Department of Gastroenterology, Juntendo University School of Medicine, Tokyo 113-8421, Japan.
World J Gastroenterol. 2011 Sep 21;17(35):4017-22. doi: 10.3748/wjg.v17.i35.4017.
To study the production and secretion of corticotropin-releasing factor (CRF) by dendritic cells and the influence of commensal bacteria.
JAWSII cells (ATCC CRL-11904), a mouse dendritic cell line, were seeded into 24-well culture plates and grown for 3 d. Commensal bacterial strains of Clostridium clostrodiiforme (JCM1291), Bacteroides vulgatus (B. vulgatus) (JCM5856), Escherichia coli (JCM1649), or Fusobacterium varium (F. varium) (ATCC8501) were added to the cells except for the control well, and incubated for 2 h. After incubation, we performed enzyme-linked immunosorbent assay for the cultured medium and reverse transcription polymerase chain reaction for the dendritic cells, and compared these values with controls.
The level of CRF secretion by control dendritic cells was 40.4 ± 6.2 pg/mL. The CRF levels for cells incubated with F. varium and B. vulgatus were significantly higher than that of the control (P < 0.0001). CRF mRNA was present in the control sample without bacteria, and CRF mRNA levels in all samples treated with bacteria were above that of the control sample. F. varium caused the greatest increase in CRF mRNA expression.
Our results suggest that dendritic cells produce CRF, a process augmented by commensal bacteria.
研究树突状细胞(DC)产生和分泌促肾上腺皮质释放因子(CRF)的情况及其受共生菌的影响。
JAWSII 细胞(ATCC CRL-11904)是一种小鼠树突状细胞系,将其接种到 24 孔培养板中培养 3 天。除对照孔外,将共生菌 Clostridium clostrodiiforme(JCM1291)、Bacteroides vulgatus(B. vulgatus)(JCM5856)、Escherichia coli(JCM1649)或 Fusobacterium varium(F. varium)(ATCC8501)添加到细胞中,并孵育 2 小时。孵育后,我们对培养物进行酶联免疫吸附测定,对树突状细胞进行逆转录聚合酶链反应,并与对照进行比较。
对照树突状细胞分泌的 CRF 水平为 40.4±6.2 pg/mL。与对照相比,与 F. varium 和 B. vulgatus 孵育的细胞中 CRF 水平显著升高(P<0.0001)。对照样本中存在无细菌的 CRF mRNA,所有用细菌处理的样本中的 CRF mRNA 水平均高于对照样本。F. varium 引起 CRF mRNA 表达的最大增加。
我们的结果表明,树突状细胞产生 CRF,共生菌可增强这一过程。