Endosomal recognition of Lactococcus lactis G121 and its RNA by dendritic cells is key to its allergy-protective effects

Karina Stein, Stephanie Brand, André Jenckel, Anna Sigmund, Zhijian James Chen, Carsten J. Kirschning, Marion Kauth, Holger Heine

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Background: Bacterial cowshed isolates are allergy protective in mice; however, the underlying mechanisms are largely unknown. We examined the ability of . Lactococcus lactis G121 to prevent allergic inflammatory reactions. Objective: We sought to identify the ligands and pattern recognition receptors through which . L lactis G121 confers allergy protection. Methods: L lactis G121-induced cytokine release and surface expression of costimulatory molecules by untreated or inhibitor-treated (bafilomycin and cytochalasin D) human monocyte-derived dendritic cells (moDCs), bone marrow-derived mouse dendritic cells (BMDCs), and moDC/naive CD4+ T-cell cocultures were analyzed by using ELISA and flow cytometry. The pathology of ovalbumin-induced acute allergic airway inflammation after adoptive transfer of BMDCs was examined by means of microscopy. Results: L lactis G121-treated murine BMDCs and human moDCs released TH1-polarizing cytokines and induced TH1 T cells. Inhibiting phagocytosis and endosomal acidification in BMDCs or moDCs impaired the release of TH1-polarizing cytokines, costimulatory molecule expression, and T-cell activation on L lactis G121 challenge. In vivo allergy protection mediated by L lactis G121 was dependent on endosomal acidification in dendritic cells (DCs). Toll-like receptor (Tlr) 13 -/- BMDCs showed a weak response to L lactis G121 and were unresponsive to its RNA. The TH1-polarizing activity of L lactis G121-treated human DCs was blocked by TLR8-specific inhibitors, mediated by L lactis G121 RNA, and synergistically enhanced by activation of nucleotide-binding oligomerization domain-containing protein (NOD) 2. Conclusion: Bacterial RNA is the main driver of . L lactis G121-mediated protection against experimentally induced allergy and requires both bacterial uptake by DCs and endosomal acidification. In mice . L lactis G121 RNA signals through TLR13; however, the most likely intracellular receptor in human subjects is TLR8.

Original languageEnglish (US)
JournalJournal of Allergy and Clinical Immunology
DOIs
StateAccepted/In press - Sep 23 2015

Fingerprint

Lactococcus lactis
Dendritic Cells
Hypersensitivity
RNA
Bone Marrow
Monocytes
Cytokines
T-Lymphocytes
Bacterial RNA
Pattern Recognition Receptors
Cytochalasin D
Adoptive Transfer
Toll-Like Receptors
Ovalbumin
Coculture Techniques
Phagocytosis
Microscopy
Flow Cytometry
Nucleotides
Enzyme-Linked Immunosorbent Assay

Keywords

  • Allergy protection
  • Bacterial RNA
  • Dendritic cells
  • Endosomal acidification
  • Lactococcus lactis G121
  • Pattern recognition receptor
  • Toll-like receptor 13
  • Toll-like receptor 8

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Endosomal recognition of Lactococcus lactis G121 and its RNA by dendritic cells is key to its allergy-protective effects. / Stein, Karina; Brand, Stephanie; Jenckel, André; Sigmund, Anna; Chen, Zhijian James; Kirschning, Carsten J.; Kauth, Marion; Heine, Holger.

In: Journal of Allergy and Clinical Immunology, 23.09.2015.

Research output: Contribution to journalArticle

Stein, Karina ; Brand, Stephanie ; Jenckel, André ; Sigmund, Anna ; Chen, Zhijian James ; Kirschning, Carsten J. ; Kauth, Marion ; Heine, Holger. / Endosomal recognition of Lactococcus lactis G121 and its RNA by dendritic cells is key to its allergy-protective effects. In: Journal of Allergy and Clinical Immunology. 2015.
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abstract = "Background: Bacterial cowshed isolates are allergy protective in mice; however, the underlying mechanisms are largely unknown. We examined the ability of . Lactococcus lactis G121 to prevent allergic inflammatory reactions. Objective: We sought to identify the ligands and pattern recognition receptors through which . L lactis G121 confers allergy protection. Methods: L lactis G121-induced cytokine release and surface expression of costimulatory molecules by untreated or inhibitor-treated (bafilomycin and cytochalasin D) human monocyte-derived dendritic cells (moDCs), bone marrow-derived mouse dendritic cells (BMDCs), and moDC/naive CD4+ T-cell cocultures were analyzed by using ELISA and flow cytometry. The pathology of ovalbumin-induced acute allergic airway inflammation after adoptive transfer of BMDCs was examined by means of microscopy. Results: L lactis G121-treated murine BMDCs and human moDCs released TH1-polarizing cytokines and induced TH1 T cells. Inhibiting phagocytosis and endosomal acidification in BMDCs or moDCs impaired the release of TH1-polarizing cytokines, costimulatory molecule expression, and T-cell activation on L lactis G121 challenge. In vivo allergy protection mediated by L lactis G121 was dependent on endosomal acidification in dendritic cells (DCs). Toll-like receptor (Tlr) 13 -/- BMDCs showed a weak response to L lactis G121 and were unresponsive to its RNA. The TH1-polarizing activity of L lactis G121-treated human DCs was blocked by TLR8-specific inhibitors, mediated by L lactis G121 RNA, and synergistically enhanced by activation of nucleotide-binding oligomerization domain-containing protein (NOD) 2. Conclusion: Bacterial RNA is the main driver of . L lactis G121-mediated protection against experimentally induced allergy and requires both bacterial uptake by DCs and endosomal acidification. In mice . L lactis G121 RNA signals through TLR13; however, the most likely intracellular receptor in human subjects is TLR8.",
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AU - Stein, Karina

AU - Brand, Stephanie

AU - Jenckel, André

AU - Sigmund, Anna

AU - Chen, Zhijian James

AU - Kirschning, Carsten J.

AU - Kauth, Marion

AU - Heine, Holger

PY - 2015/9/23

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N2 - Background: Bacterial cowshed isolates are allergy protective in mice; however, the underlying mechanisms are largely unknown. We examined the ability of . Lactococcus lactis G121 to prevent allergic inflammatory reactions. Objective: We sought to identify the ligands and pattern recognition receptors through which . L lactis G121 confers allergy protection. Methods: L lactis G121-induced cytokine release and surface expression of costimulatory molecules by untreated or inhibitor-treated (bafilomycin and cytochalasin D) human monocyte-derived dendritic cells (moDCs), bone marrow-derived mouse dendritic cells (BMDCs), and moDC/naive CD4+ T-cell cocultures were analyzed by using ELISA and flow cytometry. The pathology of ovalbumin-induced acute allergic airway inflammation after adoptive transfer of BMDCs was examined by means of microscopy. Results: L lactis G121-treated murine BMDCs and human moDCs released TH1-polarizing cytokines and induced TH1 T cells. Inhibiting phagocytosis and endosomal acidification in BMDCs or moDCs impaired the release of TH1-polarizing cytokines, costimulatory molecule expression, and T-cell activation on L lactis G121 challenge. In vivo allergy protection mediated by L lactis G121 was dependent on endosomal acidification in dendritic cells (DCs). Toll-like receptor (Tlr) 13 -/- BMDCs showed a weak response to L lactis G121 and were unresponsive to its RNA. The TH1-polarizing activity of L lactis G121-treated human DCs was blocked by TLR8-specific inhibitors, mediated by L lactis G121 RNA, and synergistically enhanced by activation of nucleotide-binding oligomerization domain-containing protein (NOD) 2. Conclusion: Bacterial RNA is the main driver of . L lactis G121-mediated protection against experimentally induced allergy and requires both bacterial uptake by DCs and endosomal acidification. In mice . L lactis G121 RNA signals through TLR13; however, the most likely intracellular receptor in human subjects is TLR8.

AB - Background: Bacterial cowshed isolates are allergy protective in mice; however, the underlying mechanisms are largely unknown. We examined the ability of . Lactococcus lactis G121 to prevent allergic inflammatory reactions. Objective: We sought to identify the ligands and pattern recognition receptors through which . L lactis G121 confers allergy protection. Methods: L lactis G121-induced cytokine release and surface expression of costimulatory molecules by untreated or inhibitor-treated (bafilomycin and cytochalasin D) human monocyte-derived dendritic cells (moDCs), bone marrow-derived mouse dendritic cells (BMDCs), and moDC/naive CD4+ T-cell cocultures were analyzed by using ELISA and flow cytometry. The pathology of ovalbumin-induced acute allergic airway inflammation after adoptive transfer of BMDCs was examined by means of microscopy. Results: L lactis G121-treated murine BMDCs and human moDCs released TH1-polarizing cytokines and induced TH1 T cells. Inhibiting phagocytosis and endosomal acidification in BMDCs or moDCs impaired the release of TH1-polarizing cytokines, costimulatory molecule expression, and T-cell activation on L lactis G121 challenge. In vivo allergy protection mediated by L lactis G121 was dependent on endosomal acidification in dendritic cells (DCs). Toll-like receptor (Tlr) 13 -/- BMDCs showed a weak response to L lactis G121 and were unresponsive to its RNA. The TH1-polarizing activity of L lactis G121-treated human DCs was blocked by TLR8-specific inhibitors, mediated by L lactis G121 RNA, and synergistically enhanced by activation of nucleotide-binding oligomerization domain-containing protein (NOD) 2. Conclusion: Bacterial RNA is the main driver of . L lactis G121-mediated protection against experimentally induced allergy and requires both bacterial uptake by DCs and endosomal acidification. In mice . L lactis G121 RNA signals through TLR13; however, the most likely intracellular receptor in human subjects is TLR8.

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KW - Bacterial RNA

KW - Dendritic cells

KW - Endosomal acidification

KW - Lactococcus lactis G121

KW - Pattern recognition receptor

KW - Toll-like receptor 13

KW - Toll-like receptor 8

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