Application of anti-thymocyte globulin in pretreatment of donor liver

Abstract

The invention relates to the field of biological medicine, and discloses an application of anti-thymocyte globulin in donor liver pretreatment, the application of the anti-thymocyte globulin in donor liver can effectively reduce the proportion of donor lymphocytes in a recipient after liver transplantation by inducing T cell failure, and the donor lymphocyte proportion can be effectively reduced. According to the present invention, with the application of the anti-thymocyte globulin, the occurrence of the graft-versus-host disease can be reduced, such that the effect of preventing the liver transplantation post-operation plant-versus-host disease can be achieved, the new application of the anti-thymocyte globulin in the preparation of the liver transplantation post-operation plant-versus-host disease can be determined based on the new property of the anti-thymocyte globulin, and the clinical indications of the anti-thymocyte globulin can be expanded; the internal principle that anti-thymocyte globulin participates in prevention of liver transplantation post-transplantation plant versus-host is disclosed, the curative effect is remarkable, and the application has outstanding popularization prospects.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine, and particularly relates to an application of anti-thymocyte globulin in pretreatment of donor liver. Background Art

[0002] Acute graft-versus-host disease (GVHD) after liver transplantation is a life-threatening complication of liver transplantation, with an incidence of approximately 1% and a mortality rate of up to 85%. The pathogenesis primarily results from sensitization and activation of donor T lymphocytes upon recognition of the host's MHC, leading to a cellular immune response against recipient tissues. This disease can develop systemically, affecting organs such as the skin, gastrointestinal tract, and bone marrow. Currently, there are no effective methods for preventing GVHD after liver transplantation, both domestically and internationally. Summary of the Invention

[0003] To address the deficiencies mentioned in the above-mentioned background technology, the present invention aims to provide an application of anti-thymocyte globulin in the pretreatment of donor livers, thereby effectively reducing the proportion of donor lymphocytes in the recipient's body after liver transplantation and reducing the occurrence of graft-versus-host disease, thereby preventing graft-versus-host disease after liver transplantation. Based on the new properties of anti-thymocyte globulin, its new use for preparing a drug for preventing graft-versus-host disease after liver transplantation has been determined.

[0004] The purpose of the present invention can be achieved through the following technical solutions: The invention discloses an application of anti-thymocyte globulin in pretreatment of donor liver. The anti-thymocyte globulin is dissolved in a carrier and then perfused into the donor liver through the portal vein. The dosage of the anti-thymocyte globulin is determined according to the weight of the donor liver.

[0005] Preferably, the dosage of the antithymocyte globulin is 20-25 mg / kg.

[0006] Preferably, the anti-thymocyte globulin is used by dissolving the anti-thymocyte globulin in sterile water for injection, and then dissolving the anti-thymocyte globulin in UW solution, and perfusing the solution into the liver through the portal vein.

[0007] Preferably, the anti-thymocyte globulin is used by dissolving the anti-thymocyte globulin in sterile water for injection, and then dissolving the anti-thymocyte globulin in HTK solution, and perfusing the solution into the liver through the portal vein.

[0008] Preferably, the perfusion rate is 100 mL / min.

[0009] Preferably, the donor liver is maintained completely immersed in the perfusion solution at 0-4°C during the perfusion.

[0010] Beneficial effects of the present invention: The present invention discloses an application of anti-thymocyte globulin in pretreatment of donor liver, including dosage and usage of the anti-thymocyte globulin. By pretreating the donor liver in vitro with the anti-thymocyte globulin, graft-versus-host disease can be effectively prevented. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, for ordinary technicians in this field, other drawings can be obtained based on these drawings without any creative work.

[0012] Figure 1 The donor liver of the present invention is pretreated with anti-thymocyte globulin, and CD3, CD4, and CD8 positive lymphocytes are detected by immunohistochemistry before and after the treatment. DETAILED DESCRIPTION

[0013] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts shall fall within the scope of protection of the present invention.

[0014] Example (before and after ATG) (1) After the donor liver is isolated, rice-sized liver tissue is obtained and cryopreserved (pre-ATG liver tissue, n=10). Anti-thymocyte globulin is dissolved in 10 mL of sterile water for injection, and then dissolved in 2000 mL of UW solution. The liver is perfused through the portal vein. The dosage of anti-thymocyte globulin is calculated based on the weight of the donor liver. The calculation method is 20-25 mg / kg. The perfusion rate is 100 mL / min. After perfusion, rice-sized liver tissue is obtained and cryopreserved (post-ATG liver tissue, n=10). (2) The frozen liver tissue was fixed, paraffin-embedded, and sliced. Paraffin-embedded liver tissue sections (4-5 μm) were taken and baked at 60°C for 30 min to fix the sections. The sections were then immersed in xylene I / II for 10 min each for dewaxing, hydrated with gradient ethanol (100% to 70%), and rinsed with distilled water. (3) Incubate in 3% H2O2-methanol solution at room temperature in the dark for 15 minutes to block endogenous peroxidase, then wash three times with PBS. The key step is antigen repair: immerse the slices in preheated 0.01M citrate buffer, microwave on medium heat until boiling and maintain for 15 minutes, cool naturally and rinse with PBS, add 5-10% normal goat serum to block at room temperature for 30 minutes, pour off the excess serum, and add primary antibodies to cover the tissues: mouse anti-human CD3 (1:100-1:200), CD4 (1:50-1:100), CD8 (1:50-1:100), incubate in a wet box at 4°C overnight or at 37°C for 2 hours, wash thoroughly with PBS three times, then add HRP-labeled anti-mouse secondary antibody, react at 37°C for 30 minutes, and wash thoroughly with PBS; (4) Use freshly prepared DAB solution for color development and monitor the dynamics under a microscope (about 1-3 minutes). When the lymphocyte membrane / cytoplasm shows brown-yellow particles, stop immediately with distilled water. Counterstain the cell nucleus with hematoxylin for 1 minute. After hydrochloric acid and alcohol differentiation, return to blue in running water. Dehydrate with gradient ethanol, make it transparent with xylene, and seal with neutral gum.

[0015] Experimental results The liver tissues obtained before and after ATG in the embodiment were experimentally verified: See Figure 1 ,Depend on Figure 1 It can be seen that the present invention uses anti-thymocyte globulin pretreatment of the donor liver, and immunohistochemistry is used to detect CD3, CD4, and CD8 positive lymphocytes in the donor liver tissue before and after treatment. It was found that the CD3, CD4, and CD8 positive lymphocytes decreased, indicating that anti-thymocyte globulin pretreatment can reduce the number of lymphocytes in the donor liver.

[0016] Throughout this specification, references to terms such as "one embodiment," "example," or "specific example" indicate that the specific features, structures, materials, or characteristics described in conjunction with that embodiment or example are included in at least one embodiment or example of the present invention. In this specification, schematic representations of these terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.

[0017] The basic principles, main features, and advantages of the present invention are shown and described above. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The above embodiments and descriptions are merely illustrative of the principles of the present invention. Various changes and modifications may be made to the present invention without departing from the spirit and scope of the present invention, and such changes and modifications fall within the scope of the invention as claimed.

Claims

1. An application of antithymocyte globulin in donor liver pretreatment, characterized in that: The anti-thymocyte globulin is dissolved in a carrier and then perfused into the donor liver through the portal vein. The dosage of the anti-thymocyte globulin is determined according to the weight of the donor liver.

2. The use of antithymocyte globulin in donor liver pretreatment according to claim 1, characterized in that: The dosage of the antithymocyte globulin is 20-25 mg / kg.

3. The use of antithymocyte globulin in donor liver pretreatment according to claim 1, characterized in that: The anti-thymocyte globulin is used by dissolving the anti-thymocyte globulin in sterile water for injection, then dissolving it in UW solution, and perfusing it into the liver through the portal vein.

4. The use of antithymocyte globulin in donor liver pretreatment according to claim 1, characterized in that: The anti-thymocyte globulin is used by dissolving it in sterile water for injection, then dissolving it in HTK solution, and then perfusing it into the liver through the portal vein.

5. Use of the antithymocyte globulin according to claim 3 or 4 in donor liver pretreatment, characterized in that: The perfusion rate was 100 mL / min.

6. Use of the antithymocyte globulin according to claim 3 or 4 in donor liver pretreatment, characterized in that: During the perfusion period, the donor liver is maintained completely immersed in the perfusion solution at 0-4°C.

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