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The method of improving the targeting force of NK cells and the application of ccr5 gene

A NK cell and gene technology, applied in the biological field, can solve the problems of weak directional movement ability of NK cells and restrict NK cell therapy, and achieve the effect of good clinical transformation prospects, strong tumor infiltration ability, and tumor growth inhibition effect.

Active Publication Date: 2022-06-24
THE FIRST AFFILIATED HOSPITAL OF ZHENGZHOU UNIV
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Problems solved by technology

However, the ability of traditionally applied NK cells to move toward tumor sites is weak, and only a small number of NK cells can infiltrate malignant tissues, which greatly limits NK cell therapy.

Method used

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  • The method of improving the targeting force of NK cells and the application of ccr5 gene
  • The method of improving the targeting force of NK cells and the application of ccr5 gene
  • The method of improving the targeting force of NK cells and the application of ccr5 gene

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Embodiment Construction

[0023] The experimental methods in the following examples are conventional methods unless otherwise specified, and the experimental reagents and materials involved are conventional biochemical reagents and materials unless otherwise specified.

[0024] 1. Acquisition of CCR5 coding sequence

[0025] The total RNA of human peripheral blood mononuclear cells was extracted with Trizol reagent (Invitrogen), and then reverse transcribed into cDNA. Then, PCR amplification was performed to obtain the coding sequence of CCR5 (as shown in SEQ ID NO. 1), wherein the upstream primer sequence: 5'- ggatcc atggattatcaagtgtcaag-3' (BamHI restriction site is underlined), downstream primer sequence: 5'- gcggccgc caagcccacagatatttc-3' (the NotI restriction site is underlined).

[0026] 2. Construction of CCR5 lentivirus overexpression plasmid

[0027] The PCR products of lentiviral plasmid pCDH-EF1-MSC-T2A-conGFP or CCR5 coding sequence were double digested with BamHI and NotI, respectively...

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Abstract

The invention belongs to the field of biotechnology, and specifically relates to a method for improving the targeting force of NK cells and the application of CCR5 gene in improving the specific motility of NK cells against tumors. The method can be highly expressed by transgenic CCR5 into NK cells NK of CCR5 will have stronger tumor-specific motility and better tumor control.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for improving the targeting force of NK cells and the application of CCR5 gene. Background technique [0002] Natural killer cells (NK) are a group of immune cells with strong tumor-killing ability and have good clinical application prospects. Clinical trials have demonstrated that NK cell infusion therapy can effectively control tumors and improve patient survival. However, the ability of traditionally applied NK cells to move toward tumor sites is weak, and only a small number of NK cells can infiltrate malignant tissues, which greatly limits NK cell therapy. Chemokines and their receptors play an important role in affecting the directional movement of immune cells to specific tissues. High expression of specific chemokine receptors can enhance the ability of NK cells to aggregate in specific tissues. Studies have shown that the chemokine CCL5 is highly expre...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/867C12N7/00C12N5/10C12R1/93
CPCC12N5/0646C12N7/00C12N15/86C12N2510/00C12N2740/15043C12N2740/15021
Inventor 盛誉乔李峰
Owner THE FIRST AFFILIATED HOSPITAL OF ZHENGZHOU UNIV
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