Magnetotactic immune cells and construction method and application thereof

An immune cell and magnetic technology, applied in the field of magnetotactic immune cells and their construction, can solve the problems of inability to enhance the penetration and retention of immune cells, restrict the enrichment efficiency of immune cells, and time-consuming and labor-intensive operation procedures, and achieve low toxicity and magnetic The effect of high responsiveness and high sensitivity

Inactive Publication Date: 2016-09-21
XINXIANG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

To this end, on the one hand, with the help of gene transfection technology, the corresponding co-stimulatory signal components are introduced into T cells in vitro, so as to prolong the survival time and therapeutic effect of T cells in vivo after reinfusion, but the technical means are cumbersome and the operation process is time-consuming. It is time-consuming and costly; on the other hand, it uses targeting technology, including passive targeting and active targeting, but passive targeting is disabled due to the inability to enhance immune cell penetration and retention in some tumors lacking blood vessels, while The strict one-to-one correspondence between receptors and ligands also limits the generalizability of active targeting
The above-mentioned defects all restrict the enrichment efficiency of immune cells in the tumor site, reducing the effect of immunotherapy

Method used

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  • Magnetotactic immune cells and construction method and application thereof
  • Magnetotactic immune cells and construction method and application thereof
  • Magnetotactic immune cells and construction method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment one: Fe 3 o 4 Preparation of nanoparticles

[0039] Preparation of Fe by Polyol Method 3 o 4 Nanoparticles: Put 720mg of iron acetylacetonate in a 100mL three-neck bottle with a condenser, vacuumize and nitrogen gas repeatedly three times, then inject 40mL of triethylene glycol into the reaction system, stir magnetically, heat in a sand bath, and slowly raise the temperature to 180°C, keep warm for 30min; then rapidly raise the temperature to 278°C (the boiling point of triethylene glycol), and reflux for 30min to obtain a black magnetic fluid; cool to room temperature, and use a mixed solution of ethanol:ethyl acetate (volume ratio 1:10) Repeated washing three times, after magnetic separation, Fe with a particle size of 1-25nm 3 o 4 nanoparticles. Will Fe 3 o 4 Nanoparticles are re-dispersed in ethanol and stored at low temperature for future use.

Embodiment 2

[0040] Example 2: Pullulan modified Fe 3 o 4 Preparation of Magnetic Nanoparticles (MMNPs)

[0041] Take Fe 3 o 4 100mg of nanoparticles, dispersed in 10mL of n-hexane, disperse 300mg of pullulan with a molecular weight of 2 to 100,000 Daltons in 30 to 100mL containing 0.5 to 2.0wt% polyvinyl alcohol (PVA) and 0.2 to 2.0wt% In the aqueous solution of sodium dodecylbenzenesulfonate (SDBS), use a milk homogenizer to fully emulsify at a speed of 15,000 rpm for 2 minutes, then use a mechanical stirrer to stir at a speed of 600 rpm for 4 to 8 hours to volatilize n-hexane, The nanoparticles were solidified and formed, and then dialyzed with deionized water for 48 hours to obtain purified pullulan-modified Fe 3 o 4 magnetic nanoparticles.

[0042] By adjusting the Fe 3 o 4 The ratio of rice grains to polysaccharides, the concentration of emulsifiers and surfactants, the strength of emulsification, and the selection of dispersing solvents can regulate the particle size (tens o...

Embodiment 3

[0043] Example 3: Construction of magnetotactic T lymphocytes (mt-T)

[0044] 1. Isolation and expansion of human peripheral blood T lymphocytes

[0045] Human peripheral blood mononuclear cells were isolated by Ficoll density gradient centrifugation, separated and purified by nylon wool column and sorted by immunomagnetic beads, and CD3+ T cells were isolated, and then cultured in serum-free and low-sugar DMEM medium at 37°C, 5% CO 2 conditions for 72 hours. The isolated T cells have good activity, such as figure 1 with figure 2 shown.

[0046] 2. Construction of magnetotactic T lymphocytes (mt-T)

[0047] Take the density in the exponential growth period as 5~10×10 5 5 mL of T cell suspension per mL, at 37°C, 5% CO 2 conditions for 24 hours. The MMNPs prepared in Example 2 were dispersed in serum-free and low-sugar DMEM medium, and then added to the cultured T cell suspension. The final concentration of MMNPs in the T cell suspension was 20-100 μg / mL. At 37°C, 5% C...

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Abstract

The invention discloses magnetotactic immune cells. The magnetotactic immune cells comprise immune cells and magnetic nanoparticles taken into the immune cells. The magnetotactic immune cells disclosed by the invention can be effectively enriched in a tumor site to improve targeting of the cell immunotherapy.

Description

technical field [0001] The present invention relates to a targeted immune cell and its construction method and application, in particular to a magnetotactic immune cell and its construction method and application. Background technique [0002] Since the United States announced the launch of the Precision Medicine Initiative in early 2015, precision medicine has opened a new door of hope for the treatment of complex chronic diseases, and has attracted widespread attention from the medical and technological communities, as well as ordinary doctors and patients. In the treatment of malignant tumors, how to realize Precision Cell Immunotherapy (PCIT) has become a hot research topic at home and abroad. At present, the immunotherapy technology represented by chimeric antigen receptor-modified T cell (CAR-T) technology has made breakthroughs in the use of specific antigens of tumor cells to target and eliminate tumors, laying a solid foundation for the development of PCIT . It ca...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/078A61K35/17A61P35/00
CPCC12N5/0634A61K35/17C12N2500/24
Inventor 朱武凌陈红丽解丽芹南文滨张其清
Owner XINXIANG MEDICAL UNIV
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