New method for treating type II diabetes through immunity adjustment and control
A modulatory, compound-based technology for biomedical applications that addresses the lack of proof-of-concept studies
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Embodiment 1
[0207] Example 1: Effect of long-term insulin injection on mouse insulin sensitivity
[0208] In this example, wild-type mice received daily intraperitoneal injection of PBS or insulin (0.5 U / Kg) for two months.
[0209] The result is as figure 1 Insulin injection in vivo can induce a large amount of CD73 low Adipose regulatory T cells. CD73 expression was greatly suppressed in insulin-injected mouse adipose regulatory T cells, but CD39 levels remained largely unchanged (Fig. 1a–c).
[0210] Further, glucose tolerance test (GTT) and insulin tolerance test (ITT) were performed on mice to study CD73 low Relationship between fat regulatory T cells and insulin sensitivity. The results showed that insulin-treated mice had greater blood glucose fluctuations and impaired insulin sensitivity (Fig. 1d). Thus, these data suggest that chronic insulin injection induces the accumulation of CD73 low Adipose regulatory T cells and cause insulin resistance.
Embodiment 2
[0211] Example 2: CD73 low and CD73 hi A Comparison of the Suppressive Ability of Adipose Regulatory T Cells
[0212] CD4 sorting from wild-type mice + CD25-YFP-Teff cells labeled with CellTrace Violet dye. and compared it with CD4 sorted from wild-type mice + CD25 hi YFP + CD73 hi Adipose regulatory T cells or CD4 + CD25 hi YFP + CD73 low Adipose regulatory T cell co-culture. The cell mixture system was stimulated with anti-CD3 / CD28 antibody-coupled magnetic beads, and the proliferation of responding cells was measured three days later. Afterwards, the proportion of responding Teff cells proliferating was further measured.
[0213] The result is as figure 2 As shown in a-d, in vitro tests suggest that CD73 hi Adiporegulatory T cells were more suppressive (Fig. 2a,b), indicating their protective effect on adipose inflammatory responses. In addition, CD73 low Fat-regulated production of large amounts of the inflammatory factor IFN-γ exhibited a phenotype resemb...
Embodiment 3
[0217] Example 3: Adenosine or Adenosine Receptor A 2A Effects of receptor agonists on cells
[0218] In this example, adenosine or adenosine receptor A 2A Th1 cells and undifferentiated M0 macrophages were directly treated with receptor agonists to test the direct effect of these two small molecules on the cells.
[0219] The result is as image 3 Indicated, adenosine (10 μM) or adenosine receptor A 2A After Th1 cells were directly treated with receptor agonist (1 μM), the cells secreted less inflammatory cytokines IL-2 and IFN-γ ( image 3 a-c), suggesting that this inhibitory effect acts directly on Th1 cells.
[0220] And, adenosine or adenosine receptor A 2A After receptor agonists directly treat M0 macrophages, the cells up-regulate the marker molecule CD206 of M2 macrophages ( image 3 d and e), while macrophages secrete less pro-inflammatory cytokine IL-1β ( image 3 f), secrete more immunosuppressive cytokine IL-10 ( image 3 g).
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