CN111875710A - Immune cell for heterogeneous tumor treatment and application thereof - Google Patents
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Abstract
The invention provides an immune cell for heterogeneous tumor treatment and application thereof, wherein the immune cell expresses a chimeric antigen receptor which comprises an antigen binding domain, a transmembrane domain and a signal transduction domain; the antigen binding domain comprises NKG 2D; the signaling domains include 4-1BB, CD3 zeta, and TLR 2. The chimeric antigen receptor has a targeting effect on heterogeneous tumor cells, and the constructed immune cells expressing the chimeric antigen receptor can remove and kill the heterogeneous tumor cells by mobilizing the function of endogenous NKG 2D.
Description
Technical Field
The invention belongs to the technical field of biological medicines, and relates to immune cells for treating heterogeneous tumors and application thereof.
Background
A Chimeric Antigen Receptor (CAR) consists of a tumor-associated antigen binding region, an extracellular hinge region, a transmembrane region, and an intracellular signaling region. Generally, CAR molecules comprise an antibody single chain fragment variable (scFv) with specific binding for a Tumor Associated Antigen (TAA), which is coupled to the cytoplasmic domain of a signaling molecule via a hinge and a transmembrane region.
In recent years, with the development of tumor immunology theory and clinical technology, Chimeric antigen receptor T-cell immunotherapy (CAR-T) has become one of the most promising tumor immunotherapy. At present, CAR-T cell therapy has been widely applied to the treatment of B cell malignant tumors, and CAR-T cells targeting CD19 are precursors for the treatment of B cell malignant tumors by CAR-T therapy, and provide an effective scheme for the treatment of B cell malignant tumors. However, many tumor cells do not express specific, specific tumor antigens, and tumor heterogeneity is a major obstacle in the treatment of solid tumors by cell therapy.
Therefore, there is a need to develop chimeric antigen receptors for heterogeneous tumor cells to improve the recognition of non-specific antigen expressing tumors by the immune cells of the chimeric antigen receptors.
Disclosure of Invention
Aiming at the defects and actual requirements of the prior art, the invention provides the immune cell for treating the heterogeneous tumor and the application thereof, the immune cell realizes the effect of removing and killing the heterogeneous tumor cell by mobilizing the function of endogenous NKG2D, and has wide prospect in the aspect of treating the solid tumor.
In order to achieve the purpose, the invention adopts the following technical scheme:
in a first aspect, the present invention provides a chimeric antigen receptor comprising an antigen binding domain, a transmembrane domain, and a signaling domain;
the antigen binding domain comprises NKG 2D;
the signaling domains include 4-1BB, CD3 zeta, and TLR 2.
In the invention, NKG2D is used as an antigen binding domain, and is connected with a signal conduction domain to construct a chimeric antigen receptor, which has obvious targeting effect on tumor cells.
Preferably, the antigen binding domain comprises the amino acid sequence shown in SEQ ID NO. 1;
SEQ ID NO:1：
FNQEVQIPLTESYCGPCPKNWICYKNNCYQFFDESKNWYESQASCMSQNASLLKVYSKEDQDLLKLVKSYHWMGLVHIPTNGSWQWEDGSILSPNLLTIIEMQKGDCALYASSFKGYIENCSTPNTYICMQRTV。
preferably, the transmembrane domain comprises CD28 and/or CD8 a.
Preferably, the chimeric antigen receptor further comprises a signal peptide.
Preferably, the signal peptide comprises a GM-CSF signal peptide.
Preferably, the chimeric antigen receptor consists of a GM-CSF signal peptide, NKG2D, CD8 α, 4-1BB, CD3 ζ and TLR2 in tandem.
Preferably, the amino acid sequence of the chimeric antigen receptor is shown as SEQ ID NO. 2;
SEQ ID NO:2：
MLLLVTSLLLCELPHPAFLLFNQEVQIPLTESYCGPCPKNWICYKNNCYQFFDESKNWYESQASCMSQNASLLKVYSKEDQDLLKLVKSYHWMGLVHIPTNGSWQWEDGSILSPNLLTIIEMQKGDCALYASSFKGYIENCSTPNTYICMQRTVTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRQAKRKPRKAPSRNICYDAFVSYSERDAYWVENLMVQELENFNPPFKLCLHKRDFIPGKWIIDNIIDSIEKSHKTVFVLSENFVKSEWCKYELDFSHFRLFDENNDAAILILLEPIEKKAIPQRFCKLRKIMNTKTYLEWPMDEAQREGFWVNLRAAIKS。
in a second aspect, the present invention provides a coding gene encoding the chimeric antigen receptor of the first aspect.
Preferably, the coding gene comprises the NKG2D coding sequence.
Preferably, the coding gene further comprises a GM-CSF signal peptide coding sequence, a CD8 a coding sequence, a 4-1BB coding sequence, a CD3 ζ coding sequence and a TLR2 coding sequence.
Preferably, the coding gene comprises a nucleic acid sequence shown as SEQ ID NO. 3;
SEQ ID NO:3：
atgcttctcctggtgacaagccttctgctctgtgagttaccacacccagcattcctcctgttcaaccaagaagttcaaattcccttgaccgaaagttactgtggcccatgtcctaaaaactggatatgttacaaaaataactgctaccaattttttgatgagagtaaaaactggtatgagagccaggcttcttgtatgtctcaaaatgccagccttctgaaagtatacagcaaagaggaccaggatttacttaaactggtgaagtcatatcattggatgggactagtacacattccaacaaatggatcttggcagtgggaagatggctccattctctcacccaacctactaacaataattgaaatgcagaagggagactgtgcactctatgcctcgagctttaaaggctatatagaaaactgttcaactccaaatacgtacatctgtatgcaaaggactgtgaccacgacgccagcgccgcgaccaccaacaccggcgcccaccatcgcgtcgcagcccctgtccctgcgcccagaggcgtgccggccagcggcggggggcgcagtgcacacgagggggctggacttcgcctgtgatatctacatctgggcgcccttggccgggacttgtggggtccttctcctgtcactggttatcaccctttactgcaaacggggcagaaagaaactcctgtatatattcaaacaaccatttatgagaccagtacaaactactcaagaggaagatggctgtagctgccgatttccagaagaagaagaaggaggatgtgaactgagagtgaagttcagcaggagcgcagacgcccccgcgtaccagcagggccagaaccagctctataacgagctcaatctaggacgaagagaggagtacgatgttttggacaagagacgtggccgggaccctgagatggggggaaagccgagaaggaagaaccctcaggaaggcctgtacaatgaactgcagaaagataagatggcggaggcctacagtgagattgggatgaaaggcgagcgccggaggggcaaggggcacgatggcctttaccagggtctcagtacagccaccaaggacacctacgacgcccttcacatgcaggccctgccccctcgccaggccaaaaggaagcccaggaaagctcccagcaggaacatctgctatgatgcatttgtttcttacagtgagcgggatgcctactgggtggagaaccttatggtccaggagctggagaacttcaatccccccttcaagttgtgtcttcataagcgggacttcattcctggcaagtggatcattgacaatatcattgactccattgaaaagagccacaaaactgtctttgtgctttctgaaaactttgtgaagagtgagtggtgcaagtatgaactggacttctcccatttccgtctttttgatgagaacaatgatgctgccattctcattcttctggagcccattgagaaaaaagccattccccagcgcttctgcaagctgcggaagataatgaacaccaagacctacctggagtggcccatggacgaggctcagcgggaaggattttgggtaaatctgagagctgcgataaagtcc。
in a third aspect, the present invention provides an expression vector, wherein the expression vector is a lentiviral vector comprising the coding gene of the second aspect.
In a fourth aspect, the present invention provides a recombinant lentivirus which is a mammalian cell transfected with an expression vector and a helper plasmid as described in the third aspect.
In a fifth aspect, the present invention provides a chimeric antigen receptor immune cell expressing the chimeric antigen receptor of the first aspect.
Preferably, the chimeric antigen receptor immune cell has the coding gene of the second aspect integrated into its genome.
Preferably, the chimeric antigen receptor immune cell comprises the expression vector of the third aspect and/or the recombinant lentivirus of the fourth aspect.
Preferably, the chimeric antigen receptor immune cell is a T cell expressing the chimeric antigen receptor of the first aspect.
Preferably, the chimeric antigen receptor immune cell is an NK cell expressing the chimeric antigen receptor of the first aspect.
In a sixth aspect, the present invention provides a method for producing the chimeric antigen receptor immune cell of the fifth aspect, the method comprising the step of introducing the gene encoding the chimeric antigen receptor of the first aspect into an immune cell.
In a seventh aspect, the present invention provides a use of the chimeric antigen receptor of the first aspect, the coding gene of the second aspect, the expression vector of the third aspect, the recombinant lentivirus of the fourth aspect, or the chimeric antigen receptor immune cell of the fifth aspect in the preparation of a medicament for treating a disease.
Preferably, the disease comprises a tumor.
Preferably, the tumor includes any one or a combination of at least two of liver cancer, lung cancer, breast cancer, stomach cancer, nephroblastoma, glioma, neuroblastoma, melanoma, nasopharyngeal carcinoma, mesothelioma, islet cell tumor, retinoblastoma, pancreatic cancer, uterine fibroids, cervical cancer, or thyroid cancer.
Compared with the prior art, the invention has the following beneficial effects:
(1) the chimeric antigen receptor constructed by the invention adopts NKG2D as an antigen binding domain, is connected with a signal conduction domain to construct the chimeric antigen receptor, and has obvious targeting effect on tumor cells;
(2) the immune cell expressing the chimeric antigen receptor constructed by the invention has obvious effects of eliminating and killing heterogeneous tumor cells by exerting the effect of recognizing cell stress proteins such as MICA/B antigen, ULBP family antigen and the like by endogenous NKG2D receptor.
Drawings
Figure 1 is a schematic structural diagram of the NKG2D CAR molecule;
FIG. 2 shows the killing efficiency of WT, anti CD19-CAR-T and NKG2D-CAR-T on tumor cell BGC823 at different E: T ratios.
Detailed Description
To further illustrate the technical means adopted by the present invention and the effects thereof, the present invention is further described below with reference to the embodiments and the accompanying drawings. It is to be understood that the specific embodiments described herein are merely illustrative of the invention and are not limiting of the invention.
The examples do not show the specific techniques or conditions, according to the technical or conditions described in the literature in the field, or according to the product specifications. The reagents or apparatus used are conventional products commercially available from normal sources, not indicated by the manufacturer.
Example 1 construction of CAR molecular vectors
The NKG2D CAR molecule is constructed in the embodiment, the schematic structural diagram is shown in FIG. 1, the amino acid sequence is shown in SEQ ID NO. 2, and the nucleic acid sequence is shown in SEQ ID NO. 3;
firstly, synthesizing an encoding gene (SEQ ID NO:3) of the NKG2D chimeric antigen receptor by gene, and respectively adding a restriction enzyme Pme1 enzyme cutting site and a protective base thereof, and a restriction enzyme Spe1 enzyme cutting site and a protective base thereof at the 5 'end and the 3' end of the encoding gene;
the coding gene is subjected to double enzyme digestion by restriction enzymes Pme1 and Spe1, an enzyme digestion product containing a sticky end is obtained by agar gel electrophoresis recovery, and is connected into a linearized pWPXld-eGFP plasmid (containing the sticky end) which is also subjected to double enzyme digestion by Pme1 and Spe1, and the connection reaction is carried out in the presence of T4 DNA polymerase (Invitrogen company) to obtain a lentiviral vector containing the coding gene of the CAR molecule.
In this example, a CAR with an antigen-binding domain of anti-CD 19scFv (anti CD19scFv-CD8 α -4-1BB-CD3 ζ) was constructed at the same time, and a corresponding lentiviral vector was constructed.
Example 2 Lentiviral packaging
Transferring the lentiviral vector constructed in the embodiment 1 into escherichia coli, selecting positive monoclonal for overnight culture, extracting the lentiviral vector by using a plasmid extraction kit, and packaging viruses;
performing virus packaging by using 293T cells, preparing a packaging system, wherein a pWPXld-expression plasmid comprises a lentiviral vector containing an encoding gene of NKG2DCAR and a lentiviral vector containing an encoding gene of anti CD19 CAR, and the pWPXld-eGFP plasmid is an empty vector containing no CAR encoding gene;
the helper plasmid and the lentiviral vector with the CAR molecule are transfected into 293T cells at the same time, the first and second viruses are harvested after 48h and 72h respectively, and frozen at-80 ℃ after filtration for immune cell transduction.
Example 3T cell activation and lentivirus transfection
Isolating mononuclear cells (PBMCs) from adult peripheral blood, sorting out total T cells using a T cell sorting kit, and adding the CAR molecule over-expressing recombinant lentivirus prepared in example 2 after 24 hours of in vitro stimulation with CD3 and CD28 antibodies;
after 12h of transduction, the T cells were centrifuged and the solution was changed; three days after transduction, CAR-T ratios were assessed using flow cytometry to determine GFP positive cell content;
2X 10 cultures per ml of culture Medium6The CAR-T cells are expanded through density culture of the individual cells, NKG2D-CAR-T, antiCD19-CAR-T and WT (transfection blank control eGFP lentivirus) are obtained, the cells are frozen and stored for later use after ten days.
Example 4 functional assessment of NKG2D-CAR-T cells
This example used NKG2D-CAR-T cells prepared in example 3 to perform in vitro killing, and set negative control anti cd19-CAR-T cells and blank control WT, as follows:
firstly, counting gastric cancer target cells BGC823, NKG2D-CAR-T cells, anti CD19-CAR-T cells and WT cells, inoculating the CAR-T cells and the gastric cancer target cells in a white flat-bottom 96-well plate according to the proportion of 4:1, 2:1, 1:2, 1:4 and 1:8, and co-culturing for 24h by using 200 mu L of IMDM culture medium containing 5% fetal bovine serum;
after the co-culture is finished, 100 mu L of PBS containing Luciferase fluorogenic substrate is added into each hole, the luminous intensity is measured by using a multifunctional microplate reader, and a line graph of the in-vitro killing effect is obtained after calculation, and the in-vitro killing capacity of the NKG2D-CAR-T cells, the anti CD19-CAR-T cells and the WT cells to the gastric cancer target cells BGC823-GL is visually compared.
As shown in figure 2, NKG2D-CAR-T cells have obvious killing effect on target cells with positive NKG2D ligand expression, and the killing capability is obviously improved compared with negative control anti CD19-CAR-T and blank control WT.
In conclusion, the NKG2D is used as an antigen binding domain and is connected with a signal conduction domain to construct a chimeric antigen receptor, so that the chimeric antigen receptor has an obvious targeting effect on tumor cells; the immune cells expressing the chimeric antigen receptor have obvious effects of eliminating and killing heterogeneous tumor cells by exerting the effect of recognizing cell stress proteins such as MICA/B antigens, ULBP family antigens and the like through the endogenous NKG2D receptor.
The applicant states that the present invention is illustrated in detail by the above examples, but the present invention is not limited to the above detailed methods, i.e. it is not meant that the present invention must rely on the above detailed methods for its implementation. It should be understood by those skilled in the art that any modification of the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.
SEQUENCE LISTING
<110> Guangdong Shoutai biomedical science and technology Co., Ltd
<120> immune cell for heterogeneous tumor treatment and application thereof
<130>2020
<160>3
<170>PatentIn version 3.3
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Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu Asp Gln Asp Leu Leu
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Tyr Cys Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn Cys
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Cys Met Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu Asp
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His Ile Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile Leu
100 105 110
Ser Pro Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys Ala
115 120 125
Leu Tyr Ala Ser Ser Phe Lys Gly Tyr Ile Glu Asn Cys Ser Thr Pro
130 135 140
Asn Thr Tyr Ile Cys Met Gln Arg Thr Val Thr Thr Thr Pro Ala Pro
145 150 155 160
Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu
165 170 175
Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg
180 185 190
Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly
195 200 205
Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Lys
210 215 220
Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg
225 230 235 240
Pro Val Gln Thr Thr Gln Glu GluAsp Gly Cys Ser Cys Arg Phe Pro
245 250 255
Glu Glu Glu Glu Gly Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser
260 265 270
Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu
275 280 285
Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg
290 295 300
Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln
305 310 315 320
Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr
325 330 335
Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp
340 345 350
Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala
355 360 365
Leu His Met Gln Ala Leu Pro Pro Arg Gln Ala Lys Arg Lys Pro Arg
370 375 380
Lys Ala Pro Ser Arg Asn Ile Cys Tyr Asp Ala Phe Val Ser Tyr Ser
385 390 395 400
Glu Arg Asp Ala Tyr Trp Val Glu Asn LeuMet Val Gln Glu Leu Glu
405 410 415
Asn Phe Asn Pro Pro Phe Lys Leu Cys Leu His Lys Arg Asp Phe Ile
420 425 430
Pro Gly Lys Trp Ile Ile Asp Asn Ile Ile Asp Ser Ile Glu Lys Ser
435 440 445
His Lys Thr Val Phe Val Leu Ser Glu Asn Phe Val Lys Ser Glu Trp
450 455 460
Cys Lys Tyr Glu Leu Asp Phe Ser His Phe Arg Leu Phe Asp Glu Asn
465 470 475 480
Asn Asp Ala Ala Ile Leu Ile Leu Leu Glu Pro Ile Glu Lys Lys Ala
485 490 495
Ile Pro Gln Arg Phe Cys Lys Leu Arg Lys Ile Met Asn Thr Lys Thr
500 505 510
Tyr Leu Glu Trp Pro Met Asp Glu Ala Gln Arg Glu Gly Phe Trp Val
515 520 525
Asn Leu Arg Ala Ala Ile Lys Ser
530 535
<210>3
<211>1608
<212>DNA
<213> Artificial sequence
<400>3
atgcttctcc tggtgacaag ccttctgctc tgtgagttac cacacccagc attcctcctg 60
ttcaaccaag aagttcaaat tcccttgacc gaaagttact gtggcccatg tcctaaaaac 120
tggatatgtt acaaaaataa ctgctaccaa ttttttgatg agagtaaaaa ctggtatgag 180
agccaggctt cttgtatgtc tcaaaatgcc agccttctga aagtatacag caaagaggac 240
caggatttac ttaaactggt gaagtcatat cattggatgg gactagtaca cattccaaca 300
aatggatctt ggcagtggga agatggctcc attctctcac ccaacctact aacaataatt 360
gaaatgcaga agggagactg tgcactctat gcctcgagct ttaaaggcta tatagaaaac 420
tgttcaactc caaatacgta catctgtatg caaaggactg tgaccacgac gccagcgccg 480
cgaccaccaa caccggcgcc caccatcgcg tcgcagcccc tgtccctgcg cccagaggcg 540
tgccggccag cggcgggggg cgcagtgcac acgagggggc tggacttcgc ctgtgatatc 600
tacatctggg cgcccttggc cgggacttgt ggggtccttc tcctgtcact ggttatcacc 660
ctttactgca aacggggcag aaagaaactc ctgtatatat tcaaacaacc atttatgaga 720
ccagtacaaa ctactcaaga ggaagatggc tgtagctgcc gatttccaga agaagaagaa 780
ggaggatgtg aactgagagt gaagttcagc aggagcgcag acgcccccgc gtaccagcag 840
ggccagaacc agctctataa cgagctcaat ctaggacgaa gagaggagta cgatgttttg 900
gacaagagac gtggccggga ccctgagatg gggggaaagc cgagaaggaa gaaccctcag 960
gaaggcctgt acaatgaact gcagaaagat aagatggcgg aggcctacag tgagattggg 1020
atgaaaggcg agcgccggag gggcaagggg cacgatggcc tttaccaggg tctcagtaca 1080
gccaccaagg acacctacga cgcccttcac atgcaggccc tgccccctcg ccaggccaaa 1140
aggaagccca ggaaagctcc cagcaggaac atctgctatg atgcatttgt ttcttacagt 1200
gagcgggatg cctactgggt ggagaacctt atggtccagg agctggagaa cttcaatccc 1260
cccttcaagt tgtgtcttca taagcgggac ttcattcctg gcaagtggat cattgacaat 1320
atcattgact ccattgaaaa gagccacaaa actgtctttg tgctttctga aaactttgtg 1380
aagagtgagt ggtgcaagta tgaactggac ttctcccatt tccgtctttt tgatgagaac 1440
aatgatgctg ccattctcat tcttctggag cccattgaga aaaaagccat tccccagcgc 1500
ttctgcaagc tgcggaagat aatgaacacc aagacctacc tggagtggcc catggacgag 1560
gctcagcggg aaggattttg ggtaaatctg agagctgcga taaagtcc 1608
Claims (10)
1. A chimeric antigen receptor, comprising an antigen binding domain, a transmembrane domain, and a signaling domain;
the antigen binding domain comprises NKG 2D;
the signaling domains include 4-1BB, CD3 zeta, and TLR 2.
2. The chimeric antigen receptor according to claim 1, wherein the antigen binding domain comprises the amino acid sequence shown in SEQ ID No. 1.
3. The chimeric antigen receptor according to claim 1 or 2, wherein the transmembrane domain comprises CD28 and/or CD8 a;
preferably, the chimeric antigen receptor further comprises a signal peptide;
preferably, the signal peptide comprises a GM-CSF signal peptide.
4. The chimeric antigen receptor according to any one of claims 1 to 3, wherein the chimeric antigen receptor consists of a GM-CSF signal peptide, NKG2D, CD8 a, 4-1BB, CD3 ζ and TLR2 in tandem;
preferably, the amino acid sequence of the chimeric antigen receptor is shown as SEQ ID NO. 2.
5. A coding gene encoding the chimeric antigen receptor of any one of claims 1 to 4;
preferably, the coding gene comprises the NKG2D coding sequence;
preferably, the coding gene further comprises a GM-CSF signal peptide coding sequence, a CD8 a coding sequence, a 4-1BB coding sequence, a CD3 ζ coding sequence, and a TLR2 coding sequence;
preferably, the coding gene comprises a nucleic acid sequence shown as SEQ ID NO. 3.
6. An expression vector, wherein the expression vector is a lentiviral vector comprising the coding gene of claim 5.
7. A recombinant lentivirus, wherein the recombinant lentivirus is a mammalian cell transfected with the expression vector of claim 6 and a helper plasmid.
8. A chimeric antigen receptor immune cell, wherein said chimeric antigen receptor immune cell expresses the chimeric antigen receptor of any one of claims 1-4;
preferably, the chimeric antigen receptor immune cell has the coding gene of claim 5 integrated into its genome;
preferably, the chimeric antigen receptor immune cell comprises the expression vector of claim 6 and/or the recombinant lentivirus of claim 7;
preferably, the chimeric antigen receptor immune cell is a T cell expressing the chimeric antigen receptor of any one of claims 1-4;
preferably, the chimeric antigen receptor immune cell is an NK cell expressing the chimeric antigen receptor of any one of claims 1-4.
9. A method for producing the chimeric antigen receptor immune cell according to claim 8, which comprises the step of introducing the gene encoding the chimeric antigen receptor according to any one of claims 1 to 4 into an immune cell.
10. Use of the chimeric antigen receptor of any one of claims 1 to 4, the coding gene of claim 5, the expression vector of claim 6, the recombinant lentivirus of claim 7 or the chimeric antigen receptor immune cell of claim 8 for the preparation of a medicament for the treatment of a disease;
preferably, the disease comprises a tumor;
preferably, the tumor includes any one or a combination of at least two of liver cancer, lung cancer, breast cancer, stomach cancer, nephroblastoma, glioma, neuroblastoma, melanoma, nasopharyngeal carcinoma, mesothelioma, islet cell tumor, retinoblastoma, pancreatic cancer, uterine fibroids, cervical cancer, or thyroid cancer.
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Cited By (1)
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|---|---|---|---|---|
| CN114989310A (en) * | 2022-06-20 | 2022-09-02 | 深圳先进技术研究院 | Chimeric antigen receptor, macrophage for expressing chimeric antigen receptor and application |
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Application publication date: 20201103 |