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Adoptive-immunity cell for expressing recombinant T alpha 1 and preparation method and application of adoptive-immunity cell

An immune cell, secretory expression technology is applied in the field of adoptive immunotherapy cells expressing recombinant Tα1 and its preparation, which can solve the problems of small molecular weight of Tα1, short half-life in vivo, inconvenience for users, etc., and achieve the effect of improving immunity of the body

Inactive Publication Date: 2017-08-11
新钥(南京)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, so far, the products produced by genetic engineering technology have not been able to be used in clinical treatment.
[0005] In addition, Tα1 has a small molecular weight and a short half-life in the body. Clinical application requires repeated intravenous administration, which brings inconvenience to users and poor compliance.

Method used

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Examples

Experimental program
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Effect test

preparation example Construction

[0027] The present invention also provides the above-mentioned preparation method of adoptive immune cells expressing recombinant Tα1, comprising the following steps: culturing human immune cells in vitro, preparing gene vectors encoding recombinant hirudin and membrane-type molecular targets, the gene vector consists of signal peptide sequence- Tα1 sequence and signal peptide sequence-target molecule sequence-Linker-transmembrane sequence; the gene vector encoding recombinant thymosin α1 and target molecule is transfected into immune cells by genetic engineering technology.

[0028] Genetic engineering technology is a well-known technology, and the gene transfection system used includes lentiviral transfection system, retroviral transfection system, adenoviral transfection system, adeno-associated virus transfection system, Sleeping Beauty transposon transfection system, plasmid electroporation system, etc. Transfection system. Preferably, Sleeping Beauty transposon transfect...

Embodiment 1

[0033]The adoptive immune cells expressing recombinant Tα1, the adoptive immune cells are human immune cells, and the recombinant Tα1 expressed by the adoptive immune cells include human secreted expression signal peptide and human Tα1. The adoptive immune cells are one or more of αβT, γδT, NKT, NK, DC, CIK, CAR-T, CAR-NK, and TCR-T, and the adoptive immune cells are natural or artificially cultured immune cells. The preferred adoptive immune carrier cells are CIK, NK or γδT cells, which can simultaneously play the role of anti-aging, health care, anti-infection, and anti-tumor cellular immunotherapy.

[0034] The membrane surface of adoptive immune cells expresses target molecules, and the target molecules are located on the cell membrane surface through transmembrane sequences. The target molecule is the full length of CD19 or its truncated fragment, or the full length of CD20 or its truncated fragment.

[0035] Recombinant Tα1 is expressed constitutively or induced by a co...

Embodiment 2

[0040] The adoptive immune cells expressing recombinant Tα1 in this example are basically the same as in Example 1, except that the recombinant Tα1 expressed by adoptive immune cells is the fusion sequence PS-Tα1 of human amidase signal peptide and human Tα1, and its amino acid The sequence is: SEQ ID NO.3, and the gene sequence is: SEQ ID NO.4.

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Abstract

The invention discloses an adoptive-immunity cell for expressing recombinant thymosin alpha 1 and a preparation method and application of the adoptive-immunity cell. The adoptive-immunity cell is a human-derived immune cell, and the recombinant thymosin alpha 1 expressed by the adoptive-immunity cell comprises a signal peptide and thymosin alpha 1. The human-derived secreting signal peptide is connected to the amino terminal of the recombinant thymosin alpha 1, the encoding gene of the recombinant thymosin alpha 1 is used to transfect the adoptive-immunity cell through the genetic engineering technology, the recombinant thymosin alpha 1 can be continuously and stably expressed after the adoptive-immunity cell is transfused back into the body of a user, and a 'miniature cell pharmaceutical factory' effect is achieved. Accordingly, the recombinant thymosin alpha 1 can be stably expressed in the body of the user for a long time by using the adoptive-immunity cell as the carrier cell, and the defects that the thymosin alpha 1 is short in half-life period and high in preparation purity requirement, needs frequent injection causing inconvenience of patients, and the like are overcome.

Description

technical field [0001] The present invention relates to the fields of biotherapy and biomedicine, in particular to an adoptive immunotherapy cell expressing recombinant Tα1 and its preparation method and application. Background technique [0002] Thymosin is a general term for a class of polypeptide hormones produced by thymic epithelial cells that play an important role in the differentiation, maturation and immune activity of lymphocytes, also known as thymus hormone or thymosin. Thymosin α1 (Tα1) is a member of the thymosin α family. In recent years, it has attracted great attention due to its high biological activity and stability. Tα1 consists of 28 amino acids and is an immune enhancer against T cells. Tα1 molecule does not contain cysteine, so there is no disulfide bond. Tα1 has no glycosylation, and the only modification is acetylation at the N-terminal, but the presence or absence of acetyl group has no significant effect on the biological activity of Tα1. In vi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/85A61K35/17A61K35/15A61P1/16A61P35/00A61P31/00A61P39/06
Inventor 何向锋施文
Owner 新钥(南京)生物科技有限公司
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