Chronic hepatitis B (CHB) caused by hepatitis B virus (HBV) infection and associated liver cirrhosis (L.C). Hepatocellular carcinoma (HCC) and liver failure are important medical and public health problems worldwide. Hepatitis B virus is species-specific and tissue-specific, and can be excreted through various body fluids, such as blood, semen, vaginal secretions, saliva, breast milk, menstruation, tears, urine and sweat. Currently, animal models of hepatitis Bvirus infection include non-human primates, transgenic mice, transfected mice, and immunodeficient chimeric liver mice.
Principle
Based on classical genetics, molecular genetics, structural genetics and DNA recombination technology, the HBV genome was introduced into the genome of the animal chromosome to stabilize the integration of exogenous genes in the genome of the animal, and can be inherited to the offspring of the animal.
The target gene is rapidly injected from the tail vein of mice with a large dose of the transfected gene to obtain a high level of expression of the target gene in the liver of the mice, and using the adeno-associated virus, which has a high degree of hepatophilicity, the HBV genome is transduced into the liver tissue of the mice, and the HBV is expressed in the body of the mice.
The human liver chimeric mouse model involves the transplantation of human hepatocytes into immunodeficient mice, where normal human hepatocytes can grow continuously in the mouse and can therefore be infected with human HBV.
Non-human primates and humans have similarities in physiological and biochemical functions and genomes.
DHBV and HBV belong to the same family of hepatophilic DNA viruses, with 40% nucleotide sequence homology. The susceptibility to DHBV is related to the autoimmunity of ducks. 1~3 days old ducklings with imperfect immune system are easy to be infected by DHBV, and they can produce high level of chronic virus carrying state.
Appliance
The HBV whole genome transgenic animals can be used to study the replication process of HBV at the molecular level. Preclinical evaluation of antiviral drugs, study of drug efficacy, pharmacokinetics and toxicology, also can be used to evaluate the efficacy of immunotherapy.HBV whole genome transgenic animals can be used to study the mechanism of viral clearance and immune damage during HBV infection.HBV specific genome transgenic animals can be used for the study of the function of HBV-specific genes, the role of natural immunity in the immunomodulation of HBV fever, and the role of drugs and vaccines. The HBV-specific genomic transgenic animals can be used for HBV-specific gene function studies, HBV fever immunomodulation and natural immunity, drug and vaccine evaluation, HBV infection mechanism studies, antiviral drug pharmacodynamics and therapeutic vaccine evaluation. It cannot be used to study the interaction between the organism and the virus, especially the mechanism of immune clearance. It can be used to study the mechanism of HBV infection and the mechanism of immune clearance of the virus, and it can be used as a model for the evaluation of the pharmacodynamics of new anti-HBV drugs. It is suitable for research on the biological characteristics of HBV, pathogenic mechanism and evaluation of anti-HBV drugs.
Operation method
Non-human primate models
Principle
Non-human primates share similarities in physiology, biochemical functions and genomes with humans.
Materials and Instruments
Material: chimpanzee, gibbon, rhesus monkey male, bear monkey, tree quail Move The basic procedure in the nonhuman primate model can be divided into the following steps: . Quercus sabdariffa: Sera from HBV-infected patients with HBeAg-positive, HBsAg-positive, HBcAb-positive, and with HBV DNA copies of more than 107 copies/ml should be used The serum from HBV-infected patients with HBV DNA copy number of 107 copies/ml or more was used as the source of infection. 300 ul of serum containing 3 x 106 HBV DNA copies/ml was injected intramuscularly or intraperitoneally into 1-30 days old juvenile tree thrips; 500 ul of serum containing 5 x 106 HBV DNA copies/ml was injected into the femoral vein of 31 days old to 4 months old tree thrips. After 3 days, 500 ul of the same serum was injected intraperitoneally. B. Cyclophosphamide (150 mg/kg) was injected intraperitoneally on days 4 and 3 before bacterial inoculation. Caveat 1. The chimpanzee is currently the only animal model of natural HBV infection. However, it is expensive and difficult to promote. 2. the long arm microtome produces significant serodiscretionary and hepatic histologic changes. 3. the rhesus monkey is selective for HBV-infected material. 3. rhesus monkeys are selective about the HBV material they are infected with. After infection, HBsAg levels are low, and there are no biochemical or histological liver lesions. 4. in rhesus monkeys, HBsAg is detectable in the liver, but the signal is weak, and HBV DNA titers in hepatocytes and serum are low or undetectable. 5. in tree HBV-infected trees, HBsAg is detectable in the liver, but the signal is weak. 5. Tree HBV infection with arboreal enzymes is an acute self-limiting infection. 90% of acute infections, there is expression of HBV DNA in liver tissues, HBsAg appears in the blood, followed by HBsAb, HBeAb, HBcAb, and the virus is cleared very quickly. 56% of the time, the chronic infection can last more than 1 year, and 33% of the time it can last more than 2 years, and the rate of HBsAg positivity in the hepatocytes is 80%. 80%. For more product details, please visit Aladdin Scientific website.
A. Chimpanzees: chimpanzees are injected intravenously with 1 ml of serum containing hepatitis B virus ayw 1undefined or adw 10 or adr ≥ 1undefined or ayr ≥ 10° infectious agent i CID); intravenous injection of saliva and semen containing HBsAg-positive human saliva and semen ( Alter); subcutaneous Inoculation with cloned HBV DNA (Will H. 1982); Hepatitis B virus human plasma dropped onto the surface of chimpanzee corneas; animals developed HBV infection 9 weeks later (Band, 1982). Semen from hepatitis B surface antigen-positive patients was injected into the vagina of chimpanzees.
B. Gibbon: HBsAg(+), HBeAg(+) carrier semen, titer 1:1000 or greater. Respiratory tract infection, 2.5 ml subcutaneously daily for 2 days, total 5 ml; aerosolized infection, 2.5 ml daily, total 5 ml; orally after brushing teeth, same dose as above (brushing teeth must cause bleeding from the dental bed). Semen inoculation, subcutaneous injection . 0.6 ml per day, 1.2 ml total; or intravaginal inoculation, 1.2 ml per day, 3.6 ml total; or delivery of semen deep into the vagina with a catheter and stimulation of the vagina with a finger cuff, resulting in damage to the vaginal mucous membranes and a bloody discharge.
C. Rhesus monkeys: males, 2 kg, 1-2 yr old, negative for HAV, HBV, HCV. Before inoculation with HBV, monkeys are irradiated with a 1-2 Cy dose of 60Co, WBC less than 4 x 109/L, and then a single intravenous injection of 2 ml of human HBV-positive serum is administered.
D. Rhesus monkeys: weighing 0.8-4 kg, 0.5-3 years old, and negative for HBV markers. 1 ml of fresh serum from patients with acute or chronic hepatitis B or serum from patients with chronic hepatitis B frozen for 2 years should be injected intravenously or intraperitoneally; HBV-positive serum (3-5 ml/kg) should be injected intravenously at one time.
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