Method for establishing HBV-CTLs induced hepatitis mouse model
A technology of HBV-CTL and mouse model, which is applied in the field of biomedicine, can solve the problems of easy mutation, infection, and multiple organ damage of the virus, and achieve the effect of reducing the degree of liver tissue damage
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Embodiment 1
[0028] Example 1 HBV-CTLs induced hepatitis mouse model and its construction method
[0029] HBV-CTLs can induce liver tissue damage in HBV transgenic mice:
[0030] After sensitized mice were injected with liver homogenate of HBV transgenic mice, it was observed that the levels of serum transaminases ALT and AST increased transiently, and the spleen volume of sensitized mice increased significantly after repeated stimulation. The isolated HBV-CTLs (preservation number CGMCC No.8753) were intravenously injected into HBV transgenic mice. image 3 The results showed that after the injection of CTLs, the levels of ALT and AST in the serum of HBV transgenic mice were significantly increased; compared with the levels of serum transaminases in mice injected with HBV-CTLs, CTLs derived from the spleen of ordinary BALB / c mice were intravenously injected into HBV In transgenic mice, there was no significant change in serum ALT and AST. After multiple consecutive injections of HBV-CTL...
Embodiment 2
[0032] Example 2 Preparation, isolation and purification of tumor-induced MDSCs
[0033] First, the H22 hepatocellular carcinoma cell line was subcutaneously injected into normal BALB / c mice, 4×10 5 per mouse to establish a liver cancer xenograft tumor model. The size of the tumor was observed the next day. After 2 weeks, when the tumor diameter reached more than 1 cm, the mice were anesthetized and killed, and the femoral bone marrow was collected, and CD11b was sorted using a magnetic sorting system (Stem Cell, Canada) + Myeloid cells, obtained cell marker flow antibody, further purified MDSCs by flow sorting technology, and obtained Gr-1 with a purity of about 95% + CD11b + MDSCs ( Figure 4 , deposit number CGMCC No.8753), for future use.
Embodiment 3
[0034] Example 3 Adoptive reinfusion of MDSCs can reduce liver damage in mice with HBV hepatitis
[0035] MDSCs have antigen-specific and non-specific immunosuppressive effects, and play an important role in inducing immune tolerance. Purify the MDSCs derived from the bone marrow of the tumor-bearing mice isolated in Example 2, and press 5×10 6 MDSCs / only was intravenously reinfused to the hepatitis mice in Example 1, and the control group was injected with the same amount of PBS, and the serum ALT and AST levels of the mice were measured 24 hours later. Figure 5 The results showed that after intravenous reinfusion of MDSCs, the serum ALT and AST levels of HBV transgenic mice were significantly lower than those of mice without MDSCs injection (P<0.05).
[0036] figure 2 The flow chart of establishing HBV-CTLs-induced hepatitis mouse model and MDSCs reinfusion in the present invention.
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