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Method for promoting CD4+T proliferation and activation and suppressing Jurkat T cell by using red cassia tree lectin

A technology of laurel wood lectin and CD4, which is applied in the field of lectins to achieve the effect of inhibiting proliferation and migration and maintaining biological activity

Inactive Publication Date: 2011-10-26
GUANGXI MEDICAL UNIVERSITY
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Problems solved by technology

[0006] After searching, there is no report that laurel wood agglutinin promotes the proliferation and activation of human peripheral blood CD4+ T lymphocytes, and inhibits the proliferation and migration of human acute lymphoblastic leukemia Jurkat T cells

Method used

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  • Method for promoting CD4+T proliferation and activation and suppressing Jurkat T cell by using red cassia tree lectin
  • Method for promoting CD4+T proliferation and activation and suppressing Jurkat T cell by using red cassia tree lectin
  • Method for promoting CD4+T proliferation and activation and suppressing Jurkat T cell by using red cassia tree lectin

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Embodiment 1

[0054] 1. Separation and purification of laurel lectin

[0055] All operations for separation and purification of laurel wood lectin were carried out at 4°C. Firstly take 30g of Osmanthus fragrans seeds, crush and grind them, add 150ml of 0.01mol / L phosphate buffer solution (pH7.2, containing 0.1mol / L NaCl) at a ratio of 1:5, stir with a magnetic stirrer for 2h, soak and extract for 24h. The coagulation activity of the extract is at 2 9 -2 11 between. The extract was filtered, the filtrate was centrifuged at 3000 rpm for 15 min, and ammonium sulfate was added to the supernatant to reach 30% saturation. Centrifuge at 3000rpm for 20min, and discard the precipitate. Ammonium sulfate was added to the supernatant to 60% saturation in the same way as above, and left for 12 hours. Centrifuge at 3000rpm for 20min. Discard the supernatant, and dissolve the precipitate with a small amount of 0.01mol / L phosphate buffer solution (pH7.2~7.4, containing 0.mol / L NaCl). Dialyze the cru...

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Abstract

The invention discloses a method for promoting cell proliferation and activation of peripheral blood CD4+T lymphocyte and suppressing cell proliferation and migration of acute lymphocytic leukemia (ALL) Jurkat T cells by using red cassia tree lectin. The separation and purification of red cassia tree lectin from red cassia tree seeds can be carried out at the temperature of 1-5 DEG C during the entire process, the proliferation and activation of the peripheral blood T lymphocyte CD4 subgroup can be promoted by the red cassia tree lectin through the verifications of an MTT (3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide) experiment, a DNA (deoxyribonucleic acid) analysis and an FCM (flow cytometry), and the red cassia tree lectin is CD4+T lymphocyte mitogen more efficient than PHA (Phytohaemagglutinin) and provides a new and effective way for anti-virus infection and vaccine intensifier development; through the verifications of CCK-8 (Cell Counting Kit-8) experiment, DNA analysis, cell apoptosis analysis, ELISA (enzyme-linked immuno sorbent assay) experiments and migration assay, the red cassia tree lectin can obviously suppress the proliferation and migration of the Jurkat T lymphocyte, is a growth inhibitor more efficient for the ALL cells and provides a new and effective supplementary means for treating the hematological malignancies.

Description

technical field [0001] The present invention relates to a lectin that inhibits the proliferation and migration of Jurkat T cells in human acute lymphoblastic leukemia, especially the laurel wood lectin that promotes the proliferation and activation of CD4+ T lymphocytes in human peripheral blood and inhibits the proliferation and migration of Jurkat T cells in human acute lymphoblastic leukemia Methods. Background technique [0002] Leukemia is a malignant disease of the hematopoietic system, also known as "blood cancer", and is one of the top ten malignant tumors with high incidence in China. It is characterized by the unlimited proliferation of a large number of leukemia cells in the bone marrow and other hematopoietic tissues, and enters the peripheral blood, while the production of normal hematopoietic cells and their normal biological functions are significantly inhibited. The disease ranks first in the malignant diseases of young people, especially children have becom...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783
Inventor 曾麒燕崔博王婧娉
Owner GUANGXI MEDICAL UNIVERSITY
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