Dendritic cell albumin suspension, preparation method and application

Abstract

The invention belongs to the technical field of immune effect drugs, and discloses a dendritic cell albumin suspension and a preparation method and application thereof.The preparation method of the dendritic cell albumin suspension includes the following steps that isolated mononuclear cells PBMC are collected, and matured DCs are cultured in a laminar flow purification laboratory; loading the multi-valence hepatitis B virus antigen epitope peptide on the DC; and the DC cell albumin suspension is prepared. The CTL immune effect excited by the DC can effectively remove HBV viruses in an integrated state in liver cells, so that the conditions of patients with hepatitis B and cirrhosis are reversed until HBV antigens are negative, and the conditions are relieved fundamentally; in the immune response excited by DC, the effector CTL cells immediately play a role in clearing HBV viruses, and the memory T lymphocytes play a role in clearing HBV viruses in vivo for a long time, so that the condition of hepatitis B is continuously reversed. Reagents used in the DC culture process are completely localized. The cost is greatly reduced and is obviously lower than that of foreign countries, and popularization is facilitated; china gets rid of the history of long-term dependence on imported anti-hepatitis B viruses.

Description

Technical Field

[0001] The invention belongs to the technical field of immune effect drugs, and in particular relates to an application of dendritic cell albumin suspension, a preparation method and an application thereof. Background Art

[0002] Among all types of hepatitis, hepatitis B is the most harmful.

[0003] Chronic hepatitis B is a chronic infection of the hepatitis B virus that leads to chronic inflammation of the liver tissue, which then develops into liver fibrosis, cirrhosis and even liver cancer. The high incidence of liver cancer in recent years is the result of the hepatitis B virus not being effectively eliminated from the body.

[0004] The conventional treatment plan for hepatitis B virus and liver cirrhosis according to the guidelines is: 1. Mainly use chemically synthesized nucleoside antiviral drugs; 2. Liver protection and enzyme reduction drugs; 3. CIK cell immunotherapy:. The defects of the existing technology are as follows: 1. Nucleoside antiviral drugs can only reduce the virus in the blood, and cannot eliminate the virus in an integrated state in the liver cells. The medication cannot be stopped during the medication process. Once stopped, the virus will rebound and it is very easy to develop drug resistance. 2. Liver protection and enzyme reduction drugs: only repair the liver cell membrane damaged by the virus. It cannot eliminate the virus in the liver cells and prevent the virus from damaging the liver cells. 3. Although the number of CIK cells is large, due to the lack of specific and targeted immune effects on the hepatitis B virus, its efficacy is limited.

[0005] At present, there is no specific drug in the world that can cure hepatitis B and its complications. Existing chemically synthesized nucleoside antiviral drugs and traditional Chinese medicine preparations can only inhibit the virus in the blood, but cannot completely eliminate the hepatitis B virus in an integrated state in liver cells, nor can they completely block the pathological process of hepatitis B patients developing into liver fibrosis, cirrhosis or even liver cancer.

[0006] The emergence of the dendritic cell (DC) theory has shown a bright future for immunotherapy of hepatitis B cirrhosis. Dendritic cells are the most powerful antigen-presenting cells in the human body and the primary cells of immune response. They participate in antigen recognition, processing and presentation, can effectively stimulate T cell responses and induce the production of specific cytotoxic T lymphocytes, and initiate, maintain and regulate immune responses. DC-based antiviral and anti-tumor immunotherapy has become a research hotspot.

[0007] Dendritic cell immunotherapy can treat diseases related to viral infection such as hepatitis B and all solid tumors. The dendritic cell and receptor theory won the Nobel Prize in Medicine on October 3, 2011. This is a great discovery and a major contribution to medical immunology. It has opened up a new and reliable treatment path for humans to overcome viral infection-related diseases such as hepatitis B and cancer!

[0008] The main reason why chronic hepatitis B and liver cirrhosis cannot be cured is that the human immune system develops "immune tolerance" to the hepatitis B virus (HBV). Modern research has found that the main reason for immune tolerance to hepatitis B is the functional deficiency of "dendritic cells (DC)". DC does not recognize the infected hepatitis B virus, and cannot transmit the hepatitis B antigen information to T lymphocytes, so it cannot stimulate the immune clearance effect against the hepatitis B virus, and coexists peacefully with the virus, and then unknowingly and slowly damages liver cells and develops cirrhosis or even liver cancer. In order to break immune tolerance, effectively reverse the hepatitis B condition, and prevent the development of hepatitis B, we have successfully treated more than 7,000 hepatitis B patients with "dendritic cell (DC) immunotherapy" and reversed the patients' condition. Specifically, the mononuclear cells (PBMC) of the patient or close relatives are collected, a normal functioning DC cell is cultured, and loaded with "multivalent hepatitis B virus dominant antigen epitope peptides", just like a missile is built to input enemy target information. The mature DC cells loaded with HBV antigen peptide information are called "cell missiles" and are infused back into the patient's body in batches. This can stimulate a specific and targeted immune clearance effect against the hepatitis B virus (HBV), which can not only clear the HBV virus in the blood, but more importantly, it can clear the virus in an integrated state in the nucleus of liver cells, fundamentally reversing the condition of patients with hepatitis B and cirrhosis and achieving a cure.

[0009] Through the above analysis, the problems and defects of the existing technology are: there is no effective treatment for the basic cause of immune tolerance of hepatitis B - "dendritic cell (DC) dysfunction", which leads to the long-term incurable hepatitis B and the progression of the disease. The symptoms are treated but not the root cause, which makes the hepatitis B disease continue to develop towards cirrhosis and liver cancer. Only by cultivating a kind of "dendritic cell (DC)" with normal function, and loading "hepatitis B virus (HBV) dominant antigen epitope peptide" on its surface MHC receptor, and loading DC cells with accurate HBV target information, and then returning it to the patient, the TCR receptor of T lymphocytes accepts the HBV antigen information transmitted by DC, which can stimulate the CTL immune clearance effect of hepatitis B virus inside and outside the blood and liver cells in the body, and can reverse the hepatitis B and cirrhosis and prevent the development towards liver cancer. Summary of the invention

[0010] In view of the problems existing in the prior art, the present invention provides a dendritic cell (DC) albumin suspension loaded with "HBV antigen epitope peptide", a preparation method and application thereof.

[0011] The present invention is achieved by using a dendritic cell (DC) albumin suspension loaded with "HBV antigen epitope peptides", comprising:

[0012] Furthermore, the application of the albumin suspension of dendritic cells (DC) loaded with "HBV antigen epitope peptide" stimulates a specific and targeted CTL immune effect against hepatitis B virus, and can effectively eliminate HBV viruses in an integrated state in the blood and liver cell nuclei.

[0013] Furthermore, the specific immune response immune effect stimulated by the application of the dendritic cell (DC) albumin suspension loaded with "HBV antigen epitope peptide" will result in an increase in transaminase and HBV-DNA, a unique immune response reaction, when clearing the integrated virus in the liver cells.

[0014] Furthermore, the application of the dendritic cell (DC) albumin suspension can not only remove the HBV virus in the blood, but also, more importantly, can remove the virus in an integrated state in the nucleus of liver cells, thus reversing the condition of hepatitis B liver cirrhosis.

[0015] Another object of the present invention is to provide a method for preparing the dendritic cell (DC) albumin suspension, the method for preparing the dendritic cell albumin suspension comprising the following steps:

[0016] The first step is to use a German-made blood cell separator to collect 20-30 ml of PBMC from the patient or his / her biological children.

[0017] The second step is to culture mature DCs in a laminar flow cleanroom and load DCs with multivalent HBV antigen epitope peptides;

[0018] The third step is to prepare DC cell albumin suspension.

[0019] Another object of the present invention is to provide a preparation for immunological treatment of hepatitis B virus, namely, a dendritic cell (DC) albumin suspension loaded with "HBV antigen epitope peptide"

[0020] Another object of the present invention is to provide an immunological therapeutic drug for treating liver cirrhosis, wherein the immunological drug for treating liver cirrhosis comprises the dendritic cell albumin suspension.

[0021] In combination with the above technical solutions and the technical problems solved, the advantages and positive effects of the technical solutions to be protected by the present invention are as follows:

[0022] First, dendritic cells (DC) are important cells in the primary link of the human immune response against viruses and tumor cells. Studies have shown that the main reason why hepatitis B cannot be cured for a long time and immune tolerance occurs is the loss of DC function. Cultivating a functional DC cell can stimulate the body's specific and targeted immune effect against hepatitis B virus, so as to clear the blood and, more importantly, the virus in an integrated state in the nucleus of liver cells, so as to reverse the condition of patients with hepatitis B and liver cirrhosis and prevent the development of hepatitis B liver cancer.

[0023] The CTL immune effect stimulated by the DC of the present invention can effectively remove the HBV virus in the integrated state in the liver cells, so that the condition of patients with hepatitis B and liver cirrhosis is reversed until the HBV antigen turns negative, and the condition is fundamentally alleviated; the immune response stimulated by DC, in which the "effect CTL cells" immediately play a role in removing the HBV virus, and the "memory T lymphocytes" will play a long-term immune effect of removing the virus. When the hepatitis B treatment is stopped to a certain extent, this memory immune effect will continue to play a therapeutic role, so that the condition will not rebound and relapse, thereby achieving the purpose of preventing and stopping the occurrence of liver cirrhosis and liver cancer; DC treatment has completely changed the current conventional treatment situation that CIK cells and nucleoside drugs cannot remove the virus in the integrated state in the liver cells, and stopping the drug and stopping the treatment will inevitably cause the condition to rebound. The reagents used in the process of culturing DC are completely domestically produced. The cost is greatly reduced, and the cost is significantly lower than that in foreign countries, which is convenient for promotion; it has enabled my country to get rid of the history of long-term dependence on imported anti-hepatitis B virus.

[0024] Second, the present invention uses three intravenous infusions and subcutaneous injections around the axillary lymph nodes in order to stimulate specific and targeted CTL immune effects against hepatitis B virus, thereby achieving the purpose of clearing HBV virus in the blood and liver cell nuclei in an integrated state. The "multivalent hepatitis B virus antigen epitope peptide" is synthesized to provide DC cells with accurate target information, so that the stimulated CTL immune effect of clearing HBV in the liver is specific and targeted; an "immunopotentiator PL" is used in the process of culturing DC cells, which significantly enhances the cell phenotype of DC and significantly increases the release of endogenous interferon; the invention is the first to collect children's mononuclear cells (PBMC) to culture DC cells, and then infuse them back to sick parents. The immune efficacy is stronger than that of DC cultured with the patient's own cells, which accelerates the improvement of the disease.

[0025] Third, as auxiliary evidence of the inventiveness of the claims of the present invention, it is also reflected in the following important aspects:

[0026] (1) The expected benefits and commercial value of the technical solution of the present invention after transformation are as follows: DC cell immunotherapy has overcome the global medical problem that my country has long lacked an effective treatment for hepatitis B cirrhosis. It not only provides hospitals at all levels with an effective method for treating hepatitis B cirrhosis, benefiting a large number of hepatitis B cirrhosis patients, but also increases the clinical benefits of hospitals and drives the production of downstream cell culture equipment, various reagents and cytokine production companies. Its commercial value is difficult to estimate.

[0027] (2) The technical solution of the present invention fills the technical gap in the industry at home and abroad: it changes the harsh situation of hepatitis B patients who have long relied on nucleoside antiviral drugs, rebound after stopping the drugs, and cannot reverse the hepatitis B disease.

[0028] (3) The technical solution of the present invention solves the technical problem of hepatitis B treatment that people have been eager to solve but have never been successful. It also solves the problem that people have been eager to have an effective and feasible solution to effectively treat hepatitis B and liver cirrhosis and reverse the disease, and achieves a breakthrough in the revolutionary treatment of hepatitis B and liver cirrhosis.

[0029] (4) The technical solution of the present invention overcomes technical prejudice: The technical solution of the present invention breaks the current situation of relying solely on drugs to treat hepatitis B, breaks the textbook dogma that cirrhosis is irreversible, and successfully reverses hepatitis B cirrhosis to the morphology of a normal liver. DC cell immunotherapy has reversed the condition of a patient with severe cirrhosis who required liver transplantation, verifying the wise conclusion of Academician Wu Zuze that "cell therapy will be the ultimate treatment for viral infection-related diseases and tumors." BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 It is a flow chart of a method for preparing a dendritic cell albumin suspension provided by an embodiment of the present invention;

[0031] Figure 2 It is a schematic diagram of the CTL immune effect stimulated by DC provided in an embodiment of the present invention;

[0032] Figure 3 Schematic diagram of the frequency distribution of IFN-γ-secreting CTL cells activated by P-DC dendritic cells provided in an embodiment of the present invention. DETAILED DESCRIPTION

[0033] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0034] like Figure 1 As shown, the method for preparing a dendritic cell albumin suspension provided in an embodiment of the present invention comprises the following steps:

[0035] S101: Use German blood cell separator to collect 20-30ml of mononuclear cells (PBMC) from the patient or his / her biological children;

[0036] S102: Cultivate mature DCs in a laminar flow cleanroom and load them with “multivalent hepatitis B virus antigen epitope peptides”;

[0037] S103: preparing DC cell albumin suspension.

[0038] The dendritic cell albumin suspension provided in the embodiment of the present invention is intravenously reinfused three times and subcutaneously injected around the axillary lymph nodes to stimulate specific and targeted CTL immune effects against hepatitis B virus, thereby achieving the purpose of clearing HBV virus in an integrated state in the blood and liver cell nuclei.

[0039] The present invention has successfully synthesized the key substance of hepatitis B virus infection: "multivalent hepatitis B virus dominant antigen epitope peptide", which provides accurate targeting information for DC cell immunotherapy to treat chronic hepatitis B. The present invention calls the DC cells loaded with "hepatitis B virus dominant antigen epitope peptide" T-DC. T-DC has been used to successfully treat more than 7,000 hepatitis B patients, including patients with liver cirrhosis, and significantly reversed the patients' condition. The clinical application of dendritic cell (DC) immunotherapy was appraised by an expert group headed by Academician Wu Zuze in September 2005. The experts unanimously gave high praise and affirmation to the clinical application of DC immunotherapy.

[0040] Typical cases of treating viral hepatitis, liver cirrhosis, and tumors are listed below. From them, we can see that DC immunotherapy effectively curbs and eliminates hepatitis viruses, which is different from conventional drug treatments! It is manifested in the following rules and characteristics:

[0041] 1. In the process of DC treatment for chronic hepatitis B and liver cirrhosis, transaminase and HBV-DNA may increase, and they can return to normal without liver protection, enzyme reduction and antiviral drugs. This is the manifestation of the CTL immune response stimulated by DC, which effectively clears the integrated virus in the liver cells. This clearance effect cannot be achieved by any antiviral drug. The result of clearing the integrated virus in the liver cells is the reversal of the disease, the normalization of liver function, and especially the significant increase of albumin in patients with liver cirrhosis.

[0042] 2. The rule of globulin changes: During the treatment, while albumin rises, globulin first decreases, but then increases in the later stage. This is a unique rule of DC cell therapy; the virus is cleared in the early stage, liver function improves, and aging globulin is processed, so globulin decreases. In the later stage, because DC induces B lymphocytes to produce new antibodies (globulins) through CD4+T lymphocytes, the globulin of patients receiving DC treatment will increase in the later stage. Although the albumin-globulin ratio decreases, the overall condition is improving. Therefore, patients receiving DC immunotherapy cannot mechanically view the changes in the condition based on the high or low albumin-globulin ratio.

[0043] 3. About the memory effect of DC-CTL: DC treatment is not like antiviral drugs that can only inhibit viruses in the blood, and the condition will generally rebound after stopping the drug. The CTL immune effect stimulated by DC not only has targeted, specific, and active immune killing and clearing of viruses in liver cells, bringing improvement in various liver function indicators, but also this immune effect has a long-term memory function. When the treatment is stopped for many years, the results of regular observations have proved that this CTL effect continues to play a role, and various clinical indicators continue to improve after stopping treatment.

[0044] 4. After treatment with DC cell immunotherapy, patients often show significant improvement in complexion, mental and physical strength, enhanced immune function (such as being less susceptible to colds, and unexpected disappearance of combined HPV, HSV and other pathogen infections), and complexion changes from dull to rosy; the color of liver palms and spider nevi gradually lightens until they disappear.

[0045] Table 1 Expansion and phenotypic identification of P-DC and the effect of PL enhancer on their surface molecular expression

[0046]

[0047] Case 1: Su**, male, 51 years old, had hepatitis B and liver cirrhosis for more than 10 years and received 6 courses of treatment. He had been treated with adefovir for 7 years. In the middle and late stages of DC treatment, HBV-DNA increased while transaminase increased. This was a manifestation of the immune response CTL effectively clearing the virus in the cells, and the overall liver cirrhosis condition was significantly reversed.

[0048] Table 2

[0049] Check time 2013-3-23 2013-9-23 2013-10-23 2013-12-28 2014-6-26 2014-9-9 HBV·DNA ＜1000 <![CDATA[6.34×10 6 ]]> <![CDATA[1.86×10 7 ]]> <![CDATA[1.89×10 5 ]]> ＜1000 ＜1000 <![CDATA[ HBsAg ]]> 6276 882 648 471 289 241 ALT 58 45 197 134 26 23 AST 39 76 191 462 41 35

[0050]

[0051] Case 2: Song**, male, 44 years old, had early liver cirrhosis for nearly ten years. In the middle and late stages of DC treatment, while transaminase increased, HBV-DNA also increased, which was a manifestation of the immune response CTL effectively clearing the virus in the cells. The overall condition was also obviously improving, and HBsAg finally turned negative during the observation period.

[0052] Table 3

[0053]

[0054] Case 3 Hu*, who had suffered from hepatitis C cirrhosis and hepatogenic diabetes for more than ten years, received five courses of treatment. His liver function improved significantly, HCV-RNA turned negative, and his blood sugar level dropped significantly, eventually returning to normal. Ultrasound showed improvement in liver spots and narrowing of the portal vein.

[0055] Table 4

[0056]

[0057] Case 4: Patient Lu** was diagnosed with Hepatitis B virus (HBV) for one year and had been treated with lamivudine for one year, but the results were not good. He started DC treatment on December 25, 2009. After each treatment, he was not only re-examined in this hospital, but also had a second blood test at the Affiliated Hospital of Medical University on the same day. After stopping treatment, he turned negative during the observation period, and then he showed the test results from Medical University to the doctor, telling him that the test results of the two hospitals were basically the same.

[0058] Table 5

[0059] Check time HBV-DNA (cps / ml) HBsAg (ng / ml) 2009-12-25 <![CDATA[8.168×10 6 ]]> 6917 2010-2-3 <![CDATA[4.92×10 6 ]]> 3899 2010-3-5 (stop taking medication) <![CDATA[1.29×10 7 ]]> 1902 2010-4-4 <![CDATA[4.11×10 7 ]]> 1283 2010-5-3 <![CDATA[7.6×10 5 ]]> 874 2010-6-2 <![CDATA[3.07×10 5 ]]> 658 2010-8-2 <![CDATA[8.6×10 4 ]]> 481 2010-11-24 drug discontinuation) <![CDATA[3.09×10 4 ]]> 181

[0060] Table 6

[0061] Observation period 2011-3-18 2011-5-15 2011-7-20 HBV-DNA <![CDATA[4.03×10 3 ]]> ﹤1000 ﹤1000 HBsAg 181 108 ﹤10

[0062] Table 7

[0063]

[0064] The results of the examinations at the First Affiliated Hospital of Medical University during the same period were basically the same.

[0065] Table 8

[0066] Check time HBV-DNA HBsAg HBsAb 2010-9-26 <![CDATA[7.35×10 4 ]]> ﹥225 2011-3-16 <![CDATA[3.48×10 4 ]]> 16.84 2011-6-24 ﹤1000 1.02 2011-7-21 ﹤1000 0.5 3.4

[0067] Table 9

[0068]

[0069] Case 5, Liu**, female, 66 years old, hepatitis B virus (HBV) positive, HBsAg turned negative during the observation period after stopping treatment. Her son asked her mother to undergo reexamination and verification in multiple hospitals, and the results of the verification in multiple hospitals were consistent.

[0070] Table 10

[0071]

[0072] Table 11

[0073] B-ultrasound 2008-9-11 2008-12-31 2009-8-30 Liver parenchyma Light spot thickening Slightly thickened Fine distributed Lack of uniformity Uniform Uniform Spleen thickness 2.8㎝ 2.2㎝ 2.2㎝

[0074] After three years of follow-up observation by Guilin People's Hospital, the First Affiliated Hospital of Traditional Chinese Medicine, and Guilin 181 Hospital, the patient's liver function was normal, and blood sugar, blood lipids, and blood rheology were normalized. He had stopped taking blood sugar, blood lipid, and blood pressure lowering drugs. Since DC cell immunotherapy effectively cleared the hepatitis B virus in liver cells, liver function improved, sugar and fat metabolism was normal, blood rheology and blood pressure also tended to be normal, and he stopped taking drugs for the treatment of three highs for many years and recovered.

[0075] Case 6 Li**, male, 58 years old, advanced liver cirrhosis, with significantly decreased serum albumin, lower limb edema, trace ascites, and then developed to bedridden. He was admitted to Xi'an Infectious Disease Hospital and received antiviral liver protection treatment for half a year without improvement. Then he had upper gastrointestinal bleeding and underwent portal vein-caval shunt surgery in Xijing Hospital, which caused hepatic encephalopathy. He was then transferred to the First Affiliated Hospital of Dalian University for 14 days of hospitalization and prepared for liver transplantation. While waiting for the opportunity of liver source, he was introduced to receive "dendritic cell (DC) immunotherapy" treatment. After three courses of DC cell culture with mononuclear cells (PBMC) from his children, his condition was significantly reversed, his mental and physical strength were enhanced, and he did not need liver transplantation. After another course of intensive treatment, various laboratory indicators improved significantly, and the surface antigen had turned negative. The various laboratory indicators are shown in Table 12.

[0076] Table 12

[0077] Time Project 2020.10.20 2021.6.23 2022.2.26 WBC 2.5 3.5 4.33 L- 0.5 0.8 0.83 RBC 3.56 3.89 3.87 Hb 127 136 139 PLT 37 38 52 ALT 38.3 36.65 32.4 AST 44.95 53.08 45.2 ALB 34.41 29.85 37 GIO 26.48 26.19 23 A / G 1.3 1.14 1.61 Sag 0.77 0.18 0 HBV-DNA <![CDATA[<10 2 ]]> <![CDATA[<10 2 ]]> <![CDATA[<10 2 ]]>

[0078] Case 7: Patient Li*, who was born with chronic hepatitis B and had cirrhosis for more than ten years, was often weak, had poor appetite, and was in a very bad mental state. Many abnormalities were found in liver function and liver stiffness tests. After three courses of DC cell immunotherapy, he is now in the observation period. Since DC therapy can effectively remove the integrated virus in the nucleus of liver cells, the condition of liver cirrhosis is reversed, as shown by a significant increase in albumin from 34.8 to 51.5; a decrease in total bile acid (TBA): 171.9-9.4; CHE 7189-12175HBV-DNA from 3.03x105 to 5.59x103. The increase in HBsAg is the result of the virus being cleared and released from the cells into the blood. The non-invasive test of liver stiffness dropped from 28.1 to 6.6, which is the result of DC immunotherapy removing the virus in the liver cells, reversing the condition of liver cirrhosis. The results of the examination are summarized in Table 13.

[0079] Table 13

[0080]

[0081] From the above cases, we can see that the specific immune response effect stimulated by DC, when clearing the integrated virus in liver cells, is manifested as an increase in transaminase and HBV-DNA. This is a unique therapeutic effect of DC immunotherapy, which is the result of the elimination of viral genes from the nucleus to the outside. Without antiviral and liver-protecting and enzyme-lowering drugs, all indicators can return to normal, and the patient's condition can be reversed!

[0082] DC cell immunotherapy, known as "cell missiles", can stimulate an active, targeted and specific immune clearance effect against hepatitis B virus (HBV). It can not only clear the HBV virus in the blood, but more importantly, it can clear the virus in an integrated state in the nucleus of liver cells. This treatment also has a memory effect and continues to exert a therapeutic effect during the observation period after cessation of treatment. Ultimately, it can fundamentally reverse the condition of patients with hepatitis B and cirrhosis and achieve a cure.

[0083] The above description is only a specific implementation mode of the present invention, but the protection scope of the present invention is not limited thereto. Any modifications, equivalent substitutions and improvements made by any technician familiar with the technical field within the technical scope disclosed by the present invention and within the spirit and principle of the present invention should be covered by the protection scope of the present invention.

Claims

1. A key substance for hepatitis B virus infection, characterized in that: Multivalent hepatitis B virus dominant antigen epitope peptides.

2. A DC cell loaded with the hepatitis B virus dominant antigen epitope peptide of claim 1 for treating liver cirrhosis.

3. A method for using a dendritic cell albumin suspension, characterized in that: The application of the dendritic cell albumin suspension stimulates a specific and targeted CTL immune effect against the hepatitis B virus, thereby clearing the HBV virus in an integrated state in the blood and liver cell nuclei.

4. The use of dendritic cell albumin suspension according to claim 1, characterized in that: The specific immune response immune effect stimulated by the application of dendritic cell albumin suspension to stimulate DC will manifest as the unique phenomenon of increased transaminase and increased HBV-DNA when clearing the integrated virus in the liver cells.

5. The use of dendritic cell albumin suspension according to claim 1, characterized in that: The dendritic cell albumin suspension is used to remove HBV virus in the blood and remove the virus in an integrated state in the nucleus of liver cells.

6. A method for preparing a dendritic cell albumin suspension according to any one of claims 1 to 4, characterized in that: The preparation method of the dendritic cell albumin suspension is described, The following steps are involved: The first step is to collect ex vivo mononuclear cells (PBMCs) and culture mature DCs in a laminar flow purification laboratory. In the second step, DCs are loaded with multivalent HBV epitope peptides; The third step is to prepare DC cell albumin suspension.

7. An immune drug for treating hepatitis B virus, characterized in that: The immune drug for treating hepatitis B virus comprises the dendritic cell albumin suspension as described in any one of claims 1 to 4.

8. An immune drug for treating liver cirrhosis, characterized in that: The immune drug for treating liver cirrhosis comprises the dendritic cell albumin suspension as described in any one of claims 1 to 4.

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