cuaycal
Monica F. Torrez Lamberti
Graciela L. Lorca
Claudio F. Gonzalez
author_sort Alexandra E. Cuaycal
collection DOAJ
description <b>Background/Objectives</b>: <i>Lactobacillus johnsonii</i> N6.2 is a gut symbiont with probiotic properties. <i>L. johnsonii</i> N6.2 delayed the progression of type 1 diabetes (T1D) in diabetic-prone rats. The probiotic intake demonstrated immune cell modulation in healthy volunteers, leading to improved wellness and fewer reported symptoms like headaches and abdominal pain. These systemic immune-modulating benefits are attributed to <i>L. johnsonii</i> N6.2’s bioactive fractions, including extracellular vesicles (EVs) and purified phospholipids (PLs). We have previously shown that <i>L. johnsonii</i> N6.2 PLs modulate dendritic cell (DC) function towards a regulatory-like phenotype. Here, we further characterize the immune regulatory effects of <i>L. johnsonii</i> N6.2 PLs on adaptive immunity, specifically upon DC and T cell interactions. We hypothesized that PL-stimulated DCs suppress T cell-mediated responses to maintain tolerance in intra- and extra-intestinal sites. <b>Methods</b>: Bone marrow-derived dendritic cells (BMDCs) were generated from Sprague-Dawley rats and stimulated with <i>L. johnsonii</i> N6.2 PLs. Isogenic T cells were isolated from PBMCs obtained via terminal exsanguination. In vitro cellular assays, co-culture experiments, gene expression analysis by qRT-PCR, and flow cytometry assays were conducted to assess the immune regulatory effects of <i>L. johnsonii</i> N6.2 PLs. <b>Results</b>: The PL-stimulated BMDCs upregulated DC regulatory markers and exhibited an immature-like phenotype with reduced surface expression of maturation markers but increased surface migratory molecules (ICAM-1). These BMDCs presented immunosuppressive functions upon cognate T cell interactions and in the presence of TCR stimulation. Specifically, PL-stimulated BMCDs suppressed Th1 effector function and induced the expression of T cell anergy-related genes after co-culturing for 72 h. <b>Conclusions</b>: This study highlights the immune regulatory capacity of <i>L. johnsonii</i> N6.2’s bioactive components on adaptive immunity, specifically that of purified PLs on DC:T cell-mediated responses leading to immunosuppression. Our findings suggest that <i>L. johnsonii</i> N6.2-purified PLs play a role in regulating adaptive immunity, offering potential benefits for managing immune-related diseases like T1D.
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spelling doaj-art-cf5ca769f67b4adbbcc799a57aadfdab2025-08-20T02:33:47ZengMDPI AGMetabolites2218-19892025-04-0115528410.3390/metabo15050284<i>Lactobacillus johnsonii</i> N6.2 Phospholipids Induce T Cell Anergy upon Cognate Dendritic Cell InteractionsAlexandra E. Cuaycal0Monica F. Torrez Lamberti1Graciela L. Lorca2Claudio F. Gonzalez3Department of Microbiology and Cell Science, Genetics Institute, Institute of Food and Agricultural Sciences, University of Florida, Gainesville, FL 32610, USADepartment of Microbiology and Cell Science, Genetics Institute, Institute of Food and Agricultural Sciences, University of Florida, Gainesville, FL 32610, USADepartment of Microbiology and Cell Science, Genetics Institute, Institute of Food and Agricultural Sciences, University of Florida, Gainesville, FL 32610, USADepartment of Microbiology and Cell Science, Genetics Institute, Institute of Food and Agricultural Sciences, University of Florida, Gainesville, FL 32610, USA<b>Background/Objectives</b>: <i>Lactobacillus johnsonii</i> N6.2 is a gut symbiont with probiotic properties. <i>L. johnsonii</i> N6.2 delayed the progression of type 1 diabetes (T1D) in diabetic-prone rats. The probiotic intake demonstrated immune cell modulation in healthy volunteers, leading to improved wellness and fewer reported symptoms like headaches and abdominal pain. These systemic immune-modulating benefits are attributed to <i>L. johnsonii</i> N6.2’s bioactive fractions, including extracellular vesicles (EVs) and purified phospholipids (PLs). We have previously shown that <i>L. johnsonii</i> N6.2 PLs modulate dendritic cell (DC) function towards a regulatory-like phenotype. Here, we further characterize the immune regulatory effects of <i>L. johnsonii</i> N6.2 PLs on adaptive immunity, specifically upon DC and T cell interactions. We hypothesized that PL-stimulated DCs suppress T cell-mediated responses to maintain tolerance in intra- and extra-intestinal sites. <b>Methods</b>: Bone marrow-derived dendritic cells (BMDCs) were generated from Sprague-Dawley rats and stimulated with <i>L. johnsonii</i> N6.2 PLs. Isogenic T cells were isolated from PBMCs obtained via terminal exsanguination. In vitro cellular assays, co-culture experiments, gene expression analysis by qRT-PCR, and flow cytometry assays were conducted to assess the immune regulatory effects of <i>L. johnsonii</i> N6.2 PLs. <b>Results</b>: The PL-stimulated BMDCs upregulated DC regulatory markers and exhibited an immature-like phenotype with reduced surface expression of maturation markers but increased surface migratory molecules (ICAM-1). These BMDCs presented immunosuppressive functions upon cognate T cell interactions and in the presence of TCR stimulation. Specifically, PL-stimulated BMCDs suppressed Th1 effector function and induced the expression of T cell anergy-related genes after co-culturing for 72 h. <b>Conclusions</b>: This study highlights the immune regulatory capacity of <i>L. johnsonii</i> N6.2’s bioactive components on adaptive immunity, specifically that of purified PLs on DC:T cell-mediated responses leading to immunosuppression. Our findings suggest that <i>L. johnsonii</i> N6.2-purified PLs play a role in regulating adaptive immunity, offering potential benefits for managing immune-related diseases like T1D.https://www.mdpi.com/2218-1989/15/5/284<i>Lactobacillus johnsonii</i> N6.2microbial-derived lipidsphospholipiddendritic cellcellular immune responseimmunomodulation
spellingShingle Alexandra E. Cuaycal
Monica F. Torrez Lamberti
Graciela L. Lorca
Claudio F. Gonzalez
<i>Lactobacillus johnsonii</i> N6.2 Phospholipids Induce T Cell Anergy upon Cognate Dendritic Cell Interactions
Metabolites
<i>Lactobacillus johnsonii</i> N6.2
microbial-derived lipids
phospholipid
dendritic cell
cellular immune response
immunomodulation
title <i>Lactobacillus johnsonii</i> N6.2 Phospholipids Induce T Cell Anergy upon Cognate Dendritic Cell Interactions
title_full <i>Lactobacillus johnsonii</i> N6.2 Phospholipids Induce T Cell Anergy upon Cognate Dendritic Cell Interactions
title_fullStr <i>Lactobacillus johnsonii</i> N6.2 Phospholipids Induce T Cell Anergy upon Cognate Dendritic Cell Interactions
title_full_unstemmed <i>Lactobacillus johnsonii</i> N6.2 Phospholipids Induce T Cell Anergy upon Cognate Dendritic Cell Interactions
title_short <i>Lactobacillus johnsonii</i> N6.2 Phospholipids Induce T Cell Anergy upon Cognate Dendritic Cell Interactions
title_sort i lactobacillus johnsonii i n6 2 phospholipids induce t cell anergy upon cognate dendritic cell interactions
topic <i>Lactobacillus johnsonii</i> N6.2
microbial-derived lipids
phospholipid
dendritic cell
cellular immune response
immunomodulation
url https://www.mdpi.com/2218-1989/15/5/284
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