Human-derived bacteria that induce proliferation or accumulation of regulatory t cells
A regulatory and bacterial technology, applied in the direction of medical raw materials derived from bacteria, bacteria, animal cells, etc.
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Embodiment 1
[0238] Example 1: First, it was investigated whether the accumulation of regulatory T cells (Treg cells) in the lamina propria of the colon was dependent on commensal bacteria. Specifically, lymphocytes were isolated from the peripheral lymph nodes (pLN) of Balb / c mice fed in the absence of specific pathogenic bacteria (SPF) or from the colon or small intestine (SI) lamina propria of the mice. CD4 and Foxp3 were stained by antibodies. Then, CD4 was analyzed by flow cytometry + Foxp3 in lymphocytes + cell ratio. The results showed that Foxp3 + Treg cells are present at high frequency in the lamina propria of the gastrointestinal tract of mice maintained in a specific pathogen free (SPF) environment, especially the lamina propria of the colon. In addition, it was also found that Foxp3 in the colon lamina propria + The number of Treg cells gradually increased until three months after birth, while Foxp3 in peripheral lymph nodes + The number of Treg cells remains essential...
Embodiment 2
[0240]Example 2: Next, it was investigated whether the temporal accumulation of Treg cells in the colon found in Example 1 is related to the colonization of the intestinal commensal microbiota. Specifically, the small intestine, colon, and peripheral lymph nodes isolated from mice (8 weeks old: Balb / c mice, IQI mice, and C57BL / 6 mice) fed in a germ-free (GF) or SPF environment were analyzed. CD4 expression and Foxp3 expression in lymphocytes. Similar results were obtained in three or more independent experiments.
[0241] In addition, lamina propria lymphocytes were collected from SPF mice and GF mice (Balb / c mice or C57BL / 6 mice). CD4 and Foxp3 were stained with antibodies. Then, lamina propria lymphocytes were analyzed by FACS.
[0242] In addition, lymphocytes were isolated from colonic lamina propria, small intestinal lamina propria (SI), Peyer's patches (PP) and mesenteric lymph nodes (MLN) of mice administered antibiotics orally with water for eight weeks (SPF C57BL / 6...
Embodiment 3
[0251] Example 3: Next, it was directly examined whether the decrease in the number of Treg cells in the colonic lamina propria of GF mice shown in Example 2 was due to the absence of microbiota. Specifically, a feces suspension (customized) of B6SPF mice purchased from The Jackson Laboratory was orally administered to GF-IQI mice. Three weeks after administration, lymphocytes were isolated from the colonic lamina propria and analyzed for CD4 + Expression of Foxp3 in lymphocytes. The results showed that the number of Treg cells in the lamina propria of the small intestine was unchanged. However, the number of Treg cells in the colonic lamina propria was significantly increased. Thus, indicating that the host-microbe interaction in the colonic lamina propria Foxp3 + Treg cells play an important role in the accumulation of Treg cells, while the accumulation of Treg cells in the lamina propria of the small intestine has a different mechanism.
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