Surface layer of Lactobacillus helveticus MIMLh5 promotes endocytosis by dendritic cells

Valentina Taverniti; Mauro Marengo; Eva Fuglsang; Helene Marie Skovsted; Stefania Arioli; Giacomo Mantegazza; Giorgio Gargari; Stefania Iametti; Francesco Bonomi; Simone Guglielmetti; Hanne Frøkiær

doi:10.1128/aem.00138-19

Surface layer of Lactobacillus helveticus MIMLh5 promotes endocytosis by dendritic cells

Valentina Taverniti^*, Mauro Marengo, Eva Fuglsang, Helene Marie Skovsted, Stefania Arioli, Giacomo Mantegazza, Giorgio Gargari, Stefania Iametti, Francesco Bonomi, Simone Guglielmetti, Hanne Frøkiær

^*Corresponding author for this work

1 Citation (Scopus)

Abstract

Surface layers (S-layers) are proteinaceous arrays covering the cell walls of numerous bacteria. Their suggested properties, such as interactions with the host immune system, have been only poorly described. Here, we aimed to elucidate the role of the S-layer from the probiotic bacterial strain Lactobacillus helveticus MIMLh5 in the stimulation of murine bone-marrow-derived dendritic cells (DCs). MIMLh5 induced greater production of interferon beta (IFN-β), interleukin 10 (IL-10), and IL- 12p70, compared to S-layer-depleted MIMLh5 (naked MIMLh5 [n-MIMLh5]), whereas the isolated S-layer was a poor immunostimulator. No differences in the production of tumor necrosis factor alpha (TNF-α) or IL-1β were found. Inhibition of the mitogen-activated protein kinases JNK1/2, p38, and ERK1/2 modified IL-12p70 production similarly in MIMLh5 and n-MIMLh5, suggesting the induction of the same signaling pathways by the two bacterial preparations. Treatment of DCs with cytochalasin D to inhibit endocytosis before the addition of fluorescently labeled MIMLh5 cells led to a dramatic reduction in the proportion of fluorescencepositive DCs and decreased IL-12 production. Endocytosis and IL-12 production were only marginally affected by cytochalasin D pretreatment when fluorescently labeled n-MIMLh5 was used. Treatment of DCs with fluorescently labeled S-layercoated polystyrene beads (Sl-beads) resulted in much greater uptake of beads, compared to noncoated beads. Prestimulation of DCs with cytochalasin D reduced the uptake of Sl-beads more than plain beads. These findings indicate that the S-layer plays a role in the endocytosis of MIMLh5 by DCs. In conclusion, this study provides evidence that the S-layer of L. helveticus MIMLh5 is involved in endocytosis of the bacterium, which is important for strong Th1-inducing cytokine production.

Original language	English
Article number	e00138-19
Journal	Applied and Environmental Microbiology
Volume	85
Issue number	9
ISSN	0099-2240
DOIs	https://doi.org/10.1128/aem.00138-19
Publication status	Published - 2019

Keywords

Cytochalasin D
Cytokines
MAPKs
Nanoparticles
Probiotic

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10.1128/aem.00138-19

https://aem.asm.org/content/aem/85/9/e00138-19.full.pdf

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@article{dd3f8b69167f4cb39fd161d7b49a7d40,

title = "Surface layer of Lactobacillus helveticus MIMLh5 promotes endocytosis by dendritic cells",

abstract = "Surface layers (S-layers) are proteinaceous arrays covering the cell walls of numerous bacteria. Their suggested properties, such as interactions with the host immune system, have been only poorly described. Here, we aimed to elucidate the role of the S-layer from the probiotic bacterial strain Lactobacillus helveticus MIMLh5 in the stimulation of murine bone-marrow-derived dendritic cells (DCs). MIMLh5 induced greater production of interferon beta (IFN-β), interleukin 10 (IL-10), and IL- 12p70, compared to S-layer-depleted MIMLh5 (naked MIMLh5 [n-MIMLh5]), whereas the isolated S-layer was a poor immunostimulator. No differences in the production of tumor necrosis factor alpha (TNF-α) or IL-1β were found. Inhibition of the mitogen-activated protein kinases JNK1/2, p38, and ERK1/2 modified IL-12p70 production similarly in MIMLh5 and n-MIMLh5, suggesting the induction of the same signaling pathways by the two bacterial preparations. Treatment of DCs with cytochalasin D to inhibit endocytosis before the addition of fluorescently labeled MIMLh5 cells led to a dramatic reduction in the proportion of fluorescencepositive DCs and decreased IL-12 production. Endocytosis and IL-12 production were only marginally affected by cytochalasin D pretreatment when fluorescently labeled n-MIMLh5 was used. Treatment of DCs with fluorescently labeled S-layercoated polystyrene beads (Sl-beads) resulted in much greater uptake of beads, compared to noncoated beads. Prestimulation of DCs with cytochalasin D reduced the uptake of Sl-beads more than plain beads. These findings indicate that the S-layer plays a role in the endocytosis of MIMLh5 by DCs. In conclusion, this study provides evidence that the S-layer of L. helveticus MIMLh5 is involved in endocytosis of the bacterium, which is important for strong Th1-inducing cytokine production.",

keywords = "Cytochalasin D, Cytokines, MAPKs, Nanoparticles, Probiotic",

author = "Valentina Taverniti and Mauro Marengo and Eva Fuglsang and Skovsted, {Helene Marie} and Stefania Arioli and Giacomo Mantegazza and Giorgio Gargari and Stefania Iametti and Francesco Bonomi and Simone Guglielmetti and Hanne Fr{\o}ki{\ae}r",

year = "2019",

doi = "10.1128/aem.00138-19",

language = "English",

volume = "85",

journal = "Applied and Environmental Microbiology",

issn = "0099-2240",

publisher = "American Society for Microbiology",

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T1 - Surface layer of Lactobacillus helveticus MIMLh5 promotes endocytosis by dendritic cells

AU - Taverniti, Valentina

AU - Marengo, Mauro

AU - Fuglsang, Eva

AU - Skovsted, Helene Marie

AU - Arioli, Stefania

AU - Mantegazza, Giacomo

AU - Gargari, Giorgio

AU - Iametti, Stefania

AU - Bonomi, Francesco

AU - Guglielmetti, Simone

AU - Frøkiær, Hanne

PY - 2019

Y1 - 2019

N2 - Surface layers (S-layers) are proteinaceous arrays covering the cell walls of numerous bacteria. Their suggested properties, such as interactions with the host immune system, have been only poorly described. Here, we aimed to elucidate the role of the S-layer from the probiotic bacterial strain Lactobacillus helveticus MIMLh5 in the stimulation of murine bone-marrow-derived dendritic cells (DCs). MIMLh5 induced greater production of interferon beta (IFN-β), interleukin 10 (IL-10), and IL- 12p70, compared to S-layer-depleted MIMLh5 (naked MIMLh5 [n-MIMLh5]), whereas the isolated S-layer was a poor immunostimulator. No differences in the production of tumor necrosis factor alpha (TNF-α) or IL-1β were found. Inhibition of the mitogen-activated protein kinases JNK1/2, p38, and ERK1/2 modified IL-12p70 production similarly in MIMLh5 and n-MIMLh5, suggesting the induction of the same signaling pathways by the two bacterial preparations. Treatment of DCs with cytochalasin D to inhibit endocytosis before the addition of fluorescently labeled MIMLh5 cells led to a dramatic reduction in the proportion of fluorescencepositive DCs and decreased IL-12 production. Endocytosis and IL-12 production were only marginally affected by cytochalasin D pretreatment when fluorescently labeled n-MIMLh5 was used. Treatment of DCs with fluorescently labeled S-layercoated polystyrene beads (Sl-beads) resulted in much greater uptake of beads, compared to noncoated beads. Prestimulation of DCs with cytochalasin D reduced the uptake of Sl-beads more than plain beads. These findings indicate that the S-layer plays a role in the endocytosis of MIMLh5 by DCs. In conclusion, this study provides evidence that the S-layer of L. helveticus MIMLh5 is involved in endocytosis of the bacterium, which is important for strong Th1-inducing cytokine production.

AB - Surface layers (S-layers) are proteinaceous arrays covering the cell walls of numerous bacteria. Their suggested properties, such as interactions with the host immune system, have been only poorly described. Here, we aimed to elucidate the role of the S-layer from the probiotic bacterial strain Lactobacillus helveticus MIMLh5 in the stimulation of murine bone-marrow-derived dendritic cells (DCs). MIMLh5 induced greater production of interferon beta (IFN-β), interleukin 10 (IL-10), and IL- 12p70, compared to S-layer-depleted MIMLh5 (naked MIMLh5 [n-MIMLh5]), whereas the isolated S-layer was a poor immunostimulator. No differences in the production of tumor necrosis factor alpha (TNF-α) or IL-1β were found. Inhibition of the mitogen-activated protein kinases JNK1/2, p38, and ERK1/2 modified IL-12p70 production similarly in MIMLh5 and n-MIMLh5, suggesting the induction of the same signaling pathways by the two bacterial preparations. Treatment of DCs with cytochalasin D to inhibit endocytosis before the addition of fluorescently labeled MIMLh5 cells led to a dramatic reduction in the proportion of fluorescencepositive DCs and decreased IL-12 production. Endocytosis and IL-12 production were only marginally affected by cytochalasin D pretreatment when fluorescently labeled n-MIMLh5 was used. Treatment of DCs with fluorescently labeled S-layercoated polystyrene beads (Sl-beads) resulted in much greater uptake of beads, compared to noncoated beads. Prestimulation of DCs with cytochalasin D reduced the uptake of Sl-beads more than plain beads. These findings indicate that the S-layer plays a role in the endocytosis of MIMLh5 by DCs. In conclusion, this study provides evidence that the S-layer of L. helveticus MIMLh5 is involved in endocytosis of the bacterium, which is important for strong Th1-inducing cytokine production.

KW - Cytochalasin D

KW - Cytokines

KW - MAPKs

KW - Nanoparticles

KW - Probiotic

U2 - 10.1128/aem.00138-19

DO - 10.1128/aem.00138-19

M3 - Journal article

C2 - 30824443

AN - SCOPUS:85065023237

SN - 0099-2240

VL - 85

JO - Applied and Environmental Microbiology

JF - Applied and Environmental Microbiology

IS - 9

M1 - e00138-19

ER -

Surface layer of Lactobacillus helveticus MIMLh5 promotes endocytosis by dendritic cells

Abstract

Keywords

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