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Immune effector cell conversion receptor

A technology of immune effector cells and receptors, which is applied in the fields of medical immunology and molecular biology, and can solve the problems of poor effector effect, unsatisfactory killing effect of target cells, short proliferation and duration, etc.

Pending Publication Date: 2020-07-07
SHANGHAI CELL THERAPY GRP CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The technical problem to be solved by the present invention is to overcome the technical problems of conventional immune effector cells such as T cells or TILs in the current field, which have short proliferation and duration in vivo, poor effect, and unsatisfactory killing effect on target cells. immune effector cell switching receptor

Method used

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Experimental program
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preparation example Construction

[0132]The method for preparing cells expressing immune effector cell switching receptors of the present invention includes the step of introducing the nucleic acid encoding immune effector receptors of the present invention into cells. This step can be performed ex vivo. For example, cells expressing the immune effector cell receptors of the present invention can be prepared in vitro by transducing viral vectors or non-viral vectors containing the nucleic acids of the present invention.

[0133] In the preparation method of the present invention, cells from mammals, such as cells derived from humans, or cells derived from non-human mammals, such as monkeys, mice, rats, pigs, horses, cows, etc., can be used. , sheep or dog cells.

[0134] In one embodiment, the mammal is a human.

[0135] The cells used in the preparation method of the present invention are not particularly limited, and any cells can be used. For example, cells collected, isolated or purified from bodily flu...

Embodiment 1

[0164] Example 1 Construction of expression vectors expressing EGFR switching receptors

[0165] 1. Entrust Shanghai Jierui Biological Company to synthesize the EGFP extracellular region gene, introduce a polyclonal restriction site (BglII-XbaI-EcoRI-BamHI) upstream of it, and insert a restriction site (SalI-NheI-HindIII- SpeI), which was loaded into the pNB328 vector cut with EcoR1+SalI (see CN201510638974.7 for the structure and sequence of pNB328, the entire content of which is incorporated herein by reference), to obtain the pNB328-EGFP vector

[0166] 2. Entrust Shanghai Jierui Biological Co., Ltd. to synthesize the EGFR switch receptor gene, which sequentially includes CD8 signal peptide, EGFR extracellular region, CD8 hinge region, CD8 transmembrane region, CD28 co-stimulatory region, and truncated IL-2Rβ chain intracellular region and the CD3ζ signal region containing the YRHQ motif, the schematic diagram of which is shown in figure 1 As shown, the sequence is shown i...

Embodiment 2

[0169] Example 2 Using Cetuximab and CD28 Monoclonal Antibody to Stimulate T Cells Expressing EGFR Switching Receptor

[0170] 1) Peripheral blood mononuclear cells (PBMCs) were isolated from the blood of donor subjects by Ficoll separation method. Cultivate PBMCs for 2-4 hours. The unattached suspension cells are initial T cells. Collect the suspension cells into a 15ml centrifuge tube, centrifuge at 1200rmp for 3min, discard the supernatant, add physiological saline, centrifuge at 1200rmp for 3min, discard the physiological brine, and repeat this step.

[0171] 2) Take two 1.5ml centrifuge tubes, numbered a and b, and add 5×10 6 Step 1) Centrifuge the initial T cells obtained at 1200rpm for 3min, discard the supernatant, add normal saline, centrifuge at 1200rpm for 3min, discard the normal saline, and repeat this step;

[0172] 3) Coat the six-well plate with a coating solution containing 5 μg / mL cetuximab (purchased from Merck) and 5 μg / mL anti-CD28 antibody (purchased fr...

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Abstract

The invention provides an immune effector cell conversion receptor, coding nucleic acid of the immune effector cell conversion receptor, a nucleic acid construct containing the coding nucleic acid, animmune effector cell capable of expressing the immune effector cell conversion receptor, a pharmaceutical composition containing one or more selected from the group consisting of the immune effectorcell conversion receptor, the coding nucleic acid, the nucleic acid construct and the immune effector cell, a preparation method for the immune effector cell capable of expressing the immune effectorcell conversion receptor, and an application of one or more selected from the group consisting of the immune effector cell conversion receptor, the coding nucleic acid, the nucleic acid construct, theimmune effector cell and the pharmaceutical composition in preparation of anti-cancer drugs. According to the invention, through the interaction between the immune effector cell conversion receptor and a binding protein thereof, cells capable of expressing the immune effector cell conversion receptor can be effectively stimulated; specifically, immune effector cells activate signaling of a downstream intracellular CD3 zeta region and an STAT5 transcription factor signaling pathway mediated by a white hormone receptor chain mediated STAT3 transcription factor and a CD3 zeta region STAT5 binding motif; proliferation and continuation of the cells can be effectively promoted; and a novel method is provided for stimulating and amplifying the cells in vivo and / or in vitro.

Description

technical field [0001] The invention relates to the fields of medical immunology and molecular biology, in particular to an immune effector cell conversion receptor. Background technique [0002] Immune cell therapy technology is an emerging treatment technology that has received extensive and deep attention in recent years, especially for use in cancer treatment. Adoptive cell transfer (ACT) therapy is an important method for the treatment of tumors by immune cell therapy. This method usually includes the input of immune effector cells whose surface receptors have been genetically engineered into the patient to be treated, and the immune effector cells whose surface receptors have been modified can specifically target and kill tumor cells, thereby achieving anti-tumor effects. Effect. The modified immune effector cells introduced into the human body can be autologous or allogeneic. Adoptive cell transfer has shown some efficacy in the treatment of solid tumors and is not...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/62C12N5/10A61K35/17A61P35/00
CPCC07K14/71A61K35/17A61P35/00C07K2319/02C07K2319/33C12N2510/00
Inventor 钱其军刘韬张洁黄晨金华君
Owner SHANGHAI CELL THERAPY GRP CO LTD
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