Cellular therapeutic composition for preventing or treating metabolic diseases and comprising Th17 cells as active ingredient
A technology for metabolic diseases and cell therapy, which is applied in the field of cell therapy compositions for the prevention or treatment of metabolic diseases, and can solve the problems of hypoglycemic shock, weight gain and the like
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Embodiment 1
[0059] Embodiment 1. Experimental preparation and experimental method
[0060] 1-1. Preparation of mice
[0061] C57BL / 6 (B6), Foxp3-GFP (B6.Cg-Foxp3 tm2Tc h / J), Rag1-null (B6.129S7-Rag1 tm1Mom / J), B6 / Thy1.1 (B6.PL-Thy1 a / CyJ) and β7 integrin-deficiency (B6.Itgb7 tm1cgn / J) Mice. IL-17A-deficient (C57BL / 6background) mice were provided by Y.C. Sung (Pohang University of Science and Technology, Pohang, Republic of Korea). Mice were placed in the animal room within Pohang University of Science and Technology, and all animal experiments were performed in accordance with protocols approved by the Ethical Review Committee for Animal Experiments at Pohang University of Science and Technology.
[0062] 1-2. Flow cytometry
[0063] All fluorescent-conjugated antibodies used for flow cytometric analysis were purchased from BD Biosciences, eBioscience or Biolegend. After blocking the Fc receptors of cells with anti-CD16 / CD32 (2.4G2, BD) antibody, anti-TCRβ (H57-597), anti-Th...
Embodiment 2
[0091] Example 2: Confirmation of the reduction of small intestinal Th17 cells in obese mice
[0092] In order to confirm the effect of HFD-induced obesity on the intestinal immune system, the following experiments were carried out according to the methods described in Examples 1-2.
[0093] CD4 in the small intestinal mucosal lamina propria (LP) of mice fed normal chow diet (NCD) or high fat diet (high fat diet, HFD) within 10 weeks + The distribution of Th subset (subset) and described each CD4 + The distribution of master transcription factors in the Th subset was confirmed.
[0094] The results are shown in Figure 1, and it was confirmed that small intestinal IFN-γ in HFD mice compared with NCD mice + Increased Th1 cells, IL-17A + The proportion of Th17 cells was greatly reduced ( Figure 1a and 1b ), and ob / ob mice, a genetic model of obesity and type 2 diabetes, also had a decrease in Th17 cells. In addition, it was confirmed that RORγ in the small intestine of HF...
Embodiment 3
[0096] Example 3: Confirmation that vitamin A deficiency in obese mice results in a reduction in small intestinal Th17 cells
[0097] In order to confirm whether further reduction of Th17 cells due to vitamin A-deficiency, which is closely related to the exacerbation of metabolic disorders, would exacerbate HFD-induced obesity, experiments were conducted as described in Examples 1-3.
[0098] As a result, as shown in Figure 2, it was confirmed that SI Th17 cells were reduced in mice consuming a vitamin A-deficient high-fat diet (VAD) compared to mice consuming a vitamin A-enriched high-fat diet (VAS) . The proportion of Th1 cells was slightly increased. In contrast, Treg cells were not affected by VAD ( Figure 2a ). In addition, the total mass of VAD mice was much higher than that of VAS mice ( Figure 2b ), and the fat mass of epididymis in VAD mice is also higher ( Figure 2c ). In addition, it was confirmed that the fasting glucose value and insulin value, glucose ...
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