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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

Inactive Publication Date: 2014-12-24
PEOPLES HOSPITAL PEKING UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Concanavalin can induce abnormal activation of T cells and cause liver tissue damage, but the liver injury induced by concanavalin is antigen-nonspecific and dose-dependent, mainly due to the participation of activated CD4+ T cells, and can cause visceral damage. organ damage
Some other studies try to dynamically inject animals with adenovirus and retrovirus recombinant vectors carrying viral genes to establish a hepatitis model of viral infection. The disadvantage is that the virus is prone to mutation and is affected by the virus and the host.

Method used

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  • Method for establishing HBV-CTLs induced hepatitis mouse model
  • Method for establishing HBV-CTLs induced hepatitis mouse model
  • Method for establishing HBV-CTLs induced hepatitis mouse model

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention provides a method for establishing an HBV-CTLs induced hepatitis mouse model. The HBV-CTLs induced hepatitis mouse model is established by injecting an HBV antigen-specificity cytotoxic T cell HBV-CTL with the preservation number of CGMCC No. 8753 into an HBV transgenic mouse in an intravenous injection manner. The research shows that the hepatitis mouse model established by using the method can be used for overcoming the HBV virus species specificity, the HBV-CTLs can simulate liver tissue damage mediated after hepatitis virus infection in the physiological state, and the liver tissue damage degree can be reduced by outputting back tumor induced MDSCs. On the basis that a hepatitis mouse animal model is established, novel strategies for treating hepatitis is proposed, and novel ideas are provided for hepatitis treatment.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a method for constructing an HBV-CTLs-induced hepatitis mouse model. Background technique [0002] Chronic hepatitis (chronic hepatitis) is one of the most common diseases that threaten human health. Various factors, such as chronic viral infection, excessive alcohol intake, metabolic diseases and immune disorders, can lead to long-term chronic liver tissue damage, resulting in liver tissue damage. Lesions, and even acute-on-chronic hepatitis and primary liver cancer. In countries such as Europe and the United States, chronic hepatitis is mainly caused by excessive alcohol intake. For example, more than 60% of patients with liver cirrhosis and liver cancer have a history of alcoholic liver disease. In China, chronic viral infection is the main cause of chronic hepatitis, liver cirrhosis, and liver cancer, mainly including hepatitis B virus (HBV) and hepatitis C virus (HCV) infection. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/89C12N5/0783C12N5/071A61K35/56A61P31/20
Inventor 张恒辉陈红松贺改霞张明徽陈栋伟
Owner PEOPLES HOSPITAL PEKING UNIV