Chimeric antigen receptor gene for HIV-1, plasmid with gene, T cell, kit and application

A chimeric antigen receptor, HIV-1 technology, applied in the field of genes, can solve the problems that have not been widely used, the effect is not ideal, and CD4-ζCAR-T cells are susceptible to HIV-1 infection

Active Publication Date: 2018-05-01
SHANDONG XINRUI BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, although CD4-ζCAR-T cells can become activated T cells to kill HIV-1 infected cells after binding to gp120 antigen, it is difficult for activated T cells to proliferate further and can only

Method used

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  • Chimeric antigen receptor gene for HIV-1, plasmid with gene, T cell, kit and application
  • Chimeric antigen receptor gene for HIV-1, plasmid with gene, T cell, kit and application
  • Chimeric antigen receptor gene for HIV-1, plasmid with gene, T cell, kit and application

Examples

Experimental program
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Effect test

Embodiment 1

[0052] A chimeric antigen receptor gene for HIV-1, which includes a leader region, a domain that binds to the CD4 binding site of gp120, a hinge region, a transmembrane domain, a co-stimulatory domain, a signaling domain and Suicide gene system region. Wherein, the domain binding to the CD4 binding site of gp120 adopts any one of CD4 antigen, VRC01, VRC01b, VRC01-C, 3BNC60, and NIH45-46.

[0053] In this embodiment, as a preferred solution, the nucleotide sequence of the chimeric antigen receptor gene for HIV-1 is shown as SEQ ID NO.1. Namely: the following modules are concatenated to obtain: CD8 guider region nucleic acid artificial sequence, such as SEQ ID NO.2; CD4 antigen nucleic acid artificial sequence, such as SEQ ID NO.3; CD8 hinge region nucleic acid artificial sequence, such as SEQ ID NO.4 ; CD8 transmembrane region nucleic acid artificial sequence, such as SEQ ID NO.5; 4-1BB co-stimulatory region nucleic acid artificial sequence, such as SEQ ID NO.6; CD3ζ signaling...

Embodiment 2

[0055] Example of preparation of chimeric antigen receptor gene against HIV-1.

[0056] This embodiment is also illustrated by using the CD4 antigen as an example for the domain binding to the CD4 binding site of gp120. When using VRC01 (SEQ ID NO.10), VRC01b (SEQ ID NO.11), VRC01-C (SEQ ID NO.12), 3BNC60 (SEQ ID NO.13), and NIH45-46 (SEQ ID NO.14), the preparation method is basically the same, and will not be repeated here.

[0057] A method for preparing a chimeric antigen receptor gene for HIV-1, comprising the steps of:

[0058] (1) According to artificial nucleic acid sequence of CD8 leader region, artificial nucleic acid sequence of CD4 antigen, artificial nucleic acid sequence of CD8 hinge region, artificial nucleic acid sequence of CD8 transmembrane region, artificial sequence of nucleic acid of 4-1BB co-stimulatory region, artificial nucleic acid sequence of CD3ζ signal transduction region Artificial sequence, self-cleavage polypeptide T2A nucleic acid artificial seq...

Embodiment 3

[0065] A plasmid comprising a chimeric antigen receptor gene for HIV-1 as described above. The plasmid in this example is obtained by inserting the fusion gene fragment CD8leader-CD4-CD8-4-1BB-CD3ζ-T2A-iCasp9 into the lentiviral expression vector pLent-C-GFP.

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Abstract

The invention discloses an chimeric antigen receptor gene for HIV-1. The gene comprises a guide subregion, a structural domain, a hinge region, a trans-membrane structural domain, a co-stimulation structural domain, a signal transduction structural domain and a suicide gene system region, wherein the structural domain is combined with a CD4 binding site of a gp120. The invention further disclosesa plasmid with the gene, a T cell, a kit and application. According to the chimeric antigen receptor gene for the HIV-1, the plasmid with the gene, the T cell, the kit and the application, when the Tcell is applied, HIV-1 infection can be prevented, and meanwhile, the lethality on an HIV-1 infected cell can be improved; and once an HIV-1-CAR-T cell is input, adverse reactions occur, the cell in abody can be cleared at once through a suicide gene system, and therefore the preventive effect can be achieved.

Description

technical field [0001] The invention relates to the field of gene technology, in particular to a chimeric antigen receptor gene for HIV-1, a plasmid with the gene, T cells, a kit and application. Background technique [0002] Human immunodeficiency virus (human immunodeficiency virus Type 1, HIV-1), a type of retrovirus, invades CD4+ T cells, severely damages the body's acquired immune function, and eventually leads to AIDS (Acquired immunodeficiency syndrome). In August 2017, the Chinese Center for Disease Control and Prevention announced the national AIDS epidemic. The number of HIV-1 (human immunodeficiency virus) infections in China reached 718,270, of which 299,169 were suffering from AIDS, and the number of deaths due to AIDS has reached 221,628. . In addition, there are more than 140,000 new HIV-1 infected persons / AIDS patients each year. These figures show that the situation of AIDS infection in our country is very serious. [0003] At present, the clinical treatm...

Claims

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Application Information

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IPC IPC(8): C12N15/62C12N15/85C12N5/10A61K35/17A61P31/18
CPCA61K35/17C07K14/005C07K14/53C07K14/70514C12N5/0636C12N15/85C12N2510/00C12N2740/16122
Inventor 刘明录韩国英冯建海王立新金海锋万磊卢永灿韩庆梅刘敏马洪华强邦明张传鹏
Owner SHANDONG XINRUI BIOTECH CO LTD
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