Method for constructing chronic transfection model of hepatitis B virus

Abstract

The invention belongs to the field of biomedicine, and particularly relates to a method for constructing a chronic transfection model of a hepatitis B virus. The method disclosed by the invention comprises the following steps: transfecting HBV (hepatitis B virus) plasmids into a model-preparing experimental mouse (preferably adopting male C3H / HeN of 4-8-week old), and constructing the chronic transfection model of the HBV. The chronic transfection model of the hepatitis B virus constructed by the method can be used for researching specific action mechanism of single or multiple genes in an HBV pathogenic process; transgenic modification of the mouse is not needed; the method is free of limitations such as immunologic tolerance, easy to obtain, and low in cost; the problem of long-term shortage of an animal model suitable for the HBV in related technical fields can be solved; and a good animal model is provided for further research on life history and chronic mechanism of the HBV and screening of vaccines and new medicines.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a chronic transfection model of hepatitis B virus, in particular to a method for constructing a chronic transfection model of hepatitis B virus. Background technique [0002] As far as hepatitis B virus (hepatitis B virus, HBV) is concerned, there are about 2 billion infected people worldwide, of which 350 million people become chronic hepatitis B virus carriers, and there are about 90 million hepatitis B virus carriers in China. According to reports, chronic infection of hepatitis B virus often leads to serious diseases such as liver cirrhosis and liver cancer; the direct medical expenses for hepatitis and liver disease in China are as high as more than 100 billion yuan every year. The current treatment measures for the disease include nucleoside drugs such as lamivudine, adefovir dipivoxil, etc., but the drugs can only inhibit virus replication to a certain extent and easily cause virus...

AI Technical Summary

Problems solved by technology

The current treatment measures for the disease include nucleoside drugs such as lamivudine, adefovir dipivoxil, etc., but the drugs can only inhibit virus replication to a certain extent and easily cause virus drug-resistant mutations, while conventional interferon -α (IFN-α) and pegylated IFN-α are only effective in some patients (less than 30% response rate)

[0004] The prior art has been published, and the animal infection model established for HBV research, however, the existing HBV infection animal models have great limitations

For example, chimpanzees are endangered animals and are expensive and involve ethical issues, so their use is strictly restricted; in fact, experimental research on non-human primates has been banned in Europe, and the NIH in the United States does not support it; although tree shrews can infect HBV, but cannot continue to replicate to cause chronic infection, there is a certain gap with clinical practice; the infection model established by other hepadophilic viruses such as duck hepatitis B virus also has different gene transcription regulation mechanisms, and the immune system of poultry and humans is far different; Commonly used HBV mouse models, such as transgenic mouse models, HBV-infected human liver chimeric mouse models, and acute / chronic HBV transfected mouse models, have their own characteristics, which have promoted HBV-related research to a certain extent. However, These mouse models still have problems such as autoimmune tolerance, lack of immune system or low level of HBV replication and low modeling rate in terms of immune pathogenesis, vaccine evaluation and drug screening.

[0005] Some studies used the high-pressure hydrodynamic method to transfect the pT-MCS-HBV1.3 plasmid to establish a HBV acute transfection mouse model in B10.D2 mice, but the duration was short (less than 15 days); Huang LR et al. (PNAS, 2006;103:17862-7) established a chronic infection model in C57BL / 6 mice by using high-pressure hydrodynamic method to transfer pAAV / HBV1.2, the virus antigen in the blood can be maintained for more than 6 months, but the chronicity rate is low (< 40%), while in BALB / c only acute models can be constructed (viral antigens cleared within 1 month); Chang W et al. (World J Gastroenterol, 2005; 11:6631-7) used high pressure hydrodynamic transfection pHBV3.6 plasmid to C3H / HeN mice of 8-12 weeks, but only an acute transfection model can be constructed (viral antigen can only be maintained for one week);

[0006] To sum up, the HBV transfection chronicity modulus model constructed based on the high-pressure hydrodynamic method in the prior art still has problems to be overcome and needs to be further optimized

Images