Bacteria inducing th1 cells
A technology of bacteria and cells, applied in the field of inducing the proliferation or activation of Th1 cells, kits or non-human animals, disease compositions, inhibiting the proliferation or activation of Th1 cells, proliferation or activation of the composition, can solve complex, The detailed mechanism is not fully clarified, etc.
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Embodiment 1
[0605] The present inventors forcibly orally administered saliva samples from two Crohn's disease (CD) patients to C57BL / 6 (B6) germ-free (GF) mice to produce gnotobiotic mice. Then, the mice of each group were reared in a gnotobiotic isolator for 6 weeks, and immune cells in the small intestine and colonic lamina propria (LP) were investigated.
[0606] The result is as figure 1 and 2 As shown, in the mice administered with the saliva of CD patient #1 (GF+CD #1 mice), no significant changes were observed in intestinal T cells. In contrast, γγ(IFN-g) was confirmed in the lamina propria of the mouse (GF+CD#2 mouse) administered with the saliva of CD patient #2 + CD4 + Significant accumulation of T cells (Th1 cells).
[0607] Therefore, the cluster composition was compared by 16S rRNA sequencing between the saliva microflora of GF mice before administration and the fecal microflora of animals colonized with this microflora (exGF mice).
[0608] The result is as image 3 As...
Embodiment 2
[0633] In order to investigate the mechanism underlying the induction of Th1 cells mediated by Kp-2H7, various studies have been attempted.
[0634] First, it was investigated whether dead bacteria could induce Th1 cells in vivo. That is, Kp-2H7 cultured in vitro was heat-sterilized and orally administered to GF mice via drinking water for 3 weeks. The result is as Figure 18 As shown, pasteurized bacteria affect the frequency of Th1 cells.
[0635] Then, in order to investigate the specific localization of Kp-2H7, the large intestine of mice colonized with Kp-2H7 was investigated by in situ hybridization.
[0636] The result is as Figure 19 As shown, Kp-2H7 was present on the mucin layer, and no evidence of adhesion to or invasion into the epithelial layer was found. These results suggest that Th1 induction is mediated by the activity of viable bacteria localized away from the epithelial layer.
[0637] Next, in order to further elucidate the mechanism, 8 different K.pn...
Embodiment 3
[0651] In order to further clarify the relationship between oral bacteria and Th1 cell induction, two healthy donors (He#1 and He#2) and two patients with active ulcerative colitis (UC) (UC# 1 and UC#2) further saliva samples were obtained orally administered to GF WT B6 mice.
[0652] The result is as Figure 33 As shown, in the large intestine LP of mice (GF+UC#2 mice) inoculated with saliva samples obtained from UC patient #2, significant Th1 cell activity was confirmed to a degree comparable to that of GF+CD#2 mice. accumulate.
[0653] In order to isolate the strain responsible for the detected induction of Th1 cells, the contents of the cecum of GF+UC#2 mice were collected and cultured in vitro. The result is as Figure 34 As shown, the present inventors succeeded in isolating 13 strains showing a profile of the microflora of GF+UC#2 mice.
[0654] In addition, by orally administering a mixture of 13 strains (13-mix) to GF mice, it was possible to completely reproduc...
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