Cargando…

Phosphoinositide 3-Kinaseγ Controls the Intracellular Localization of CpG to Limit DNA-PKcs-Dependent IL-10 Production in Macrophages

Synthetic oligodeoxynucleotides containing unmethylated CpG motifs (CpG) stimulate innate immune responses. Phosphoinositide 3-kinase (PI3K) has been implicated in CpG-induced immune activation; however, its precise role has not yet been clarified. CpG-induced production of IL-10 was dramatically in...

Descripción completa

Detalles Bibliográficos
Autores principales: Hazeki, Kaoru, Kametani, Yukiko, Murakami, Hiroki, Uehara, Masami, Ishikawa, Yuki, Nigorikawa, Kiyomi, Takasuga, Shunsuke, Sasaki, Takehiko, Seya, Tsukasa, Matsumoto, Misako, Hazeki, Osamu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3203906/
https://www.ncbi.nlm.nih.gov/pubmed/22053215
http://dx.doi.org/10.1371/journal.pone.0026836
_version_ 1782215151540240384
author Hazeki, Kaoru
Kametani, Yukiko
Murakami, Hiroki
Uehara, Masami
Ishikawa, Yuki
Nigorikawa, Kiyomi
Takasuga, Shunsuke
Sasaki, Takehiko
Seya, Tsukasa
Matsumoto, Misako
Hazeki, Osamu
author_facet Hazeki, Kaoru
Kametani, Yukiko
Murakami, Hiroki
Uehara, Masami
Ishikawa, Yuki
Nigorikawa, Kiyomi
Takasuga, Shunsuke
Sasaki, Takehiko
Seya, Tsukasa
Matsumoto, Misako
Hazeki, Osamu
author_sort Hazeki, Kaoru
collection PubMed
description Synthetic oligodeoxynucleotides containing unmethylated CpG motifs (CpG) stimulate innate immune responses. Phosphoinositide 3-kinase (PI3K) has been implicated in CpG-induced immune activation; however, its precise role has not yet been clarified. CpG-induced production of IL-10 was dramatically increased in macrophages deficient in PI3Kγ (p110γ(−/−)). By contrast, LPS-induced production of IL-10 was unchanged in the cells. CpG-induced, but not LPS-induced, IL-10 production was almost completely abolished in SCID mice having mutations in DNA-dependent protein kinase catalytic subunit (DNA-PKcs). Furthermore, wortmannin, an inhibitor of DNA-PKcs, completely inhibited CpG-induced IL-10 production, both in wild type and p110γ(−/−) cells. Microscopic analyses revealed that CpG preferentially localized with DNA-PKcs in p110γ(−/−) cells than in wild type cells. In addition, CpG was preferentially co-localized with the acidic lysosomal marker, LysoTracker, in p110γ(−/−) cells, and with an early endosome marker, EEA1, in wild type cells. Over-expression of p110γ in Cos7 cells resulted in decreased acidification of CpG containing endosome. A similar effect was reproduced using kinase-dead mutants, but not with a ras-binding site mutant, of p110γ. Thus, it is likely that p110γ, in a manner independent of its kinase activity, inhibits the acidification of CpG-containing endosomes. It is considered that increased acidification of CpG-containing endosomes in p110γ(−/−) cells enforces endosomal escape of CpG, which results in increased association of CpG with DNA-PKcs to up-regulate IL-10 production in macrophages.
format Online
Article
Text
id pubmed-3203906
institution National Center for Biotechnology Information
language English
publishDate 2011
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-32039062011-11-03 Phosphoinositide 3-Kinaseγ Controls the Intracellular Localization of CpG to Limit DNA-PKcs-Dependent IL-10 Production in Macrophages Hazeki, Kaoru Kametani, Yukiko Murakami, Hiroki Uehara, Masami Ishikawa, Yuki Nigorikawa, Kiyomi Takasuga, Shunsuke Sasaki, Takehiko Seya, Tsukasa Matsumoto, Misako Hazeki, Osamu PLoS One Research Article Synthetic oligodeoxynucleotides containing unmethylated CpG motifs (CpG) stimulate innate immune responses. Phosphoinositide 3-kinase (PI3K) has been implicated in CpG-induced immune activation; however, its precise role has not yet been clarified. CpG-induced production of IL-10 was dramatically increased in macrophages deficient in PI3Kγ (p110γ(−/−)). By contrast, LPS-induced production of IL-10 was unchanged in the cells. CpG-induced, but not LPS-induced, IL-10 production was almost completely abolished in SCID mice having mutations in DNA-dependent protein kinase catalytic subunit (DNA-PKcs). Furthermore, wortmannin, an inhibitor of DNA-PKcs, completely inhibited CpG-induced IL-10 production, both in wild type and p110γ(−/−) cells. Microscopic analyses revealed that CpG preferentially localized with DNA-PKcs in p110γ(−/−) cells than in wild type cells. In addition, CpG was preferentially co-localized with the acidic lysosomal marker, LysoTracker, in p110γ(−/−) cells, and with an early endosome marker, EEA1, in wild type cells. Over-expression of p110γ in Cos7 cells resulted in decreased acidification of CpG containing endosome. A similar effect was reproduced using kinase-dead mutants, but not with a ras-binding site mutant, of p110γ. Thus, it is likely that p110γ, in a manner independent of its kinase activity, inhibits the acidification of CpG-containing endosomes. It is considered that increased acidification of CpG-containing endosomes in p110γ(−/−) cells enforces endosomal escape of CpG, which results in increased association of CpG with DNA-PKcs to up-regulate IL-10 production in macrophages. Public Library of Science 2011-10-28 /pmc/articles/PMC3203906/ /pubmed/22053215 http://dx.doi.org/10.1371/journal.pone.0026836 Text en Hazeki et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Hazeki, Kaoru
Kametani, Yukiko
Murakami, Hiroki
Uehara, Masami
Ishikawa, Yuki
Nigorikawa, Kiyomi
Takasuga, Shunsuke
Sasaki, Takehiko
Seya, Tsukasa
Matsumoto, Misako
Hazeki, Osamu
Phosphoinositide 3-Kinaseγ Controls the Intracellular Localization of CpG to Limit DNA-PKcs-Dependent IL-10 Production in Macrophages
title Phosphoinositide 3-Kinaseγ Controls the Intracellular Localization of CpG to Limit DNA-PKcs-Dependent IL-10 Production in Macrophages
title_full Phosphoinositide 3-Kinaseγ Controls the Intracellular Localization of CpG to Limit DNA-PKcs-Dependent IL-10 Production in Macrophages
title_fullStr Phosphoinositide 3-Kinaseγ Controls the Intracellular Localization of CpG to Limit DNA-PKcs-Dependent IL-10 Production in Macrophages
title_full_unstemmed Phosphoinositide 3-Kinaseγ Controls the Intracellular Localization of CpG to Limit DNA-PKcs-Dependent IL-10 Production in Macrophages
title_short Phosphoinositide 3-Kinaseγ Controls the Intracellular Localization of CpG to Limit DNA-PKcs-Dependent IL-10 Production in Macrophages
title_sort phosphoinositide 3-kinaseγ controls the intracellular localization of cpg to limit dna-pkcs-dependent il-10 production in macrophages
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3203906/
https://www.ncbi.nlm.nih.gov/pubmed/22053215
http://dx.doi.org/10.1371/journal.pone.0026836
work_keys_str_mv AT hazekikaoru phosphoinositide3kinasegcontrolstheintracellularlocalizationofcpgtolimitdnapkcsdependentil10productioninmacrophages
AT kametaniyukiko phosphoinositide3kinasegcontrolstheintracellularlocalizationofcpgtolimitdnapkcsdependentil10productioninmacrophages
AT murakamihiroki phosphoinositide3kinasegcontrolstheintracellularlocalizationofcpgtolimitdnapkcsdependentil10productioninmacrophages
AT ueharamasami phosphoinositide3kinasegcontrolstheintracellularlocalizationofcpgtolimitdnapkcsdependentil10productioninmacrophages
AT ishikawayuki phosphoinositide3kinasegcontrolstheintracellularlocalizationofcpgtolimitdnapkcsdependentil10productioninmacrophages
AT nigorikawakiyomi phosphoinositide3kinasegcontrolstheintracellularlocalizationofcpgtolimitdnapkcsdependentil10productioninmacrophages
AT takasugashunsuke phosphoinositide3kinasegcontrolstheintracellularlocalizationofcpgtolimitdnapkcsdependentil10productioninmacrophages
AT sasakitakehiko phosphoinositide3kinasegcontrolstheintracellularlocalizationofcpgtolimitdnapkcsdependentil10productioninmacrophages
AT seyatsukasa phosphoinositide3kinasegcontrolstheintracellularlocalizationofcpgtolimitdnapkcsdependentil10productioninmacrophages
AT matsumotomisako phosphoinositide3kinasegcontrolstheintracellularlocalizationofcpgtolimitdnapkcsdependentil10productioninmacrophages
AT hazekiosamu phosphoinositide3kinasegcontrolstheintracellularlocalizationofcpgtolimitdnapkcsdependentil10productioninmacrophages