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Mesenchymal stem cells (MSCs) secreting IL-6 antibody/CD20 antibody as well as construction method and application of MSCs

A technology of IL-6 and stem cells, applied in the field of biomedicine, to achieve the effect of reducing blood drug concentration, good therapeutic effect, and avoiding the problem of even distribution of blood drug concentration and peak-to-valley problems

Active Publication Date: 2021-06-18
GUANGZHOU FIRST PEOPLES HOSPITAL (GUANGZHOU DIGESTIVE DISEASE CENT GUANGZHOU FIRST PEOPLES HOSPITAL GUANGZHOU MEDICAL UNIV THE SECOND AFFILIATED HOSPITAL OF SOUTH CHINA UNIV OF TECH)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In view of the recurrence, refractory and side effects of the existing drugs for the treatment of diffuse large B-cell lymphoma, based on the research findings of the inventor's research team, it is believed that the treatment of refractory and relapsed DLBCL should take into account both tumor cell killing and Reversing two aspects of the microenvironment that promote tumor development and their synergistic effects

Method used

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  • Mesenchymal stem cells (MSCs) secreting IL-6 antibody/CD20 antibody as well as construction method and application of MSCs
  • Mesenchymal stem cells (MSCs) secreting IL-6 antibody/CD20 antibody as well as construction method and application of MSCs
  • Mesenchymal stem cells (MSCs) secreting IL-6 antibody/CD20 antibody as well as construction method and application of MSCs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Mesenchymal stem cells secreting IL-6 antibody and CD20 antibody and construction method thereof

[0035] 1. A method for constructing mesenchymal stem cells secreting IL-6 antibody and CD20 antibody, comprising the following steps:

[0036] 1. Construct the tandem double antibody target gene Tandab (IL-6Ab / CD20Ab) of IL-6 antibody and CD20 antibody light chain and heavy chain. The gene is composed of: CD33SP-VLCD20Ab-GGGGS-VHCD20Ab-3×GGGGS-VLIL-6Ab - GGGGS-VHIL-6Ab-6His.

[0037] 2. Tandab (IL-6Ab / CD20Ab) vector construction and virus packaging

[0038] Entrust Guangzhou Jingyuan Biotechnology Co., Ltd. to customize the carrier YOE-LV001-Tandab (IL-6Ab / CD20Ab), such as figure 1 As shown, it was co-transferred into 293T cells together with the virus packaging helper plasmid, the lentivirus was packaged, and the antibody titer was detected to construct stable MSCs, and Tandab(IL-6Ab / CD20Ab)-MSCs were obtained. The specific preparation method is as follows: ...

Embodiment 2

[0075] Example 2 Mesenchymal stem cells secreting IL-6 antibody and its construction method

[0076] Referring to Example 1, the method for preparing mesenchymal stem cells that secrete IL-6 antibody and CD20 antibody, the method for constructing mesenchymal stem cells that secrete IL-6 antibody in this example is different from Example 1 in that the customized carrier is different. The custom-made carrier in the preparation process of the mesenchymal stem cells secreting IL-6 antibody in this example is YOE-LV001-Tandab (IL-6Ab); finally, the mesenchymal stem cells capable of secreting IL-6 antibody, namely Tandab ( IL-6Ab)-MSCs.

Embodiment 3

[0077] Example 3 Mesenchymal stem cells secreting CD20 antibody and its construction method

[0078] Referring to the method for preparing mesenchymal stem cells secreting IL-6 antibody and CD20 antibody in Example 1, the method for constructing mesenchymal stem cells secreting CD20 antibody in this example is different from Example 1 in that the customized carrier is different. This implementation For example, the customized carrier in the preparation process of CD20 antibody-secreting mesenchymal stem cells is YOE-LV001-Tandab(CD20Ab); finally, the mesenchymal stem cells capable of secreting CD20 antibodies are Tandab(CD20Ab)-MSCs.

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Abstract

The invention belongs to the technical field of biological medicine, and particularly relates to mesenchymal stem cells (MSCs) secreting an IL-6 antibody and / or a CD20 antibody as well as a construction method and application of the MSCs. Human MSCs are transfected through a lentiviral vector carrying an IL-6 antibody gene and / or a CD20 antibody gene, the MSCs secreting the IL-6 antibody and / or the CD20 antibody are prepared, the MSCs are used for treating diffuse large B-cell lymphoma (DLBCL), the IL-6 antibody and the CD20 antibody are continuously and stably highly expressed in local DLBCL tumor tissue, the development of IL-6-mediated DLBCL can be effectively inhibited by utilizing the synergistic anti-tumor effect of the IL-6 antibody and the CD20 antibody, tumor cells are killed by utilizing cytotoxicity, the influence and side effects of direct injection of the antibody on normal tissues and cells of a human body are reduced, the problem of blood concentration fluctuation of in-vitro administration is solved, and a new scheme of a double-targeted therapeutic drug which takes double antibodies taking MSCs as the carrier and simultaneously acts on tumor cells and microenvironments is provided for relapse and refractory DLBCL.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and specifically relates to mesenchymal stem cells secreting IL-6 antibody / CD20 antibody, a construction method and application thereof. Background technique [0002] Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of non-Hodgkin's lymphoma, with more than 100,000 cases worldwide. The R-CHOP first-line chemotherapy regimen consisting of rituximab (CD20mAb), cyclophosphamide, doxorubicin, vincristine and prednisone improved the prognosis of DLBCL patients, but one-third of DLBCL patients still had Progression or relapse due to chemotherapy resistance. In clinical practice, increasing the dose of CD20mAb does not significantly improve the clinical efficacy of patients with refractory or relapsed DLBCL, and the dose is as large as 500mg / m 2 The CD20 mAb can only improve the prognosis of some elderly male patients, but it can cause severe neutropenia, thrombocytopenia, infectio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/867C12N5/10C07K16/24C07K16/28A61K39/395A61K47/46A61P35/00
CPCC12N15/86C12N5/0663C12N5/0667C12N5/0668C07K16/248C07K16/2887A61K47/46A61P35/00C12N2740/15043C12N2800/107C12N2510/02A61K2039/507
Inventor 魏亚明
Owner GUANGZHOU FIRST PEOPLES HOSPITAL (GUANGZHOU DIGESTIVE DISEASE CENT GUANGZHOU FIRST PEOPLES HOSPITAL GUANGZHOU MEDICAL UNIV THE SECOND AFFILIATED HOSPITAL OF SOUTH CHINA UNIV OF TECH)
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