Journal
FRONTIERS IN IMMUNOLOGY
Volume 7, Issue -, Pages -Publisher
FRONTIERS MEDIA SA
DOI: 10.3389/fimmu.2016.00601
Keywords
probiotic; Ca2+ signaling; intestinal epithelial cell; intravital imaging; Lactococcus; Bacillus subtilis; small intestine; germ-free mouse
Categories
Funding
- Ministry of Education, Culture, Sports, Science and Technology of Japan
- All Japan Coffee Association
- Mishima Kaiun Memoria
- Yamada Foundation
- Joint Usage/Research Program of Medical Research Institute, Tokyo Medical and Dental University
- Strategic International Collaborative Research Program (SICORP)
- Japan Society for the Promotion Science (JSPS) [15H04504]
- Cross-ministerial Strategic Innovation Promotion Program (SIP)
- Japan Agency for Medical Research and Development, AMED
- Yakult Bio-Science Foundation
- Takano Bio-Science Foundation
- Canon Foundation
- Futaba Electronics Memorial Foundation
- Grants-in-Aid for Scientific Research [15H04504] Funding Source: KAKEN
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Probiotics, such as lactic acid bacteria (LAB) and Bacillus subtilis var. natto, have been shown to modulate immune responses. It is important to understand how probiotic bacteria impact intestinal epithelial cells (IECs), because IECs are the first line of defense at the mucosal surface barrier and their activities substantially affect the gut microenvironment and immunity. However, to date, their precise mechanism remains unknown due to a lack of analytical systems available for live animal models. Recently, we generated a conditional Ca2+ biosensor Yellow Cameleon (YC3.60) transgenic mouse line and established 5D (x, y, z, time, and Ca2+) intravital imaging systems of lymphoid tissues including those in Peyer's patches and bone marrow. In the present study, we further advance our intravital imaging system for intestinal tracts to visualize IEC responses against orally administrated food compounds in real time. Using this system, heat-killed B. subtilis natto, a probiotic TTCC012 strain, is shown to directly induce Ca2+ signaling in IECs in mice housed under specific pathogen-free conditions. In contrast, this activation is not observed in the Lactococcus lactis strain C60; however, when we generate germ-free YC3.60 mice and observe the LAB stimulation of IECs in the absence of gut microbiota, C60 is capable of inducing Ca2+ signaling. This is the first study to successfully visualize the direct effect of probiotics on IECs in live animals. These data strongly suggest that probiotic strains stimulate IECs under physiological conditions and that their activity is affected by the microenvironment of the small intestine, such as commensal bacteria.
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