Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Conditional suicide anti-tumor targeted immune cell and preparation method thereof

An immune cell, suicidal technology, applied in the field of immune cells, can solve the problems of poor controllability, high cost, short survival time in the body, etc., to avoid potential side effects and solve the effects of large side effects

Inactive Publication Date: 2020-01-07
苏州博棠再生医学科技有限公司
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In view of the above-mentioned deficiencies in the prior art, the purpose of the present invention is to provide a conditional suicide anti-tumor targeted immune cell and its preparation method, aiming to solve the problem of large side effects of tumor cell-targeted therapy in the prior art, which cannot be widely used clinically, Problems of short survival time in vivo, high cost, and poor controllability

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Conditional suicide anti-tumor targeted immune cell and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Preparation of lentiviral particles

[0055] Firstly, use Clontech's lentiviral vector pLVX-IRES-ZsGreen1, which not only contains multiple cloning sites, but also carries the green fluorescent marker gene ZsGreen. The SV40LT gene is cloned into the vector gene at BamHI and EcoRI sites by DNA recombination. At the same time, the plasmid pLVX-SV40LT-ZsGreen that can be packaged into lentiviral particles was obtained, and the plasmid pLVX-SV40LT-ZsGreen was packaged to a titer of not less than 10 with a lentivirus packaging system. 7 / ml lentiviral particles.

Embodiment 2

[0057] Preparation of immortalized T cells

[0058] Collect 10 mL of human peripheral blood, and obtain monocytes (PMBC) in human peripheral blood by Ficoll method. The specific operation is as follows:

[0059] Transfer 10mL whole blood into a 50mL centrifuge tube, add 10mL PBS solution to dilute and mix gently; Take two 15mL centrifuge tubes, first add 5mL 9% Ficoll solution, and then gently add 10mL diluted blood to two tubes The upper layer of the Ficoll of the centrifuge tube, avoid mixing the two solutions, centrifuge at 2000rpm for 20min, white layered cells appear, then use a pipette to pipette the layer of cells into another clean 15mL centrifuge tube, and add PBS to 15mL, 1500rpm After centrifugation for 10 minutes, remove the supernatant, then add RPMI1640 medium containing 10% FBS, 1% P / S to 15 mL, centrifuge at 1500 rpm for 10 minutes, remove the supernatant, and then add 10 mL of RPMI1640 containing 10% FBS, 1% P / S to culture Mononuclear cells (PMBC) are obtained by ...

Embodiment 3

[0062] Preparation of immortalized NK cells

[0063] First, collect 10 mL of human peripheral blood and obtain the monocytes (PMBC) in the human peripheral blood by the Ficoll method. The specific operations are as follows:

[0064] Transfer 10mL whole blood into a 50mL centrifuge tube, add 10mL PBS solution to dilute and mix gently; Take two 15mL centrifuge tubes, first add 5mL 9% Ficoll solution, and then gently add 10mL diluted blood to two tubes The upper layer of the Ficoll of the centrifuge tube, avoid mixing the two solutions, centrifuge at 2000rpm for 20min, white layered cells appear, then use a pipette to pipette the layer of cells into another clean 15mL centrifuge tube, and add PBS to 15mL, 1500rpm After centrifugation for 10 minutes, remove the supernatant, then add RPMI1640 medium containing 10% FBS, 1% P / S to 15 mL, centrifuge at 1500 rpm for 10 minutes, remove the supernatant, and then add 10 mL of RPMI1640 containing 10% FBS, 1% P / S to culture Mononuclear cells (P...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
diameteraaaaaaaaaa
diameteraaaaaaaaaa
Login to View More

Abstract

The invention discloses a conditional suicide anti-tumor targeted immune cell and a preparation method thereof, which includes the steps of: A. Using DNA recombinant technology to insert the cell immortalization gene into the cloning site of the first cloning vector and prepare virus particles; B. . Use magnetic beads to specifically select an immune cell in human peripheral blood, and then use the virus particles prepared in step A to infect to obtain immortalized immune cells; C. Use DNA recombinant technology to insert the suicide gene into the second cloning vector. into the prepared immortalized immune cells to obtain conditional suicide immune cells; D. Use DNA recombinant technology to insert the single-chain antibody gene into the cloning site of the third cloning vector and make virus particles, Conditioned suicidal immune cells are then transfected to obtain conditional suicidal anti-tumor targeting immune cells. It solves the problems in the existing technology of tumor cell targeted therapy with large side effects, inability to be widely used clinically, short in vivo survival time, high cost, and poor controllability.

Description

Technical field [0001] The invention relates to the field of immune cells, in particular to a conditional suicide anti-tumor targeted immune cell and a preparation method thereof. Background technique [0002] Immune cells have immune surveillance and killing effects on tumor cells, especially immune cells carrying tumor-specific antigen-presenting single-chain antibodies (ScFV) can efficiently and specifically recognize, bind and kill related tumor cells. For example, single-chain antibody T lymphocyte technology (CAR-T) with tumor-specific antigen presentation in the treatment of acute lymphoblastic leukemia and lymphoma, the remission rate of tumor patients is as high as 80%. However, traditional CAR-T cell technology has potential side effects such as cytokine storm. If this side effect cannot be effectively controlled, the probability of death of tumor patients is 5-10%; on the other hand, conventional CAR-T cell technology requires individuality The chemical preparation of...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/10C12N15/867C12N15/63
CPCC12N5/0636C12N5/0646C12N15/63C12N15/86C12N2510/04C12N2501/51C12N2501/515C12N2501/2302C12N2740/15043A61K39/464499A61K39/4611A61K2239/38A61K39/4613A61K2239/31
Inventor 刘小青陈光风史秀娟朱良
Owner 苏州博棠再生医学科技有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com