WO2006047954A1 - Procédé d'obtention et d'application des matières pour la reconnaissance immunologique d'autoantigènes - Google Patents

Procédé d'obtention et d'application des matières pour la reconnaissance immunologique d'autoantigènes Download PDF

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WO2006047954A1
WO2006047954A1 PCT/CN2005/001842 CN2005001842W WO2006047954A1 WO 2006047954 A1 WO2006047954 A1 WO 2006047954A1 CN 2005001842 W CN2005001842 W CN 2005001842W WO 2006047954 A1 WO2006047954 A1 WO 2006047954A1
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cells
immune
immune recognition
recognition
stem cells
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PCT/CN2005/001842
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Chinese (zh)
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Ziyi Cheng
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Ziyi Cheng
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/001—Preparations to induce tolerance to non-self, e.g. prior to transplantation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators

Definitions

  • the present invention relates to a method and method for obtaining raw materials required for re-immunization of self-antigens after birth by treating rejection and immune system diseases.
  • an early immune recognition hematopoietic stem cell and/or a late immune recognition hematopoietic stem cell required for re-imaging the self-antigen immune recognition and a tissue organ related to immune recognition at the time of obtaining an autoantigen immune recognition period, including: thymus Cells and factors related to immune recognition of autoantigens in the bone marrow.
  • a treatment method for re-imaging the autoantigen by recognizing the tissues and organs and the graft antigens of the autoantigens in the body for the treatment of rejection and immune system diseases can not only treat a variety of diseases associated with diseases of the immune system and rejection reactions after organ transplantation, but also treat a variety of different organs and different types of diseases of the recipient.
  • the invention can be used not only in humans but also in animals. BACKGROUND OF THE INVENTION Noun annotations used in the present invention, and methods for obtaining and applying materials required -
  • the immune recognition period of self antigen is divided into -
  • early autoantigen immune recognition of hematopoietic stem cells is an immunologically recognized hematopoietic stem cell in the pre-immune recognition of autoantigens and in the autoimmune recognition phase of autoantigens.
  • the late immune recognition hematopoietic stem cells are immunologically recognized hematopoietic stem cells at the end of the autoantigen immune recognition.
  • Early immune recognition Hematopoietic stem cells may include: hematopoietic stem cells associated with immune recognition, and/or immune recognition, and/or group cells, etc., and/or in autoantigen immune recognition pre- and/or autoantigen immunological phase, and/or The differentiated cells, and/or the various differentiated mature blood system cells associated with immunity, early immune recognition hematopoietic cells are cells that can be self-immunologically recognized under certain circumstances.
  • hematopoietic pluripotent stem cells including Haematopoietic Stem Cells
  • various immune system-related progenitor cells immunoglobules, monocyte stem cells, precursor B cells, precursor T cells, megakaryocyte stem cells, and cells secreting autoantigen immune recognition factors, and/or secreting autoantigen immune recognition factors Cells, etc.
  • various immune-related cells are referred to in this specification as abbreviations: Early immune recognition of hematopoietic cells.
  • Late immune recognition Hematopoietic stem cells may include: stem cells that are associated with immune recognition and/or immune recognition, or differentiated immature cells, or various types of stem cells related to immunity, at the end of autoantigen immune recognition, And/or undifferentiated mature cells, late II immunoreactive hematopoietic stem cells are generally unable to carry out autoantigen immune recognition, only in special cases involving "organizations, organs, cells, factors related to autoantigen immune recognition" It is possible to carry out self-antigen immune recognition.
  • hematopoietic pluripotent stem cells including Haematopoietic Stem Cells
  • various immune-related progenitor cells immunological pluripotent stem cells
  • myeloid stem cells lymphoid stem cells
  • various Immune-related unipotent stem cells such as: granules, monocyte stem cells, precursor T cells (ie, proT and early proT cells), 'precursor B cells (ie, proB and early proB cells), monocytes (DC) megakaryocytes And so on and various immune-related cells
  • granules monocyte stem cells
  • precursor T cells ie, proT and early proT cells
  • 'precursor B cells ie, proB and early proB cells
  • monocytes DC megakaryocytes
  • immune-related cells such as: granules, monocyte stem cells, precursor T cells (ie, proT and early proT cells), 'precursor B cells (ie, proB and early proB cells), monocytes
  • Immune recognition of hematopoietic stem cells includes: early immune recognition of hematopoietic stem cells, and/or late immune recognition of hematopoietic stem cells.
  • immunologically recognized hematopoietic stem cells are also referred to as non-immunologically recognized stem cells, or immunologically recognized hematopoietic stem cells.
  • the tissues, organs, cells, and factors related to the autoimmune recognition of autoantigens are the "preimmune recognition of autoantigens", and/or the embryonic and fetal stages of human or animal immunization recognition period, early immune recognition Hematopoietic stem cells can perform self-antigen immune recognition period, participate in early immune recognition of hematopoietic stem cells, and/or tissues, and/or cells, and/or hormones related to immune recognition, which are immunologically recognized by autoantigens in the body. Cytokines, peptides, proteins, and more.
  • organs such as thymus, spleen, lymph nodes and bone marrow, and various cells and hormones contained in these organs.
  • various cytokines, polypeptide proteins, tissues, tissue fluids, cells, and the like See “Cellular and Molecular Immunology” Chapter 6, Chapters VII and IX, “Cellular and Molecular Immunology", Second Edition, Jin Boquan, Editor-in-Chief, China Science Press, 2001, first print).
  • Microorganisms, organs, cells, factors related to immune recognition of autoantigens may include:
  • the tissues, organs, cells, and genes involved in the immune recognition of autoantigens include: human or animal embryos and fetal stages, and participate in early immune recognition of hematopoietic stem cells to the body, organs, organs, organs, organs, A cell, a factor, a complete tissue, an organ, or a partially incomplete tissue or organ. It provides the microenvironment required for early immune recognition of hematopoietic stem cells to immunologically recognize autoantigens in the body, providing early immune recognition to recognize hematopoietic stem cells.
  • "Self-antigen immune recognition factor” may include : a starter factor, a recognition factor, a termination recognition factor, and various microenvironments such as a tissue fluid that provides a self-antigen for various auxiliary factors required for immunological recognition.
  • tissue, organ, cell, factor related to immune recognition of autoantigen may include all cells of the above-mentioned tissue and organs, and / or part of cells, and / or one or more of these special cells, into the body, part Cells can reach the target organ through homing, grow in the target organ, and grow and develop outside the target organ, providing the microenvironment needed for early immune recognition of hematopoietic stem cells to identify the autoantigen in the body.
  • Early immune recognition identifies cells that secrete "autoantigen immune recognition factor” required for hematopoietic stem cell immune recognition of autoantigens in the body, and/or cells that secrete "promoting (producing) autoantigen immune recognition factors”.
  • cytokines such as cells that secrete autoantigens for various cofactors required for immunological recognition, and various helper cells involved in immune recognition of self-antigens, and various micro-requirements for providing self-antigen for immunological recognition surroundings.
  • tissue, organ, cell, factor related to autoantigen immune recognition may include "self-antigen immunization required by the above-mentioned tissue and organ to provide early immune recognition for hematopoietic stem cells to recognize the autoantigen in the body.
  • the recognition factor ", and/or "promotes (secrets) autoantigen immune recognition factor”, and/or various cofactors required for immunological recognition of self-antigens, and the like. You can use the above several organizers as needed. One or more of several types of materials such as officials, cells, and factors.
  • the various factors used can be extracted by various methods of factor extraction currently used, and can be produced by various methods such as transgenic cell cloning.
  • Microorganisms, organs, cells, and factors related to autoantigen immune recognition are also referred to in this specification as: “immune recognition organs” or “initiation recognition factors” or “activation factors” or “suspension recognition factors” or “stop factors” ".
  • tissue and organs related to autoimmune antigen recognition there are various methods for finding tissues and organs related to autoimmune antigen recognition, including: using the method of Example 9 of the present invention, making tissues and/or organs undergoing self-antigen immune recognition into single cells. Suspension, or use a part of tissue organs, or use intact tissue organs, transplanted into animals separately or in different combinations, to carry out autoantigen immune recognition experiments, if any tissue, organ or combination of those tissues and organs can The self-antigen immune recognition experiment is successful, and the tissues and organs included in the tissue, the organ or the combination are "organisms and organs related to the immune recognition of the self antigen".
  • Example 9 of the present invention injection into a test animal alone or in a different combination, and performing an autoantigen immunoreactivity assay, if any protein, amino acid peptide chain (polypeptide) or those proteins, amino acid peptide chains (polypeptides)
  • the combination of these can make the self-antigen immune recognition experiment successful, and the protein, amino acid peptide chain (polypeptide) or the amino acid peptide chain (polypeptide) included in this combination is "a factor related to the immune recognition of the self antigen".
  • the culture fluid input into the body contains "immunity recognition related to autoantigen”. Factor”. Then, using chromatography and mass spectrometry, different proteins and amino acid peptide chains (polypeptides) between the two cells were found. Using the method of Example 9 of the present invention, it is injected into an experimental animal to perform an autoantigen immune recognition experiment. If the autoantigen immunological recognition experiment is successful, the protein and amino acid peptide chain (peptide) input into the body is "with Self-antigen immune recognition related factors".
  • graft Preparing transplants of organs, tissues, cells, etc. that have been transplanted into the body, and/or cells, tissues, organs, proteins, polypeptides, etc. of the antigenic family carrying the immune recognition, etc., in the present specification
  • An antigenic organ, tissue, cell, or the like such as a tissue, is used as a donor antigen donor for early immune recognition of hematopoietic stem cells and/or late immune recognition of hematopoietic stem cells for self-antigen recognition. These are referred to in the present specification as: graft antigens.
  • the autoantigen immune recognition factor "initiates, promotes, and terminates the self-immunological immune recognition factor", and/or “promotes, promotes, terminates Self-antigen immune recognition factors” and cells that secrete these factors, but they can be found through medical practice, and the presence of these factors and the tissues and organs of the cells secreting these factors are "related to the autoimmune recognition of autoantigens.”
  • Tissues, organs, cells, factors including: embryos from 3 months to 5 months or so in the thymus tissue of the autoantigen immune recognition phase, and/or bone marrow tissue, etc., may exist in these tissues and organs A cell that secretes a "autoantigen immune recognition factor", and/or a cell that secretes a "promoting (producing) autoantigen immune recognition factor”.
  • Each of these factors may be a single factor or a composite factor composed of a plurality of factors.
  • the activation of the autoantigen immune recognition factor, the promotion of the autoantigen immune recognition factor, and the termination of the autoantigen immune recognition factor may be a plurality of different factors, or the same factor may play different roles at different stages, and these factors may be: Hormones, cytokines, peptides, proteins, tissues, tissue fluids, cells, organs, and more. It has been discovered through medical practice that these factors can sometimes transform late-stage immune recognition hematopoietic stem cells to re-identify their own antigens.
  • an immune recognition microenvironment that provides input and promotes autoantigen immune recognition by inputting organs and tissues (such as thymus tissue, thymus cells, bone marrow tissue, bone marrow cells, etc.) that provide an immune recognition microenvironment that promotes autoantigen immune recognition. These factors have been introduced to promote the recognition of autoantigens.
  • a factor related to initiation, promotion of recognition of autoantigen immune recognition by an immune-free recognition cell, and termination of recognition and an organ and tissue (eg, thymus tissue) providing an immune recognition microenvironment for promoting autoantigen immune recognition , thymocyte and bone marrow tissue, bone marrow cells, etc.)
  • an organ and tissue eg, thymus tissue
  • initiation factor motivating recognition factor
  • termination recognition factor initiation factor
  • these factors are collectively referred to in this specification as: Startup recognition factor.
  • the relevant factors for terminating the non-immunologically recognized stem cells for self-antigen immune recognition may also be referred to in the present specification as a termination identifier.
  • Immune system diseases which may include, for example, systemic lupus erythematosus, rheumatism, rheumatoid arthritis, glomerulonephritis, scleroderma, etc., and may also include antigenic antigenic changes due to various causes of the disease.
  • the induced immune disease can cause changes in autoantigens, and the mutated autoantigens can be organs, tissues, cells, proteins, polypeptides, etc., which are referred to in the present specification as: mutated autoantigens.
  • Bone marrow transplantation The more popular word is "hematopoietic stem cell transplantation”.
  • the bone marrow transplantation treatment used in the present invention is an immunologically recognized hematopoietic stem cell transplantation by performing clearance of immune-related cells in the present invention, and/or performing "organisms, organs, cells, factors related to autoantigen immune recognition". "Transplantation, the treatment process for secondary self-antigen immune recognition.
  • the method of bone marrow transplantation is -
  • drugs such as: cyclophosphamide
  • radiation such as: cobalt 6 °, medical linear accelerator, high-energy X-ray systemic irradiation, etc.
  • immunological methods can also be used to remove specific cells, and/or inactivate cytokines, such as T lymphocytes and other immune cells, such as the use of CD3 monoclonal antibodies combined with cytotoxins to form cytotoxic drugs plus complement input.
  • the body kills T lymphocytes in the body and anti-lymphocyte globulin (ALG) and other drugs that kill immune cells in the body's immune system. This is a specific therapy for a certain cell or a certain type of cells. .
  • myeloablative treatment can also use partial myeloablative or non-myeloablative treatment. Then, hematopoietic stem cell transplantation is performed on the recipient, and the recipient is reconstructed from normal hematopoiesis and immunity, thereby achieving a therapeutic treatment purpose.
  • hematopoietic stem cell transplantation 2) Perform immunological recognition of hematopoietic stem cell transplantation, and / or carry out "transplantation of tissues, organs, cells, and factors related to autoantigen immune recognition". If partial myeloablative or non-myeloablative methods are used, hematopoietic stem cell transplantation can also be performed without immunological recognition, and the original immune recognition hematopoietic stem cells can be directly used for secondary self-antigen immune recognition.
  • Complement-dependent cytotoxicity method After the monoclonal antibody binds to the corresponding target cell, the complement is added, and the target cell is dissolved and destroyed by activating the complement pathway to achieve purification.
  • B cell monoclonal antibodies such as: CD9, CD10, CD19, CD20
  • T cell antibodies such as: CD2, CD3, CD4, CD8, etc.
  • AML-223 (anti-CD14 mAb) plus immune complement purification can be used to treat acute myeloid leukemia, which is similar to Alio-bone marrow transplantation.
  • Immunotoxin-mediated cytotoxicity method which combines specific antibodies with cytotoxin (ricin toxin) to form an immunotoxin, which has a powerful effect of killing specific cells and is specific.
  • recipients Patients receiving grafts or patients with immune system diseases, referred to in this specification: recipients. This recipient can be either a human or an animal. 10. Provide donor or graft antigen, referred to in this specification: donor. This donor can be either a human or an animal.
  • the early immunological recognition hematopoietic stem cells and/or late immune recognition hematopoietic stem cells of the present invention are performed twice in vitro, or in the donor, or in the recipient, on the donor's graft, and/or the variant autoantigen.
  • Immune recognition Because this autoantigen immune recognition differs from the immune recognition of the autoantigen by the immune system in the embryonic stage in which the human or animal naturally grows, it is an autoantigen including a donor's graft, and/or a variant self antigen.
  • the immunological recognition is to identify the graft and/or the self-antigen that causes mutation of the immune system disease as a normal autoantigen, and therefore, in the present specification, it is referred to as: secondary immune recognition.
  • secondary immune recognition is also referred to as: re-existing autoantigen immune recognition, or secondary self-antigen immune recognition, or self-antigen secondary immune recognition.
  • In the present invention from egg cells to obtaining early immune recognition hematopoietic stem cells and/or late immune recognition hematopoietic stem cells and/or grafts, to intermediate stages of obtaining thousands of cells and/or incubated grafts that have undergone immunological recognition,
  • In vitro culture tools such as cell culture flasks, culture dishes, and cultures are used to develop, culture, and self-antigen identification in in vitro culture tools such as culture flasks, culture dishes, and culture instruments.
  • test tube Abbreviation in the manual: test tube.
  • An immunomagnetic bead separation method can be performed using a specific antibody of a certain cell.
  • a cell suspension containing a certain tissue of a particular cell can be used.
  • tissue of a particular cell can be used.
  • Isolation of cells such as dendritic cells (DC):
  • DC dendritic cells
  • Various methods can be used, which can include the following protocols: The thymus tissue is ground into a single cell suspension by a single cell grinder, containing 6 X 10 8 PBMC suspension 9 ml containing 1 sleep ol / L EDTA calcium-free magnesium PBS), add anti-CD32 monoclonal antibody blocking solution 0. 4ml, 37 °C 5% C02 culture for 10min, then add StemSepTM human DC enriched mixed antibody magnetic beads lml, fully mixed, 37 Incubate at 5% C02 for 30 min, pass through a sterile magnetic sieve column, collect unlabeled cells of interest, and wash 3 times with PBS.
  • IX 105 cells were taken, and the cell phenotype was analyzed by fluorescent antibody labeling and flow cytometry, and the remaining cells were cultured in an IMDM medium containing 10 ng/ml of GM-CSF and 10 ng/ml of IL-4 in 10% FCS.
  • NK cells 9 ml of 4 ⁇ 108PBMC calcium-free magnesium PBS suspension was isolated and assayed in the same way as DC, and the remaining cells were cultured in IMDM medium containing 10% FCS.
  • T cells 9 ml of 4 ⁇ 108PBMC calcium-free magnesium PBS suspension was isolated and detected by DC separation. The remaining cells were cultured in IMDM medium containing 10% FCS.
  • pancreatic stem cells isolation and culture of pancreatic stem cells: (Wen Jin, Zhang Yuhai et al. Journal of Jiangxi Medical College, 2002, Volume 42, Issue 3)
  • the pancreas was added to 4 CHank's solution and cut into tissue pieces of 1 mm in diameter. After repeated washing, 0.5 rag/ml of collagenase was added, and the solution was shaken at 37 ° C for 15 minutes. The reaction was stopped by adding 4 CHank's solution. Centrifuge three times at 800 rpm, and add the supernatant to 4 CHank's solution without calf serum. Pass through a 150 mesh sieve and add 4 CHank's solution containing 10% calf serum to the resulting cell pellet.
  • the cell suspension was added to a culture solution of 0.1 mg/ml ConA to remove non-islet cells such as fibroblasts, and 5% C0 2 37 ° C pzyh 24 hours later, 5 ⁇ 10 5 /piece was added.
  • 20 ng/ml BFGF (basic fibroblast growth factor) 11. 1 mmpl/1 glucose 10% calf serum culture flask was continued to be cultured, and after 7-10 days, EDTA was diluted with 0. 05% trypsin, digested and passaged. cell. After five weeks, the culture medium was changed to low-sugar (2.5 mmol/1) 4 CHank's solution containing 2% calf serum, and BFGF was removed. 10 mM nicotinamide was added to induce cell differentiation to form islet cell-like cell colonies. The cells therein are islet stem cells.
  • Treatment a Knock out the self-antigen immune recognition library or the cells that store the self-antigen recognition library. And/or after knocking out these cells, the same type of cells into which the autoantigen immune recognition library is stored are not included, including cells such as stem cells and/or progenitor cells that can be differentiated into these cells, and stored for self-antigen immune recognition. After the new autoantigen immune recognition library.
  • Immunological cells such as T cells and B cells that have been negatively selected and positively selected in the thymus and bone marrow in peripheral blood and immune tissues.
  • Treatment a. Knock out immune cells such as T lymphocytes, B cells, etc. in the thymus, bone marrow, peripheral blood, and immune tissues.
  • immune cells such as T cells and B cells that have been negatively selected and positively selected in the thymus and bone marrow after re-immunization with autoantigen are recognized.
  • Self-antigen recognition factors as well as self-antigen recognition factors, and pro-antigen recognition factors, which may be factors or cells. Medical practice indicates that it may be present in tissues, organs, cells, and factors related to immune recognition of autoantigens such as thymus and/or bone marrow during the period of self-antigen immune recognition.
  • Treatment a. Look for cells that secrete these factors, do cell culture and input into the body for treatment.
  • organ tissues containing cells or secretory factor cells such as thymus and/or bone marrow, and/or intooculation of these organ tissues into a single cell suspension.
  • Thymus and/or bone marrow cells undergoing self-antigen immune recognition are also known as Thymus and/or bone marrow cells undergoing self-antigen immune recognition.
  • Treatment a. Use the original thymus and bone marrow cells in the body.
  • Treatment a. Input early non-immune recognition of hematopoietic stem cells, thymic T progenitor cells and / or B progenitor cells in the bone marrow.
  • Treatment a. Enter early non-immune recognition of hematopoietic stem cells.
  • the present invention can be applied not only to humans but also to animals, but in the description of the present invention, it is mostly represented by humans, including various embodiments in the specification. However, the present specification and various embodiments can also be applied to various animals. Organ transplantation can be performed between humans or animals of the same species, or organ transplantation can be performed between humans or animals of different species. Or transplanting between the immune systems of different species of humans or animals, using the animals of the transplanted immune system for immune system diseases or organ transplant treatment trials. Not only can you carry out various animal experiments, but you can also Organs are transplanted into animals, and the organs of the animals grow on the animals. When they grow up, they can be transplanted to the human body, and then transplanted from the animals to the human body and other diseases.
  • the fetal non-immune recognition stem cells perform an immune recognition on the autoantigens of various tissues and organs in the body, and store the results of this recognition into the autoantigen. Lifetime memory in the immune recognition library. After the immune system compares various antigens entering the body or antigens that are self-mutating, this memory is compared with the memory of the identification, and the antigens that are different from the autoantigens of various tissues and organs in the body are found. Generate an immune attack.
  • the transplantation of various organs in the body includes: liver transplantation, kidney transplantation, islet transplantation of diabetes, etc., because the consistent organs and / or tissues are foreign antigens, and the autoantigens in the self-antigen recognition memory in the body are different, so Inevitably, a rejection reaction occurs.
  • HLA matching is required before surgery, preferably all matches, at least semi-match is required, so that the antigen of the graft or graft antigen is as much as possible in the memory of the memory recognized by the autoantigen in the body, and at the same time After surgery, it is necessary to take anti-rejection drugs for life, such as cyclosporin (CSA), etc., taking anti-rejection drugs, it costs about 5,000 yuan a month, and the long-term cost is very expensive.
  • CSA cyclosporin
  • immune system diseases such as: systemic lupus erythematosus, rheumatism, rheumatoid arthritis, glomerulonephritis, scleroderma, etc., including immune diseases caused by self-antigenic changes caused by various causes of diseases, For example, cirrhosis caused by viral hepatitis is an immune attack caused by the immune system's own immune recognition error, causing physical damage.
  • the invention can alleviate or cure the inevitable rejection reaction of transplantation of various organs, so that organ transplantation and tissue cell transplantation can be used to treat many diseases which are currently untreated, such as: Diabetes can be treated by islet cell transplantation. It can also alleviate or cure various immune system diseases (AIDs) such as systemic lupus erythematosus, etc. These diseases are diseases in which the immune system causes immune recognition errors in its own tissues and causes physical damage. It can also treat a variety of other diseases that can be treated by this method, such as: congenital genetic diseases such as hematopoietic diseases.
  • AIDs immune system diseases
  • Zhagani's mesenchymal cells extracted from human bone marrow, are injected into the fetus of the sheep's uterus, and human cells grow into part of the sheep's heart, skin, muscles, fat and other tissues, but the number of human cells at that time very few.
  • the team he led has made further progress on the basis of the original, that is, surprisingly greatly increased the content of human cells in some sheep organs. According to the demonstration at a meeting in early December, the human cell content in the liver of the tested sheep has reached 7-15%.
  • the measurement results show that the MHC content in the undivided embryonic stem cells is low, but stable. Embryonic stem cells, the embryos, have not yet differentiated into "raw" cells of various tissues. According to early human embryo research, scientists have suggested that embryonic stem cells may not be discovered by the body's mechanism of defense against foreign cells. Hugh O'Kinkos of Harvard Medical School in the United States said that the new results prove that embryonic stem cells cannot escape the detection of the immune system.
  • HLA tissue-associated antigen
  • the mouse does not have any immune function and is completely receptive to the graft. It is completely acceptable. While the present invention uses a mouse having NK cell function (having a certain immune function), the successful transplantation of the test examples of the present invention proves that the present invention does overcome the rejection reaction, and proves the theory and the theory of the present invention. The correctness of practice.
  • self-antigen immune recognition library (although it is not yet fully known about the specific storage location of this "self-antigen immune recognition library", in which cell, in which organelle of the cell, and the specific location, but at least It is known that some cells harboring autoantigen immune recognition libraries, such as: cortical epithelial cells and medullary dendritic cells associated with immune recognition in the thymus, and/or corresponding cells associated with immune recognition of bone marrow cells (see “ Cellular and Molecular Immunology” Chapter 6, Section Chapters 7 and 9), this "self-antigen immune recognition library” determines the self-antigen reference standard for immunological recognition after birth.
  • the autoimmune disease is caused by self-antigen mutation
  • the allogeneic organ or the mutated tissue antigen does not enter the fetus when the fetus performs autoantigen immunological recognition
  • the "self antigen" established after autoantigen immune recognition
  • the antigenic gene of this transplanted organ is not recognized as its own antigen, so it will be issued. Give birth to an immune response.
  • Taiwan Regeneration Biotechnology Co., Ltd. has developed "Umbilical Cord Blood Stem Cell-Free Medium", which enables umbilical cord blood stem cells to increase value by 30 times within five to seven days, increase to seven thousand times within five weeks, and maintain differentiation. The characteristics and potential of stem cells.
  • a method for obtaining and applying stem cells for treating diseases of the immune system and rejection is provided, characterized in that the self-antigen is used for treating diseases of the immune system and rejection diseases.
  • the self-antigen is used for treating diseases of the immune system and rejection diseases.
  • Early immune recognition of secondary immune recognition The source of hematopoietic stem cells and/or late immune recognition hematopoietic stem cells is obtained from the following pathways:
  • a clonal cell formed from the graft recipient or the somatic cell nucleus of the immune system and the diseased patient to the human egg from which the nucleus is removed;
  • clonal cells formed from the recipient's somatic cell nucleus to the animal's egg from which the nucleus is removed; c.
  • the fertilized egg is formed from the sperm or egg of the recipient and the donor or other human or animal sperm or egg.
  • including early immune recognition hematopoietic stem cells and/or late immune recognition hematopoietic stem cells are expanded in vitro or transplanted into an intermediary for amplification.
  • Including early immune recognition of hematopoietic stem cells and/or late immune recognition of hematopoietic stem cells for secondary immune recognition is carried out in an in vitro test tube, either in an intermediary or in a recipient.
  • the early immune recognition hematopoietic stem cells and/or the late immune recognition hematopoietic stem cells are subjected to secondary immune recognition in the body.
  • transplantation using myeloablative, partial clear pulp or non-myeloablative, transplanting stem cells that have undergone immunological recognition by graft or graft antigen secondary immune recognition in recipients to recipients Then transplant the graft into the recipient.
  • Blood stem cell bone marrow transplanted animals Transplantation without immunological recognition of stem cells for self-antigen recognition: Animals are made an animal with an immune system recognized by humans or affected antigens. In particular: including transplantation of immune organs into the intermediary: in particular: including input initiation factors, and/or termination factors.
  • the principle of the present invention is based on a research team led by Esmai l Zanjani of the University of Nevada, USA, which demonstrates that human organs can be planted in intermediaries. And is recognized by the animal's immune system as its own tissue, self-antigen, without rejection.
  • the T cells in the thymus pass through the thymocyte precursor (prothymocyte) in the early thymus as CD4XD8-double-negative cells, which then develop into CD4 + C8 + TCR + double positive cells, which pass through the MHC class I with the surface of the thymic cortex epithelial cells or A positive selection for binding of MHC class II molecules/self peptides, and dendritic cells that express high levels of MHC class I or MHO class II molecules and autoantigens at the junction of the thymic cortex and medulla with cell surface ( DC) and macrophage ( ⁇ ) for negative selection of high affinity binding, through self tolerance or clonal deletion, become CD4+CD8 that can recognize both foreign antigens and non-antigens.
  • thymocyte precursor prothymocyte
  • CD4XD8-double-negative cells which then develop into CD4 + C8 + TCR + double positive cells, which pass through the MHC class I with the surface of the thy
  • CD4-CD8+ single positive sputum cells CD4-CD8+ single positive sputum cells.
  • progenitor B cells undergo developmental stages of early progenitor B cells, late progenitor B cells, pre-B cells, and immature B cells in the bone marrow, and are cloned and deleted to develop autoimmune-tolerant mature B cells ( Chapter IX B lymphocytes).
  • thymus and/or bone marrow organs in the period of self-antigen immune recognition, and/or certain tissue cells in these organs.
  • thymocytes, bone marrow cells, early immune recognition, hematopoietic stem cells, and/or advanced immune recognition of hematopoietic stem cells and/or use their own late-stage immune recognition to identify hematopoietic stem cells, re-existing autoantigen immune recognition, and mutating autoantigens and / or the antigen of the transplanted tissue organ is recognized as an autoantigen, and a new "autoantigen immune recognition library" including a self-antigen recognizing the graft antigen or mutation is established, thereby avoiding a rejection reaction or an immune reaction.
  • the original "autoantigen immune recognition library" in the body is not deleted, but the tissues and organs and cytokines related to the immune recognition of the autoantigen are input into the body, including: "autoantigen immune recognition factor", and/or secretion.
  • the organ of the "autoantigen immune recognition factor”, or all cells of the tissue or organ, or certain types of cells secreting "autoantigen immune recognition factor", and/or secretory "promoting" autoantigen immune recognition factor The secretion of cells such as cells promotes the identification of hematopoietic stem cells by cells that carry out their own antigenic immune recognition factors, or directly inputs these factors.
  • thymus and/or bone marrow organs in the period of self-antigen immune recognition, and/or certain tissue cells in these organs. And/or thymocytes, bone marrow cells, early immune recognition, hematopoietic stem cells, and/or advanced immune recognition of hematopoietic stem cells, and/or use their own late-stage immune recognition to identify hematopoietic stem cells, re-existing autoantigen immune recognition, allowing the body to re-identify the transplant
  • the antigen or the mutated autoantigen, and the re-identification of the graft antigen or the variant autoantigen is added to the original "autoantigen immune recognition library", so that the newly generated one can recognize the foreign antigen and not react with the autoantigen.
  • CD4+CD8- or CD4 D8+ single positive T cells and B cells The immune system does not immunoreact with the graft antigen or the variant autoantigen.
  • the monoclonal antibody binds to the corresponding target cell and then adds complement, and the target cell is lysed and broken by activating the classical pathway of complement to achieve purification.
  • Immunotoxin-mediated cytotoxicity method which combines specific antibodies with cytotoxin (ricin toxin) to form an immunotoxin, which has a strong tumor killing effect and specificity.
  • Coating microspheres or metal particles with a monoclonal antibody labeling the target cells to increase the density of the cells or generating magnetic properties, and then removing the target cells by precipitation or magnetic field adsorption, and performing anti-dendritic cell demyelination treatment of the cells, or Other monoclonal antibodies that complement the cells of the autoantigen immune recognition library, combined with complement, and/or antibody binding to cytotoxic drugs, perform myeloablative treatment of these cells.
  • multiple B-cell monoclonal antibodies CD9, CD10, CD19, CD20
  • T-cell antibodies CD2, CD3, CD4, CD5, etc.
  • cytotoxic drugs such as castor bean toxin
  • the target cells are labeled to increase the density or magnetic properties, and then the cells are subjected to myeloablative treatment by precipitation or magnetic field adsorption.
  • the present invention is a cell for stem cells, progenitor cells, etc., which is immunologically recognized and/or immunologically associated with autoantigens, which have not yet been and/or have been immunologically recognized for autoantigens of various tissues and organs in the body, for example, from fertilized eggs.
  • Stem cells associated with autoantigen immunological recognition and/or immunity such as hematopoietic stem cells, and/or various types of immune-related hematopoietic progenitor cells, are produced at various developmental stages of embryos of gestational age below 7 months of age.
  • the original, mature, hematopoietic stem cells with immune recognition function are killed, scored or killed by the bone marrow transplantation of the marrow, part of the marrow, or the unclear marrow.
  • organs, organs, cells, and factors related to immune recognition of autoantigens include: organs such as thymus, spleen, lymph nodes, and bone marrow, and various cells, hormones, and various cells involved in the immune recognition of self antigens contained in these organs. Cytokines, peptides, proteins, tissues, tissue fluids, cells, and more.
  • the immunological recognition of hematopoietic stem cells required for re-immunization of autoantigens is obtained from the following pathways:
  • cloning cells or fertilized eggs of different origins are developed into a non-immune-recognized stem cell stage embryo in the intermediary, and the immune-free stem cells are extracted from the embryo;
  • the immune recognition hematopoietic cells are subjected to the period of self-antigen immune recognition, and the latent immune recognition hematopoietic stem cells extracted from the stillbirth are aborted;
  • clonal cells or fertilized eggs of different origins are developed into an early immune recognition hematopoietic stem cell and/or an embryo of the late immune recognition hematopoietic stem cell stage, and early immune recognition hematopoietic stem cells and/or late immune recognition hematopoiesis are extracted from the embryo.
  • Stem cells, or cultured in vitro differentiate into early immune recognition hematopoietic stem cells and/or late immune recognition hematopoietic stem cells.
  • the extracted early immune recognition hematopoietic stem cells and/or late immune recognition hematopoietic stem cells can be used not only to treat immune system diseases, but also to treat a variety of other diseases, such as: hematopoietic diseases caused by congenital inheritance, and the like.
  • Amplification is carried out in a test tube.
  • Early immune recognition hematopoietic stem cells and/or late immune recognition hematopoietic stem cells obtained from several pathways in a. in vitro test tubes, b. mediators, cultured, expanded, expanded to a suitable number for transplantation in vivo , performing immunological recognition of the donor's graft antigen and/or the variant autologous antigen in the recipient. Whether amplification is needed can determine the number of hematopoietic stem cells and/or late immune recognition hematopoietic stem cells according to the required early immune identification, such as: The amount required for secondary immune recognition in the transplanted mediator has been met, perhaps No amplification is required.
  • the early immune recognition hematopoietic stem cells and/or the latent immune recognition hematopoietic stem cells are transplanted into the intermediary, and the intermediary used may be: an immunodeficient animal or an immunodeficient animal.
  • the early immune recognition hematopoietic stem cells and/or the late immune recognition hematopoietic stem cells are transplanted into the intermediary to identify the secondary immune recognition.
  • the graft or graft antigen and/or the variant internal antigen in the recipient is transferred to the intermediary.
  • Transplant recipients and/or donor grafts or graft antigens and/or variants of autologous antigens in recipients into vitro in vitro may be It is a tissue, a cell, or an organ, etc.
  • test tube which may be an organ It can also organize, cells, factors, and more.
  • Transplantation of graft or graft antigen into recipients requires whether the transplant can be determined according to the condition to be treated. For example, treatment of immune system disease does not require transplantation.
  • the original self-antigen identification library is completely destroyed, that is, using bone marrow transplantation methods, such as: using radiation, and / or using cytotoxic drugs, and / or using monoclonal antibodies combined with cytotoxic drugs will be the original All cells in the body that store their own antigen recognition library are killed.
  • the primary immune recognition hematopoietic stem cells and/or the late immune recognition hematopoietic stem cells, and/or the "organisms, organs, cells, and factors associated with autoantigen immune recognition” are subjected to secondary immune recognition in the recipient. Or input through the recipient Stem cells that have been subjected to secondary immunological recognition in vitro and/or cells that store new "autoantigen immune recognition library” cells, and add new library contents.
  • the specific method used, the results of the experiment can be selected according to the specific needs of the disease.
  • mice with thymic function loss have no immune function of T lymphocytes, such as BALB/ cA-nude nude mice, CW Friis of Denmark in 1973 found spontaneously mutated hairless mice in BALB/cA inbred mice.
  • the mutant mouse has thymic dysplasia, immune T cell loss, and has been cultured as BALB. /cA-nude, while C.
  • B-17 SCID mice have no mature B and T lymphocytes, so the thymus, spleen, and lymph nodes
  • the weight is less than 30% of normal, and histologically manifested as a significant defect in lymphocytes.
  • the thymus is surrounded by multiple adipose tissue, no cortical structure, only the remaining medulla, mainly composed of epithelial-like synthetic fibroblasts, and occasionally focal follicular lymphocytes.
  • the spleen white pulp is not obvious, the red pulp is normal, and the spleen has no lymphocyte aggregation, mainly composed of reticular cells. There is no obvious cortical area in the lymph nodes, the cortical area is missing, and there are reticulocytes.
  • Submucosal and bronchial lymph nodes are rare in the small intestine, and there is no lymphoid accumulation in the structure. Both thymus and spleen function are absent and have neither T lymphocyte function nor B lymphocyte function. There were no mature B and T lymphocytes in N0D-SCID mice.
  • multiple organs in tissues, organs, cells, and factors related to immune recognition of autoantigens are important organs for early immune recognition of hematopoietic stem cells and/or late immune recognition of hematopoietic stem cells for immune recognition, in which multiple organs are In blood, tissue fluids, various cells, or entire organs, there should be activating recognition factors that promote, promote, and/or halt early immune recognition of hematopoietic stem cells and/or late immune recognition of hematopoietic stem cells for self-antigen recognition, motivating recognition factors, and The recognition factors are terminated, which can be: polypeptides, proteins, tissues, cells, organs, and the like.
  • cells associated with immune recognition cells in various organs, which are in the initial stage of embryonic autoantigen immune recognition and/or in the recognition phase and/or termination phase, may be present in the blood and
  • the composition of the tissue fluid, etc. is added to the culture medium, and/or imported into the body of the human or intermediary to initiate early identification of hematopoietic stem cells and/or late immune recognition in a test tube or in a human or intermediary body.
  • Hematopoietic cells are recognized for their own antigens.
  • hematopoietic stem cells and/or latent immune recognition hematopoietic stem cells By identifying hematopoietic stem cells and/or latent immune recognition hematopoietic stem cells, early immune recognition hematopoietic stem cells and/or late immune recognition hematopoietic stem cells can be extracted from these organs. If not, early immune recognition hematopoiesis can be directly extracted from the corpse of the fetus. Stem cells and/or late immune recognition of hematopoietic stem cells.
  • organs, organs, cells, and factors related to immune recognition of autoantigens include: organs such as thymus, spleen, lymph nodes, and bone marrow.
  • the non-immune recognition stem cells of these fetuses are not immunologically recognized by the autoantigens of various tissues and organs in the fetal body. (It can also be obtained from various other sources, such as other kinds of stem cells, such as single cells such as adipose stem cells.
  • the bone marrow transplantation method is used to remove immune-related cells, including "autoantigen immune recognition library” and cells storing an autoantigen immune recognition library.
  • drugs such as: cyclophosphamide, and/or radiation irradiation, such as : cobalt 6 °, medical linear accelerators, high-energy X-ray whole body irradiation
  • the human body's original method has been autoantigen immune recognition of hematopoietic stem cells and / or hematopoietic progenitor cells in vivo hematopoietic stem cells to kill the original, the original use of drugs in vivo T lymphocytes Immune cells are killed, for example: Cytotoxic drugs formed by binding CD3 monoclonal antibodies to cytotoxins plus complement input to kill T lymphocytes in the body, etc., killing immune cells in the body's immune system, and / or "self antigen immune recognition library" and drugs that store cells with their own antigen immune recognition library.
  • the termination factor may be one or more polypeptides, one or more proteins, or one or more tissue cells, or a Or multiple tissues and organs.
  • Simultaneous input (other cells can be included in the input cells, and the original hematopoietic stem cells in the body can be killed without using drugs and/or radiation according to different diseases and different case needs. No immune recognition of stem cells entered into the body).
  • hormones autoantigen-immuno-recognition factors
  • promoting autoantigen-immuno-recognition factors
  • promoting in the body "immunity-recognition-associated tissues, organs, cells, and factors”
  • the input amplified non-immune recognition cells will immediately perform a new immune recognition.
  • transplanted organs and the self-organizing tissue that produces the mutation as its normal tissue and organ, which greatly reduces the intensity of the immune attack, and does not even generate new immune attacks. Thereby treating the disease, it can greatly reduce the medical expenses after transplantation of various organs.
  • various immune system diseases can be cured or alleviated, and since the transplanted organs, tissues or cells can be re-identified, various diseases such as: islet transplantation for islet transplantation, and the like can be treated by organ or tissue transplantation.
  • the input is a healthy fetal amplified non-immune recognition stem cell, it can produce normal blood cells, thus curing various diseases of the hematopoietic system caused by congenital inheritance such as: congenital chain-like erythrocyte anemia. It is also possible to treat other types of diseases by this method as long as the treatment is beneficial to the recovery of the patient.
  • the foreign tissue entering the human body or the self-organizing tissue that produces the mutation is regarded as a foreign body, which generates an immune attack and therefore does not occur at present.
  • the use of immunosuppressive agents in the treatment of the problem of reduced immune function is regarded as a foreign body, which generates an immune attack and therefore does not occur at present.
  • the extraction of stem cells from a fragmented stillbirth has no moral problems, and the diversity of HLA sources of fetal immune-related stem cells, such as hematopoietic stem cells, can be guaranteed. Therefore, the hematopoietic stem cells extracted from the stillbirth obtained by abortion curettage are very suitable for the treatment of adult autoantigen re-identification and immune system diseases.
  • the non-immune-recognizing stem cells used in the present invention such stem cells: stem cells derived from abortion of gestational fetuses having a gestational age of less than three months, which are immunologically recognized and/or immune-related, such as hematopoietic stem cells and/or Various types of immune-related hematopoietic progenitor cells, raw embryonic stem cells obtained from various sources in vivo or in vitro, and natural breeding or artificial culture or artificial intervention in embryonic bodies or culture flasks, and original embryos Stem cells or various stem cells obtained from various sources differentiate or transform into various stages of self-antigen immune recognition and/or immune-related stem cells, which may include: totipotent stem cells (embryonic stem cells), pluripotent stem cells ( For example, hematopoietic stem cells and/or various types of stem cells, such as hematopoietic stem cells, and/or various types of stem cells, such as hematopoietic stem cells, and/or immune-related
  • Embryonic stem cells that is, "primitive" cells (ie, pluripotent stem cells) that have not yet differentiated into various tissues in the embryo can differentiate into all cells possessed by the adult body. More than 200 kinds of cells in the human body can be produced by reproduction. Finally, the cells can produce tissues, tissues and synthetic organs, and become human bodies. Embryonic stem cells are mainly found in early embryos. There is a process in embryonic development. The combination of sperm and egg produces fertilized eggs. The fertilized eggs are further divided into morula.
  • the morula is further divided into blastocysts.
  • the blastocysts produce inner cells. At this stage, only one is extracted. Cells can develop into adult bodies, so they are omnipotent stem cells. Multi-functional The cell has been developed late.
  • stem cells in the bone marrow that are immunologically recognized and/or immune to autoantigens, such as hematopoietic cells, which are pluripotent stem cells.
  • Hematopoietic stem cells are tissue-specific stem cells derived from the mesoderm cells of the embryonic yolk sac and successively migrated to the hematopoietic organs, liver, spleen and bone marrow in the embryo, maintaining the self-determination by asymmetrical mitosis.
  • HSC Hematopoietic cells
  • progenitor cells Can differentiate into a wide variety of progenitor cells. It can be differentiated into white blood cells, red blood cells, platelets, etc. in the bone marrow, which is pluripotent. The other is unipotent or multi-functional, can only differentiate into a kind of cell, or differentiate into two closely related cells, called unipotent stem cells (such as multiple hematopoietic progenitors). As long as the various stages and types of stem cells can be induced to differentiate into hematopoietic stem cells by various stem cell lines and transformed into non-immune-recognized stem cells, they can be used for the re-identification of adult autoantigens and the treatment of immune system diseases.
  • Use drugs that allow stem cells in quiescent phase to enter the active phase such as: 5-FU, which inhibits hematopoietic stem cell division by inhibiting DNA cleavage, thus stimulating the body to let the hematopoietic stem cells in quiescent phase enter the active phase, and then proceed Radiation exposure or drugs kill active hematopoietic stem cells; various hematopoietic stimulating factors can also be used to stimulate the quiescent hematopoietic stem cells in the body to enter the active phase, and then irradiate or kill the active hematopoiesis stem cell.
  • 5-FU which inhibits hematopoietic stem cell division by inhibiting DNA cleavage
  • Monoclonal antibody fragments such as: monoclonal antibody 3A5 Fab' fragment, bispecific antibody, trispecific antibody, antibody cytokine fusion protein, and the like.
  • Novel genetically engineered antibodies continue to emerge, such as humanized antibodies, monovalent small molecule antibodies (Fab, single-chain antibodies, single-domain antibodies, hypervariable region polypeptides, etc.), multivalent small molecule antibodies (double-chain antibodies, triple-chain antibodies, mini Antibodies, etc., certain types of antibodies (bispecific antibodies, trispecific antibodies, antibody cytokine fusion proteins, antigenized antibodies, intracellular antibodies, catalytic antibodies, immunoliposomes, etc.) and antibody fusion proteins (immunotoxin, immunity) Adhesin) A combination of various cytotoxins (such as ricin, etc.).
  • Fab single-chain antibodies, single-domain antibodies, hypervariable region polypeptides, etc.
  • multivalent small molecule antibodies double-chain antibodies, triple-chain antibodies, mini Antibodies, etc.
  • certain types of antibodies bispecific antibodies, trispecific antibodies, antibody cytokine fusion proteins, antigenized antibodies, intracellular antibodies, catalytic antibodies, immunoliposomes, etc.
  • An immunoconjugate comprising a chemically cross-linked monoclonal antibody can be used to produce hematopoietic stem cells, and/or various immune progenitor cells, and/or various immune cells.
  • Antibody-directed chemotherapy antibody- Mediated chemotherapy
  • a cytotoxic chemical such as arsenic, cisplatin, fluorouracil, vincristine , doxorubicin, etc.
  • Immunotoxin therapy the toxin is linked to a monoclonal antibody, and the prepared immunotoxin is a stem cell that has been immunologically recognized in the recipient, or one of them, Several cells have specific strong killing activity.
  • toxins There are two types of commonly used toxins: one is phytotoxin, including castor bean toxin (RT), acacia toxin (abrin), and bitter melon toxin (MD).
  • RT castor bean toxin
  • abrin acacia toxin
  • MD bitter melon toxin
  • cytotoxins including diphtheria toxin (DT), Pseudomonas aeruginosa exotoxin (PE), and the like.
  • hematopoietic stem cells and/or various immune progenitor cells can be used, such as monoclonal antibodies such as CD34+ or A133, such as hematopoietic stem cell-specific antibodies, combined with cytotoxic drugs, such as cytotoxic drugs such as ricin and diphtheria toxin.
  • Fig. 1 is a flow chart showing a method for extracting, culturing and amplifying hematopoietic stem cells and/or late immune recognition hematopoietic stem cells of the present invention
  • Figure 2 is a flow chart showing the treatment of adult autoantigen re-identification and immune system diseases by organ transplant patients and patients with immune system diseases of the present invention
  • Figure 3 Is a method for obtaining, culturing, amplifying, and performing secondary immune recognition of a hematopoietic stem cell and/or a late immune recognition hematopoietic stem cell in vitro, in vitro, for treating a graft rejection reaction, and/or Immunology Flow chart of the systemic disease method;
  • Figure 4 is a diagram of the present invention for early immune recognition of hematopoietic stem cells and/or advanced immune recognition of hematopoietic stem cells in vitro, extraction, expansion, and secondary immune recognition in recipients to treat graft rejection, And/or a flow chart of the immune system disease method;
  • Figure 5 is a flow chart showing an experimental method for performing secondary immune recognition by an early immune recognition hematopoietic stem cell and/or a late immune recognition hematopoietic stem cell in a mediator;
  • Figure 6 is a flow chart showing another method for extracting, culturing and amplifying an early immune recognition hematopoietic stem cell and/or a late immune recognition hematopoietic stem cell for bone marrow transplantation of the present invention
  • Figure 7 is a flow chart showing the treatment of adult autoantigen re-identification and immune system diseases in another organ transplant patient and immune system disease patient of the present invention.
  • Figure 8 is a flow chart showing the extraction of hematopoietic stem cells and thymocytes and bone marrow cells of one embodiment of the present invention
  • Figure 9 is an illustration of an autoantigen re-identification and immune system disease of an organ transplant in an animal of the present invention.
  • Fig. 1 A flow chart of a method for extracting, culturing and amplifying hematopoietic stem cells and/or late immune recognition hematopoietic stem cells according to the present invention. Stem cells without immune recognition are obtained, and the following is an example of the acquisition of hematopoietic stem cells.
  • the stillbirth from abortion The liver and bone marrow of the stillbirth are peeled off under the microscope. Because the bone marrow is relatively easy to obtain, it is best to obtain stem cells without immune recognition from the bone marrow, but because the fetus is liver hematopoietic, it can also be Stem cells that are not immune-recognized are obtained in the liver, or stem cells that are not immune-recognized from other organs or tissues.
  • Immunomagnetic beads-flow cytometry two-step sorting method can be used to sort out non-immunologically recognized stem cells from bone marrow, for example, monoclonal antibodies and packages using stem cell antigen (Stemcellantigen, Sea) specifically expressed on the surface of stem cells.
  • stem cell antigen Stemcellantigen, Sea
  • Hematopoietic stem cells which are not immunologically recognized in the bone marrow are separated by a magnetic adsorption cell sorting method (MACS) by a second antibody on the surface of the magnetic particle.
  • MCS magnetic adsorption cell sorting method
  • Other methods can also be used to sort out stem cells that are not immunologically recognized.
  • the sorted non-immune-recognized stem cells are cultured in a culture medium containing a combination of different hematopoietic growth factors, and may be a serum-free medium.
  • the currently identified stimulatory and inhibitory factors that act on the expansion (proliferation) of hematopoietic stem cells are quite numerous, but only a few cytokines can function alone.
  • a variety of hematopoietic growth factors can be added to serum-free medium, such as SCF, IL-3, IL-6, TP0, Flt3-L, G-CSF, and Epo.
  • SCF, IL-3 and GM-CSF mainly maintain the survival of the original hematopoietic cells, but can not induce their proliferation.
  • the proliferation of the cells is required to induce proliferation.
  • the bone marrow mesenchymal stem cells can be co-cultured with the hematopoietic stem cells. Hematopoietic stem cells have enhanced amplification. The number of nucleated cells (NC), surface antigens, and colony forming ability were monitored. The in vitro expansion culture is generally carried out for about 2 weeks. Of course, you can increase or decrease the number of days according to the actual situation.
  • Figure 2 is a flow chart showing the treatment of adult autoantigen re-identification and immune system diseases in patients with organ transplantation and patients with immune system diseases according to the present invention.
  • Non-immune-recognized stem cell bone marrow transplantation is prepared after transplantation of the host patient or in the case of a patient such as an immune system disease.
  • drugs that promote hematopoietic stem cell homing may be used according to the condition (hematopoietic stem cell (HSC) homing refers to HSC passing through the peripheral blood circulation into the recipient after intravenous transplantation) Recognition and localization in the bone marrow mediated by complex intermolecular interactions. And in vivo stem cell expansion treatment (exvi V0 ).
  • the stem cell factor SCF can be used in combination with G-CSF to stimulate expansion of the chimeric/progenitor cells (PBPC) in vivo.
  • the immune recognition of autoantigens is the time required for those factors to promote hematopoietic stem cells for autoimmune recognition and immune system reconstruction. At this time, it is necessary to track various immunological test indicators.
  • the organ transplantation stage and bone marrow transplantation, and the secondary autoantigen immune recognition stage can be performed simultaneously, or the graft antigen can be transplanted first for bone marrow transplantation, secondary self antigen immune recognition, and the body's immune system recognizes the graft antigen as After the autoantigen, organ transplantation is performed, which can increase the survival rate of the graft.
  • Example 3
  • Figure 3 It is a method for obtaining, culturing, amplifying and performing secondary immune recognition of a hematopoietic stem cell and/or a late immune recognition hematopoietic stem cell in vitro for human immunodiagnosis in vitro, and treating a graft rejection reaction, and/or Flow chart of the method of immune system disease.
  • Fertilized eggs (which can also be cloned egg cells) can be obtained in several ways: 1) After normal pregnancy, a miscarriage occurs, and no early immune recognition hematopoietic stem cells and/or late immune recognition hematopoietic stem cells, and/or transplants to be transplanted are extracted from stillborn fetuses. This graft may be organs, tissues, Cells, such as: liver, kidney, islet e cells, etc.
  • somatic cell nucleus of the recipient or donor is cloned into the egg cell of the same species as the recipient or donor, and the formed clonal egg cell develops in the test tube, or in the human body, or in the intermediary.
  • the somatic cell nuclei of the recipient or donor are cloned into the egg cell of the different species of the nucleus to form a cloned egg cell, which develops into an early immune recognition hematopoietic stem cell and/or late immunity in a test tube or in an intermediary or human body.
  • the hematopoietic stem cell stage is identified, and early immune recognition hematopoietic stem cells and/or late immune recognition hematopoietic stem cells, and/or grafts requiring transplantation are extracted.
  • the fertilized egg obtained by fertilization of the sperm or egg of the recipient and the sperm or egg of the same species, including the fertilized egg of the recipient and/or the donor parent and the mother egg, in a test tube or in a mediator or
  • the human body develops into an early immune recognition hematopoietic stem cell and/or a late immune recognition hematopoietic stem cell stage, extracting early immune recognition hematopoietic stem cells and/or late immune recognition hematopoietic stem cells, and/or transplants requiring transplantation.
  • Fertilized eggs can also be obtained from a variety of different pathways using different methods.
  • In vitro culture is carried out by culturing a fertilized egg in a culture flask containing a medium, adding a hematopoietic stem cell-directed differentiation factor, and differentiating into an early immune recognition hematopoietic stem cell and/or a late immune recognition hematopoietic stem cell.
  • the fertilized egg is implanted in an intermediary or a human body, such as a human uterus, or an animal such as a mouse or a rabbit, and the development time can be determined according to the specific needs of the medical condition, for example, waiting for embryo development.
  • a human body such as a human uterus, or an animal such as a mouse or a rabbit
  • the development time can be determined according to the specific needs of the medical condition, for example, waiting for embryo development.
  • the early immunological recognition hematopoietic stem cells and/or the late immune recognition hematopoietic stem cells are extracted by the set method, for example, using a flow cytometer. It is also possible to perform amplification in a culture flask after extracting early detection of hematopoietic stem cells and/or late immune recognition of hematopoi
  • the intermediary including 1. Transplantation of early immune recognition hematopoietic stem cells and/or late immune recognition hematopoietic stem cells into immunodeficient mediators, such as immunodeficient small BALB/cA-nude nude mice, SCID Mouse, NOD-SCID mice were performed in the body.
  • immunodeficient mediators such as immunodeficient small BALB/cA-nude nude mice, SCID Mouse, NOD-SCID mice were performed in the body.
  • immunodeficient mediators such as immunodeficient small BALB/cA-nude nude mice, SCID Mouse, NOD-SCID mice were performed in the body.
  • immunodeficient mediators such as immunodeficient small BALB/cA-nude nude mice, SCID Mouse, NOD-SCID mice were performed in the body.
  • Early immune recognition hematopoietic stem cells and/or late immune recognition hematopoietic stem cells and/or grafts can also be transplanted into the immunodeficient mediator body by
  • Non-immune recognition stem cells extracted from aborted fetuses, and/or isolated organs to be transplanted are extracted from aborted fetuses, and/or isolated organs to be transplanted.
  • transplanter or the intermediary is starting or undergoing immunization of the embryo at the stage of self-antigen immune recognition Identifying organs, and/or different types and types of tissues, cells, and/or internal organs associated with immune recognition that are selected for different types and types of organs that are transplanted according to design requirements, and/or different types of organs associated with immune recognition Contents such as tissue fluids are transferred to the intermediary. Deficiency of immune organ function in immunodeficiency mediators, and/or mediators undergoing radiation and/or drug therapy.
  • the initiation factor can be input according to the design requirements to enable early immune recognition hematopoietic stem cells and/or late immune recognition hematopoietic stem cells to perform autoantigen immune recognition.
  • an immunologically-recognized organ of an embryo or an intermediary who is starting or performing an embryo at the stage of self-antigen immune recognition and/or a different type, different type of cells selected from organs transplanted according to design requirements, and / or a starter factor, and/or a stop factor that already exists when the contents of different types, different types of tissue fluids inside the organ are in the intermediary.
  • Thousand cells that have been immunologically recognized are transplanted through the bone marrow and/or transplanted directly into the recipient.
  • Organs, tissues, and cells of the graft can be transplanted before, at the same time as, or after transplantation of the immunoreactive stem cells into the recipient. And / or treatment of immune system diseases.
  • the transplanter or the intermediary is starting or undergoing an immune recognition organ of the embryo at the stage of self-antigen immune recognition, and/or is selected according to the design requirements to transplant the embryo in the selected different types, different types of immunity with the autoantigen Identify relevant tissues, cells, and/or contents of different types, different types of tissue fluids inside the organs associated with the immune recognition into the test tubes.
  • Input activating factor to enable early immune recognition of hematopoietic stem cells and/or late immune recognition hematopoietic stem cells for autoantigen immune recognition.
  • an immunologically recognized organ of an embryo or an intermediary who is starting or undergoing an embryo at the stage of self-antigen immune recognition, and/or selected different types, types of cells in an organ transplanted according to design requirements, and / or contents of different types, different types of tissue fluid inside the organ into the test tube.
  • the abort factor can be input as needed to enable early immune recognition of hematopoietic stem cells and/or late immune recognition of hematopoietic stem cells to stop autoantigen immune recognition.
  • Organs, tissues, and cells of the graft can be transplanted after the stem cells that have been subjected to immunological recognition are transplanted into the recipient. And / or treatment of immune system diseases. Since the immune system in the body has already recognized the graft as a self-antigen, immune rejection does not occur even if the HLA of the graft does not match the recipient. At the same time, the intermediary can be used to cultivate the organ to be transplanted. The source of the organ that needs to be transplanted is further expanded.
  • Figure 4 is a diagram of the present invention for early immune recognition of hematopoietic stem cells and/or advanced immune recognition of hematopoietic stem cells in vitro, extraction, expansion, and secondary immune recognition in recipients to treat graft rejection, And/or a flow chart of the immune system disease method.
  • Fertilized eggs (which can also be cloned egg cells) can be obtained in several ways - 1) After a normal pregnancy, a miscarriage occurs, and no early immune recognition of hematopoietic stem cells and/or late immune recognition is extracted from stillborn fetuses. Hematopoietic stem cells, and/or grafts that require transplantation, this graft can be an organ, Tissues, cells, such as: liver, kidney, islet beta cells, etc.
  • somatic cell nucleus of the recipient or donor is cloned into the egg cell of the same species as the recipient or donor, and the formed clonal egg cell develops in the test tube, or in the human body, or in the intermediary.
  • the somatic cell nuclei of the recipient or donor are cloned into the egg cell of the different species of the nucleus to form a cloned egg cell, which develops into an early immune recognition hematopoietic stem cell and/or late immunity in a test tube or in an intermediary or human body.
  • the hematopoietic stem cell stage is identified, and early immune recognition hematopoietic stem cells and/or late immune recognition hematopoietic stem cells, and/or grafts requiring transplantation are extracted.
  • the fertilized egg obtained by fertilization of the sperm or egg of the recipient and the sperm or egg of the same species, including the fertilized egg of the recipient and/or the donor parent and the mother egg, in a test tube or in a mediator or
  • the human body develops into an early immune recognition hematopoietic stem cell and/or a late immune recognition hematopoietic stem cell stage, extracting early immune recognition hematopoietic stem cells and/or late immune recognition hematopoietic stem cells, and/or grafts requiring transplantation.
  • Fertilized eggs can also be obtained from a variety of different pathways using different methods.
  • Non-immune recognition stem cells extracted from aborted fetuses, and/or isolated organs to be transplanted are extracted from aborted fetuses, and/or isolated organs to be transplanted.
  • the transplanted embryo is beginning or undergoing an immunological recognition organ at the stage of autoantigen immune recognition to the recipient. And/or an input promoter that allows early immune recognition of hematopoietic stem cells and/or late immune recognition of hematopoietic stem cells for autoantigen recognition. This step can be determined according to the actual situation at the time of treatment.
  • the abort factor can be input as needed to enable early immune recognition of hematopoietic stem cells and/or late immune recognition hematopoietic cells to stop autoantigen immune recognition.
  • Figure 5 is a flow chart showing an experimental method for early immune recognition of hematopoietic stem cells and/or late immune recognition hematopoietic stem cells of the present invention in a mediator.
  • Performing an early immunological identification of hematopoietic stem cells and/or a latent immune recognition hematopoietic stem cell extraction, culture, amplification, and mediator experiment for secondary immune recognition in a mediator can use the following method: The mediator is divided into two parts and cultured simultaneously. .
  • the first group is the transplant donor's graft or graft antigen to the mediator's body group.
  • the intermediary used can use a. immunodeficient animals, such as: nude mice, SCID mice, NOD-SCID mice, etc., b. non-immune-deficient animals, for myeloablative, semi-myeloablative or non-myeloablative treatment, Early immune recognition of hematopoietic stem cells and/or advanced immune recognition of hematopoietic stem cell bone marrow transplanted animals can be performed.
  • Transplant donor or/or recipient graft or graft antigen into the body of the intermediary can recognize the donor and/or recipient's own antigen as self-antigens when performing self-antigen recognition. Since the recipient's non-immune recognition stem cells themselves also carry the recipient's own antigen and have strong expression, it is possible to experimentally determine whether or not to additionally transplant the recipient's graft antigen into the intermediary.
  • Transplant immune organs into the intermediary such as transplanted human thymus, spleen, human bone marrow tissue, organs and other organs, or cells in these tissues and organs related to autoimmune recognition. This step of transplanting the immune organ can be determined according to the test requirements.
  • initiation factor and/or termination factor tissue, organ, cell, factor associated with autoantigen immune recognition
  • transplant or input the "organisms, organs, cells, and factors related to the autoimmune recognition of autoantigens" of the same species.
  • species, organs, cells, and factors related to immune recognition of autoantigens of different species.
  • the intermediary becomes: If the graft is human, such as a transplanted human hematopoietic stem cell, the animal at this time has a human or an animal that has undergone an immune system that has been identified by the graft or graft antigen. This animal with human immune system for graft or graft antigen recognition can be used not only for this experiment, but also for experiments of various types and purposes.
  • the extracted non-immune-recognized human stem cells are transplanted into a second group of intermediaries that have undergone autoantigen immune recognition.
  • Group 2 The graft or graft antigen of the donor is not transplanted into the body of the intermediary.
  • the mediator used can use a. immunodeficient animals, such as: nude mice, SCID mice or NOD-SCID mice, etc.
  • the experiment can be carried out by taking SCID mice or NOD-SCID mice as an example.
  • Immune organs are transplanted into the intermediary, such as transplanted homologous animals, or xenogeneic animals, or human thymus, spleen, human bone marrow tissue, and the like.
  • the intermediary becomes: If the graft is human, then the animal has the human immune system at this time (the donor has not been graft-immunized). This animal with human immune system can be used not only for this experiment, but also for experiments of various types and purposes.
  • the transplantation of the graft is performed at this time for tissue-related matching, only HLA-matched grafts, or HLA half. Matching can be successfully transplanted.
  • HLA semi-matched grafts were used for transplantation, and anti-rejection drugs were administered after transplantation.
  • a human bone marrow transplant is performed without immunologically recognized human stem cells.
  • early immune recognition hematopoietic stem cells and/or late immune recognition hematopoietic stem cells can be obtained using the method mentioned in the description of Fig. 3.
  • the immune organs used for immune organ transplantation can be used either human or animal.
  • Figure 6 is a flow chart showing another method for extracting, culturing and amplifying hematopoietic stem cells and/or advanced immune recognition hematopoietic stem cells for early bone marrow transplantation in accordance with the present invention.
  • Stillbirth from abortion Stripping can extract the organs needed for secondary immune recognition: thymus, bone, liver. Strip the liver, thymus and bone of the stillbirth, because the thymus and bones are easier to obtain, so
  • the tissues, organs, cells, and factors involved in the immune recognition of autoantigens are preferably obtained from the thymus and bones. Because the fetus is liver hematopoietic, early immune recognition hematopoietic stem cells and/or late immune recognition hematopoietic stem cells can be obtained from the liver, or early immune recognition hematopoietic stem cells and/or late immune recognition hematopoietic stem cells can be obtained from other organs or tissues. .
  • the fetal liver is made into a single cell suspension, and a nucleated cell containing a relatively high concentration of early immunologically recognized hematopoietic stem cells and/or a late immune recognition hematopoietic stem cell is extracted by density gradient centrifugation.
  • the thymus and bone can be directly transplanted into the body, and can also be made into a thymus cell suspension, and a bone marrow cell suspension. It is also possible to extract a tissue related to autoantigen immune recognition by using a special cell separation method such as an immunomagnetic bead.
  • the desired cells related to the immune recognition of the autoantigen in the organs, cells, and factors are further cultured or used directly.
  • Density gradient centrifugation can be used to extract nucleated cells containing high concentrations of early immune recognition hematopoietic stem cells and/or late immune recognition hematopoietic stem cells, or to further utilize immunomagnetic beads-flow cytometry
  • the method selects early immune recognition hematopoietic stem cells and/or late immune recognition hematopoietic stem cells from fetal liver cell suspension, for example, monoclonal antibody using stem cell antigen (Stemcellantigen, Sea) specifically expressed on the surface of stem cells and coated with A second antibody on the surface of the magnetic particle is separated from the hematopoietic stem cell which is not immunologically recognized in the bone marrow by a magnetic adsorption cell sorting method (MACS:).
  • stem cell antigen Ste cell antigen, Sea
  • Thymocytes and bone marrow cells required for immune recognition were extracted from thymus and bone marrow suspension.
  • the sorted thymocytes, bone marrow cells, early immune recognition hematopoietic stem cells, and/or late immunorecognition hematopoietic stem cells are cultured in a culture medium containing a combination of different growth factors for amplification, and may be a serum-free culture solution.
  • the currently identified stimuli and inhibitors that act on the expansion (proliferation) of hematopoietic stem cells are quite numerous, but only a few cytokines can function alone.
  • hematopoietic stem cell growth factors can be added to serum-free medium, such as SCF, IL-3, IL-6, TP0, Flt3-L, G-CSF and Epo, and other hematopoietic growth factors are combined to expand in vitro.
  • SCF serum-free medium
  • IL-3 IL-6
  • TP0 Flt3-L
  • G-CSF G-CSF
  • Epo hematopoietic growth factors
  • BMSCs can be co-cultured with hematopoietic stem cells.
  • Hematopoietic stem cells have the effect of enhancing amplification.
  • the nucleated cell number (NC), surface antigen, and colony forming ability were monitored.
  • the in vitro expansion culture is generally carried out for about 2 weeks. Of course, you can increase or decrease the number of days according to the actual situation.
  • Figure 7 is a flow chart showing the treatment of adult autoantigen re-identification and immune system diseases in another organ transplant patient and immune system disease patient of the present invention.
  • This organ transplant surgery can be performed before the secondary antigen recognition of the autoantigen, or the graft antigen can be transplanted into the body first, and the secondary antigen recognition of the autoantigen is first performed, and the graft antigen to be transplanted is recognized as the autoantigen, and the organ transplantation is performed. Surgery, which eliminates the need for tissue-associated antigen matching and prevents postoperative rejection.
  • Non-immune-recognized stem cell bone marrow transplantation is prepared after transplantation of the host patient or in the case of a patient such as an immune system disease.
  • Non-specific clearance, and / or specific immune blood method can be used to remove immune-related cells, including: “autoantigen immune recognition library” and cells storing the autoantigen immune recognition library, as well as peripheral blood and immune tissue.
  • Immune cells including: T cells and B cells that have undergone negative selection and positive selection in the thymus and bone marrow.
  • the original hematopoietic stem cells in the body can be killed without using drugs and/or radiation, and the early immune recognition hematopoietic stem cells and/or late immune recognition hematopoietic stem cells can be directly And/or "organisms, organs, cells, factors related to immune recognition of autoantigens" are entered into the body.
  • the immune recognition of autoantigens is a factor such as "autoantigen immune recognition factor” in "organizations, organs, cells, and factors related to autoimmune recognition of autoantigens" and/or various substances required for autoimmune recognition.
  • autoantigen immune recognition factor The role of early immune recognition of hematopoietic stem cells and/or late immune recognition of hematopoietic stem cells. Perform self-antigen immune recognition. In the middle, it is necessary to observe the time required for the reconstruction of the immune system. At this time, it is necessary to track various immunological test indicators.
  • Figure 8 Is an extraction process of hematopoietic stem cells, thymocytes, and bone marrow cells of one embodiment of the present invention: Fetus: 3 to 4 months old
  • the thymus is ground into a single cell solution in a serum-free cell culture medium, and the thymus is gently ground by a tissue grinder to prepare a thymus cell suspension, and the bone marrow cells are washed out with the culture solution.
  • the above three cells (thymus, bone marrow, liver) were adjusted to a concentration of 5 ⁇ 10 6 with a culture solution containing 10% DMS0, and stored in liquid nitrogen.
  • Figure 9 is an illustration of an autoantigen re-identification and immune system disease of an organ transplant in an animal of the present invention.
  • tissue, organs, and cells related to immune recognition of autoantigens can be transplanted in the body of an animal having certain immune function.
  • "Factors” include: thymus, thymocytes, bone marrow, bone marrow cells, and immune recognition of hematopoietic stem cells to obtain experimental animals with human immune system functions.
  • This animal with human immune function because of its human immune system, can Substituting the human body for various medical experiments, so that many scientific experiments that cannot be performed because of certain damage to the human body can be tested on such animals, and have the same effect as human experiments, because the St. Jude Children's Research Hospital of the United States ( St. Jude Children's Research Hospital, published in the May 15, 2005 issue of The journal of Immunology P6540-6545, describing them in
  • NOD/LtSz-scid IL2R Y nu11 transplanted human hematopoietic stem cells into an experimental animal model of the human immune system.
  • NOD/LtSz-scid IL2R Y " u " mice are NK cells that are non-functional and cannot kill transplanted human hematopoietic stem cells, so this mouse does not have any immune function and is rejection of the graft. , is completely acceptable.
  • NOD/LtSz-scid IL2R y nu mice with no immune function were used as experimental group and NOD-scid mice with certain immune function as control group, and NOD/LtSz- scid IL2R y nu11 was small.
  • the human hematopoietic cell transplantation was successful, and the NOD/LtSz- scid IL2R y nu11 mice obtained human immune cells, which have human immune function, while the NOD-scid mice with certain immune function have hematopoiesis.
  • the failure of stem cell transplantation, the result of hematopoietic stem cell transplantation failure of the NOD-scid mouse can be used as an experimental control group of the present invention, and it is proved that the method used in the present invention can be used in the animal having normal immune function by using the method of the present invention. Animals with human immune function are made.
  • the present invention uses a NOD-scid mouse having NK cell function (having a certain immune function), and usually requires human leukocyte antigen (HLA) matching when performing hematopoietic stem cell transplantation in a human body, and a semi-matched transplantation is required. Take anti-rejection drugs, otherwise the transplant will not be successful.
  • HLA human leukocyte antigen
  • MHC major histocompatibility antigen complex
  • the "tissue, organ, cell, and factor related to autoantigen immune recognition" injected into mice include: Thymus, thymocytes, bone marrow, bone marrow cells, and immunologically recognized hematopoietic stem cells are a very strong antigenic graft, and no anti-rejection drugs are administered to mice during transplantation, and this transplantation method is equal to Graft antigen transplantation is performed simultaneously with hematopoietic cell transplantation.
  • the successful transplantation of this example proves that the present invention can indeed carry out the immune system between different kinds of animals of the same species, and between humans and animals. The organs move to each other and overcome the rejection reaction. The correctness of the theory and practice of the present invention is demonstrated.
  • the present invention can be used in a variety of animals having immune functions by transplanting human "organisms, organs, cells, and factors related to autoantigen immune recognition" including: thymus, thymus cells, bone marrow, bone marrow cells, and immunity.
  • human "organisms, organs, cells, and factors related to autoantigen immune recognition" including: thymus, thymus cells, bone marrow, bone marrow cells, and immunity.
  • the experimental basis of this method is to identify hematopoietic stem cells and obtain human immune cells, experimental animals with human immune function.
  • organs, organs, cells, and factors related to autoantigen immune recognition between animals of different kinds of animals, including: thymus, thymocytes, bone marrow, bone marrow cells, and immune recognition hematopoietic stem cell transplantation,
  • the recipient has an animal that functions as a donor immune system.
  • the practice of the present invention demonstrates that a human immune system can be established in an animal having immune function, a human immune system test can be performed in place of a human, and human tissues, cells, organs, and the like can be cultivated in the body of the animal.
  • the "organisms, organs, cells, and factors related to the autoimmune recognition of the autoantigens" of the same species should be transplanted, because the mice were only about 29 days from fertilization to birth, and because the mice were The period of self-antigen immune recognition is not clear in the first few days, and it is only necessary to transplant human "organisms, organs, cells, and factors related to autoantigen immune recognition", which has a great influence on the results of the experiment, but the test The results can also demonstrate the correctness of the principles of the present invention.
  • the present invention is an embodiment designed by using an early immune recognition hematopoietic stem cell and/or a late immune recognition hematopoietic stem cell obtained by various methods, and performing adult self antigen re-identification and treatment of an immune system disease, but it is also possible to utilize each Early stage of development, various types of stem cells obtained by transformation, early identification of hematopoietic stem cells and / or late immune recognition of hematopoietic stem cells, stem cells that have been immunologically recognized for various types of diseases.

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Abstract

L'invention concerne un procédé d'obtention et d'application des matières requises pour la reconnaissance immunologique d'autoantigènes après la naissance pour traiter le rejet et les maladies du système immunitaire. En particulier, l'invention concerne un procédé de fabrication et d'obtention de cellules souches hématopoïétiques de reconnaissance immunologique jeunes et/ou des cellules souches hématopoïétiques de reconnaissance immunologique avancées requises pour la reconnaissance immunologique d'autoantigènes; et d'obtention de tissus et organes liés au système immunitaire de la période de reconnaissance immunologique d'autoantigènes, comprenant : les cellules et facteurs, etc., liés à la reconnaissance immunologique d'autoantigènes dans le thymus et la moelle. Le procédé servant à traiter le rejet et des maladies du système immunitaire effectue la reconnaissance immunologique d'autoantigènes vis-à-vis de chaque antigène de tissu, d'organe ou de greffe avec des variations d'autoantigènes. Le présent procédé peut non seulement traiter de nombreuses maladies liées à des troubles du système immunitaire et le rejet après une greffe d'organes, mais également traiter toutes sortes de maladies pour différents organes des récepteurs. La présente invention peut être appliquée non seulement à des êtres humains, mais également à des animaux.
PCT/CN2005/001842 2004-11-04 2005-11-04 Procédé d'obtention et d'application des matières pour la reconnaissance immunologique d'autoantigènes WO2006047954A1 (fr)

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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1058991C (zh) * 1992-07-10 2000-11-29 匹兹堡大学 造血促进细胞
WO2003022296A1 (fr) * 2001-09-07 2003-03-20 The Trustees Of Boston University Procede et composition pour traiter les troubles lies aux complexes immuns
WO2004024902A1 (fr) * 2002-09-16 2004-03-25 The Walter And Eliza Hall Institute Of Medical Research Procede pour traiter une maladie auto-immune
WO2004034966A2 (fr) * 2002-08-06 2004-04-29 Loma Linda University Substances pour prevenir et traiter les maladies auto-immunes

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1058991C (zh) * 1992-07-10 2000-11-29 匹兹堡大学 造血促进细胞
WO2003022296A1 (fr) * 2001-09-07 2003-03-20 The Trustees Of Boston University Procede et composition pour traiter les troubles lies aux complexes immuns
WO2004034966A2 (fr) * 2002-08-06 2004-04-29 Loma Linda University Substances pour prevenir et traiter les maladies auto-immunes
WO2004024902A1 (fr) * 2002-09-16 2004-03-25 The Walter And Eliza Hall Institute Of Medical Research Procede pour traiter une maladie auto-immune

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
ZHOU ZHIHAI ET AL: "Systemic lupus and peripheral blood stem cell transplantation", JTMU, vol. 9, no. 3, 2003, pages 445 - 448 *

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