Highly effective method for amplifying activated lymphocyte and cultivation system

Abstract

The invention relates to a method for expanding activated lymphocyte, comprising the steps of allowing biologic sample containing lymphocytes to contact immobilized anti-CD 3 antibodies, then cultivating activated lymphocytes with culture mediums in different IL-2 concentrations to expand the activated lymphocytes. The invention also relates to a cultivation system used for expanding activated lymphocytes. The expanded activated lymphocytes can be applied in the immunotherapy of tumor, infectious diseases, and congenital or acquired immunodeficiency diseases.

Description

technical field

[0001] The invention relates to the field of immunotherapy, and specifically relates to a method and a culture system for expanding activated lymphocytes. Background technique

[0002] In recent years, due to the advancement of immunology, various immunotherapeutic methods have been tried for various immunodeficiencies caused by the reduction of the immune surveillance ability of the body. Various immunodeficiencies resulting from decreased immune surveillance capabilities include cancer (malignant tumor), congenital immunodeficiency, acquired immunodeficiency, and the like. One of the purposes of immunotherapy is to restore or / and enhance the ability of immune surveillance to treat these diseases.

[0003] As an immunotherapy for cancer, cell adoptive immunotherapy has become an important method to control tumor growth and even kill tumor cells. In the 1980s, it was found that some cells in peripheral blood mononuclear cells could be specifically activated...

Examples

Embodiment

[0117] The following examples serve to illustrate the invention and are not intended to limit the invention in any way.

[0118] 1. Cell activation:

[0119] First prepare a culture flask coated with the immobilized anti-CD3 antibody. Anti-CD3 antibody (Ortho) was diluted to 10 μg / ml in sterile phosphate sodium chloride buffer (PBS). Diluted anti-CD3 antibody was aseptically spread on the bottom of a culture flask (Costa). overnight at 4°C. Wash the unimmobilized anti-CD3 antibody with phosphate sodium chloride buffer solution (PBS), and then dry it naturally in a refrigerator at 4-10°C to obtain a carrier for T cell culture on which the anti-CD3 antibody immobilized surface is dried, and aseptically seal Afterwards, an activated culture vessel is obtained.

[0120] Collect 20ml of human venous whole blood with a syringe treated with heparin saline, add 20ml of normal saline, and carry out 2-fold dilution, and add 3ml of density gradient centrifuge Ficoll-Paque (Sigma-Aldric...

technical field

[0001] The invention relates to the field of immunotherapy, and specifically relates to a method and a culture system for expanding activated lymphocytes. Background technique

[0002] In recent years, due to the advancement of immunology, various immunotherapeutic methods have been tried for various immunodeficiencies caused by the reduction of the immune surveillance ability of the body. Various immunodeficiencies resulting from decreased immune surveillance capabilities include cancer (malignant tumor), congenital immunodeficiency, acquired immunodeficiency, and the like. One of the purposes of immunotherapy is to restore or / and enhance the ability of immune surveillance to treat these diseases.

[0003] As an immunotherapy for cancer, cell adoptive immunotherapy has become an important method to control tumor growth and even kill tumor cells. In the 1980s, it was found that some cells in peripheral blood mononuclear cells could be specifically activated...

Examples

Embodiment

[0117] The following examples serve to illustrate the invention and are not intended to limit the invention in any way.

[0118] 1. Cell activation:

[0119] First prepare a culture flask coated with the immobilized anti-CD3 antibody. Anti-CD3 antibody (Ortho) was diluted to 10 μg / ml in sterile phosphate sodium chloride buffer (PBS). Diluted anti-CD3 antibody was aseptically spread on the bottom of a culture flask (Costa). overnight at 4°C. Wash the unimmobilized anti-CD3 antibody with phosphate sodium chloride buffer solution (PBS), and then dry it naturally in a refrigerator at 4-10°C to obtain a carrier for T cell culture on which the anti-CD3 antibody immobilized surface is dried, and aseptically seal Afterwards, an activated culture vessel is obtained.

[0120] Collect 20ml of human venous whole blood with a syringe treated with heparin saline, add 20ml of normal saline, and carry out 2-fold dilution, and add 3ml of density gradient centrifuge Ficoll-Paque (Sigma-Aldric...

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