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Hla matching donor-originating activated lymphocytes

A lymphocyte and matching technology, applied in the direction of animal cells, vertebrate cells, cell culture active agents, etc., can solve unproven, patients cannot undergo bone marrow transplantation, side effects and other problems

Inactive Publication Date: 2004-03-17
LYMPHOTEC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, an HLA mismatch can lead to a fatal side effect, namely, fatal post-transfusion graft-versus-host disease (hereinafter referred to as "PT-GVHD") in patients after bone marrow transplantation or DLT therapy
[0008] Additionally, many patients cannot undergo bone marrow transplantation or blood transfusion due to manifested organ disease and infectious disease prior to organ transplantation or blood transfusion, even with an HLA-matched donor
Attempts to treat cancer patients who do not opt ​​for bone marrow transplantation by administering donor-derived lymphocytes have not proven to be very effective
[0009] In summary, the antitumor efficacy of autologous (patient-derived) or donor-derived lymphocytes is unsatisfactory when bone marrow transplantation is not an option, and their antiviral Effects and effects on autoimmune diseases are unclear
Another problem is that lymphocytes cannot be produced in large quantities without relying on a lymphocyte donor
Furthermore, DLI, which must be performed in conjunction with hematopoietic stem cell transplantation, is very limited as a treatment option because DLI cannot be performed if the patient cannot undergo hematopoietic stem cell transplantation, and because DLI must be performed in conjunction with stem cell transplantation , which requires a high HLA match rate, therefore, the probability of finding an HLA-matched donor is very low

Method used

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Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0061] (preparation kit for the preparation of preparations whose main components are activated lymphocytes derived from peripheral blood)

[0062] Although activated lymphocytes derived from HLA-matched donors can be used as a separate reagent in the present invention, they can also be prepared in the same flask with culture medium containing interleukin 2 and a flask containing solid-phase anti-CD3 antibody. in the kit. In the latter case, the formulations of the present invention can be prepared very easily using the kit.

[0063] It should be noted that an appropriate amount of culture solution for preparation of preparations can be poured into respective flasks coated with solid-phase anti-CD3 antibody in advance, and then these flasks can be stored in a refrigerator for frozen storage. The activated lymphocytes and at least two above-mentioned reagents are prepared in the same kit, and when activated lymphocytes are needed, the preparation of the present invention can b...

Embodiment 1

[0069] The first step, (collecting blood and separating lymphocytes)

[0070] Add heparin to the syringe, collect 20ml of peripheral blood from the vein of a donor whose HLA exactly matches the acute lymphoblastic leukemia patient, then pull out the needle from the syringe on a clean bench under sterile conditions and replace it with a 19G ×11 / 2″ injection needle. Inject 15ml of washing solution (RPMI 1640+6) (500ml, manufacturer: Nikken Biomedical) into two 50ml centrifuge tubes (manufacturer: Iwaki Glass Co., Ltd., 2341-050) in advance Research Center, GM1106). Then, dilute the collected blood with culture medium to triple the volume of the solution. Slowly pour the resulting mixture into the above two centrifuge tubes so that all the Pour in exactly equal amounts of the mixture.

[0071] Cover the lid of the centrifuge tube, turn over and shake two to three times, draw 3ml Lymphocepar 1 (100ml, manufacturer: Immunobiological Research Center Co., Ltd., 23010) with a 10ml st...

Embodiment 2

[0099] Activated lymphocytes were prepared as in Example 1, except that peripheral blood was obtained from blood relatives who showed microchimerism with patients with acute myeloid leukemia (2 HLA locus matches with patients). The preparation for supply was prepared as in Example 1, and the preparation was administered intravenously to patients with acute myeloid leukemia who had not undergone hematopoietic stem cell transplantation, and the preparation was only administered once. Three days after administration, the leukemic cells in the patient's peripheral blood were observed to decrease, and seven days later, the leukemic cells in the patient's peripheral blood completely disappeared. Therefore, it was confirmed that activated lymphocytes derived from donors with microchimerism had antitumor effects.

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PUM

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Abstract

According to the present invention, HLA matching donor-originating activated lymphocytes highly effective in the treatment of patients with tumors, viral infections and autoimmune diseases are prepared by collecting cells contained in peripheral blood originating from an HLA matching donor or peripheral blood originating from an HLA matching donor achieving micro-chimerism and by stimulating and propagating the cells thus collected with, for instance, interleukin 2 and anti-CD3 antibodies or the like. The HLA matching donor-originating activated lymphocytes thus prepared can be administered to an HLA matching patient with a tumor, a viral infection or an autoimmune disease for therapeutic or preventive purposes.

Description

technical field [0001] The present invention relates to activated lymphocytes derived from HLA-matched donors for the prevention and treatment of tumors, infectious diseases, and autoimmune diseases, a therapeutic method using said activated lymphocytes, the lymphocytes as a main component A formulation, a method for producing the formulation, and a kit for preparing the formulation. The object of the present invention is to more effectively treat and prevent tumors, such as cancer, and infectious diseases. Background technique [0002] Lymphocytes have received a lot of attention in recent years as a key biodefense measure in support of the immune system. T lymphocytes play an extremely important role in cell-mediated immunity. T lymphocytes are classified according to their reactivity with monoclonal antibodies, eg, T lymphocytes that show reactivity with anti-CD3 antibodies (CD means "cluster of differentiation") are classified as CD3 positive cells. So far, important ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/10A61K35/12A61K35/14A61K35/17A61K47/02A61K47/42A61P31/00A61P31/12A61P35/00A61P37/00A61P37/02C12N5/0783
CPCA61K2035/124C12N2501/515C12N2501/23C12N5/0636A61P31/00A61P31/12A61P35/00A61P37/00A61P37/02A61K35/14
Inventor 黒岩保幸森尾友宏清水则夫关根晖彬
Owner LYMPHOTEC
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