Phosphoinositide 3-kinaseγ controls the intracellular localization of CpG to limit DNA-PKcs-dependent IL-10 production in macrophages.
Phosphoinositide 3-kinaseγ controls the intracellular localization of CpG to limit DNA-PKcs-dependent IL-10 production in macrophages.
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DOI：
10.1371/journal.pone.0026836
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Hazeki O
中科院分区：
文献类型：
--
作者：
Hazeki K;Kametani Y;Murakami H;Uehara M;Ishikawa Y;Nigorikawa K;Takasuga S;Sasaki T;Seya T;Matsumoto M;Hazeki O
含有未甲基化CpG基序（CpG）的合成寡脱氧核苷酸刺激先天性免疫应答。磷酸肌醇3-激酶（PI 3 K）与CpG诱导的免疫激活有关;然而，其确切作用尚未阐明。CpG诱导的IL-10产生在PI 3 K γ（p110γ−/−）缺陷的巨噬细胞中显著增加。相比之下，LPS诱导的IL-10的产生在细胞中没有变化。CpG诱导的，但不是LPS诱导的，IL-10的生产几乎完全取消在SCID小鼠的DNA依赖性蛋白激酶催化亚基（DNA-PKcs）突变。此外，在野生型和p110γ−/−细胞中，DNA-PKcs抑制剂渥曼青霉素完全抑制CpG诱导的IL-10产生。显微镜分析显示，CpG优先定位于p110γ−/−细胞中的DNA-PKcs，而不是野生型细胞。此外，CpG在p110γ−/−细胞中优先与酸性溶酶体标记物LysoTracker共定位，在野生型细胞中优先与早期内体标记物EEA 1共定位。p110γ在Cos 7细胞中的过表达导致含CpG的内体的酸化减少。使用p110γ的激酶死亡突变体可以重现类似的效果，但使用ras结合位点突变体则不行。因此，p110γ可能以不依赖于其激酶活性的方式抑制含CpG的内体的酸化。人们认为p110γ−/−细胞中含CpG的内体酸化增加会强制CpG的内体逃逸，这导致CpG与DNA-PKcs的结合增加，从而上调巨噬细胞中IL-10的产生。
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.
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影响因子：
30.5
作者：
Fukao, T;Tanabe, M;Koyasu, S
通讯作者：
Koyasu, S
影响因子：
82.9
作者：
Barber, DF;Bartolomé, A;Carrera, AC
通讯作者：
Carrera, AC
影响因子：
16.8
作者：
Fukao, T;Koyasu, S
通讯作者：
Koyasu, S
影响因子：
30.5
作者：
通讯作者：
--
DOI：
10.1073/pnas.0702663104
发表时间：
2007-05-08
影响因子：
11.1
作者：
Chang, James D.;Sukhova, Galina K.;Cantley, Lewis C.
通讯作者：
Cantley, Lewis C.