CN105505869A - Chimeric antigen receptor T cell for tumor stem cells - Google Patents
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Abstract
The invention provides a chimeric antigen receptor T cell for tumor stem cells. Two to three independent antigen receptors are embedded to the T cell. Each chimeric antigen receptor is formed by combining different antigen binding parts of antibodies of tumor stem cell specific markers and different functional protein, and the functional protein is selected from one or combination of two of an intracellular immunoreceptor tyrosine activation motif, a co-stimulatory factor 1 intracellular structural domain and a co-stimulatory factor 2 intracellular structural domain; an extracellular antibody part is connected with intracellular functional protein through a CD 3-zeta transmembrane domain structure. By selecting and combining the different antigen binding parts of the antibodies of the tumor stem cell specific markers (antigen) and different functional protein, the two to three independent chimeric antigen receptors are constructed, the T cell is activated only after all the two to three chimeric antigen receptors are used for recognizing antigens, and accordingly specific cytotoxicity of the tumor stem cells is achieved, and safety of the tumor stem cells are improved.
Description
Technical field
The invention belongs to biological technical field, be specifically related to a kind of Chimeric antigen receptor for tumor stem cell.
Background technology
Chimeric antigen receptor T cell (ChimericAntigenReceptorT-Cell, CAR-T) is the cell therapy technology of the treatment tumour that a kind of new development is got up.This therapeutic modality specificity is high, effective, receives the attention of medical circle.The principle of compositionality of CAR-T is, the antigen-binding portion scFv(single-chainvariablefragments of the antibody of certain tumour antigen can be identified, scFv)), with immunoreceptor tyrosine-based activation motif (immunoreceptortyrosine-basedactivationmotifs, ITAM, be generally CD3-ζ and Fc ε RI γ) born of the same parents in part in vitro coupling be a chimeric protein, by the T cell of the method transfection patient of gene transfer, it is made to express Chimeric antigen receptor (CAR).The T cell of patient, by after " recodification ", generates the specific CAR-T cell of massive tumor.
Current CAR-T lymphocyte technology has developed into the third generation, first-generation CAR is by the single-chain antibody (singlechainfragmentvariable identifying TSA, and immunoreceptor tyrosine-based activation motif (immunoreceptortyrosine-basedactivationmotifs scFv), ITAM, is generally CD3-ζ and Fc ε RI γ) composition.Early stage experiment demonstrates the feasibility of CAR-T, but first-generation CAR can only cause of short duration T cell increment and lower cytokine secretion, the anti-tumor in vivo effect that can not provide long T cell amplified signal and continue.According to the dual signal theory of T cell activation, activation and the propagation of T cell need costimulatory signal.
The intracellular signal transduction district of s-generation CAR introduces costimulating factor, be mainly CD28 molecule, enhance the multiplication capacity of T cell and the secreting function of cytokine, IL-2, INF-γ and GM-CSF increases, thus breach the immunosuppression of tumor microenvironment, extend AICD, third generation CAR is on the basis of the s-generation, add the intracellular domain of another costimulating factor, be mainly CD134(OX-40) or CD137(4-1BB) etc., be intended to the cytotoxic activity of raising T cell, proliferative and survival time, promote the release of cytokine.
The advantage that existing CAR-T exists is as follows:
1, accurate: the antigen-binding portion in chimeric antibody can efficient identification tumour antigen, precisely kills;
2, efficient: CAR-T cell can not can realize efficient identification tumour cell by antigen presenting cell approach;
Various: CAR both can utilize oncoprotein matter antigen, glycolipid class nonprotein antigen can be utilized again, expand tumour antigen target spot scope.
But there is larger shortcoming equally in existing CAR-T technology:
1, lack specific antigens: because a CAR structure only identifies a kind of antigen, and tumour specific antigen is little at present, so limit the application of CAR-T;
2, existence is manslaughtered: owing to lacking tumour specific antigen, so once use heterogenetic antigen Dispersal risk, CAR-T exists the possibility of manslaughtering after inputting human body, thus existence causes immunologic injury.
Above-mentioned shortcoming causes the clinical application of CAR-T to there is larger potential safety hazard, and its application is also restricted.
Summary of the invention
Object of the present invention is just to provide a kind of Chimeric antigen receptor T cell for tumor stem cell for solving the deficiencies in the prior art, can realize the specific tumor stem cell killed and wounded.
The object of the invention is with following technical proposals realize:
A kind of Chimeric antigen receptor T cell for tumor stem cell, this kind of T cell has been fitted together to 2 ~ 3 independently antigen receptors, each chimeric antigen receptor to be combined from different functional proteins by the antigen-binding portion of the antibody of different tumor stem cell specific marker things respectively and forms, and functional protein is selected from one or both combinations of immunoreceptor tyrosine-based activation motif in born of the same parents, costimulating factor 1 intracellular domain, costimulating factor 2 intracellular domain; Extracellular antibody moiety is connected by CD3-zeta transmembrane region with endocellular function albumen.
Described immunoreceptor tyrosine-based activation motif is CD3-ζ or Fc ε RI γ.
In described born of the same parents, costimulating factor 1 is in CD28 molecule or CD137 molecule or CD134 molecule.
In described born of the same parents, costimulating factor 2 is in the CD28 molecule different from costimulatory molecules 1 or CD137 molecule or CD134 molecule.
Described tumor stem cell is carcinoma of the pancreas stem cell, and specific marker thing is CD24+, CD44+ and CD133+.
This kind of T cell has been fitted together to 3 independently antigen receptors, and antigen receptor 1 is made up of immunoreceptor tyrosine-based activation motif in the ScFv of CD24 antibody and born of the same parents; Antigen receptor 2 is made up of costimulating factor 1 intracellular domain in the ScFv of CD44 antibody and born of the same parents; Antigen receptor 3 is made up of costimulating factor 2 intracellular domain in the ScFv of CD133 antibody and born of the same parents; Extracellular antibody moiety is connected by CD3-zeta transmembrane region with endocellular function albumen.
Described tumor stem cell is liver-cancer stem cell, and specific marker thing is CD90+ and CD44+.
This kind of T cell has been fitted together to 2 independently antigen receptors, and antigen receptor 1 is made up of immunoreceptor tyrosine-based activation motif in the ScFv of CD90 antibody and born of the same parents; Antigen receptor 2 is made up of costimulating factor 2 intracellular domain in costimulating factor 1, born of the same parents in the ScFv of CD44 antibody, born of the same parents; Extracellular antibody moiety is connected by CD3-zeta transmembrane region with endocellular function albumen.
This kind of T cell has been fitted together to 2 independently antigen receptors, and antigen receptor 1 is made up of costimulating factor 1 in immunoreceptor tyrosine-based activation motif in the ScFv of CD90 antibody and born of the same parents and born of the same parents; Antigen receptor 2 is made up of costimulating factor 2 intracellular domain in the ScFv of CD44 antibody and born of the same parents; Extracellular antibody moiety is connected by CD3-zeta transmembrane region with endocellular function albumen.
The present invention is combined from different functional proteins by the antigen-binding portion of the antibody selecting different tumor stem cell specific marker things (antigen), build independently 2-3 Chimeric antigen receptor, after only having 2-3 Chimeric antigen receptor all to identify antigen, just activated T cell, thus the specific killing realized tumor stem cell, improve its security.
Tumor stem cell has extremely strong tumorigenicity, is maintain the vitality of tumor cell group by self and unlimited increment; Motion and the ability of migrating of tumor stem cell make again the transfer of tumour cell become possibility; Tumor stem cell can be in dormant state for a long time and have several drug resistance molecule and insensitive to the extraneous chemical factors of killing tumor cell, therefore tumour often for some time recurrence after conventional tumor therapeuticing method eliminates most of Common tumors cell.At present, in neoplastic hematologic disorder, mammary cancer, cerebral tumor, prostate cancer, carcinoma of the pancreas, liver cancer, lung cancer, the existence of tumor stem cell has all been found.The CAR-T that the present invention builds can specificity tumors destroyed stem cell, eliminates the deciding factor of tumor development, invasion and attack, transfer, resistance, recurrence, can play positive promoter action to clinical treatment.
In addition, the CAR-T for tumor stem cell that this invention builds also can as the powerful of tumor research, for tumor research provides new method: (1) is studied population of cells eliminate the stem cell in tumour cell with this instrument after, to find that the presence or absence of stem cell to be born on tumor cell group the impact of depositing, developing; (2) at present also unified understanding is not reached to the identification of some tumor stem cell surface markers, can apply after method of the present invention eliminates the alternative cell of the tumor stem cell with different marker combination respectively and carry out the correlative studys such as Tumor formation, to confirm the surface marker of tumor stem cell.
Accompanying drawing explanation
Fig. 1 is that in prior art, CAR-T builds model.
Embodiment
A kind of Chimeric antigen receptor T cell for tumor stem cell, this kind of T cell has been fitted together to 2 ~ 3 independently antigen receptors, each chimeric antigen receptor to be combined from different functional proteins by the antigen-binding portion of the antibody of different tumor stem cell specific marker things respectively and forms, and functional protein is selected from one or both combinations of immunoreceptor tyrosine-based activation motif in born of the same parents, costimulating factor 1 intracellular domain, costimulating factor 2 intracellular domain; All functional proteins that 2 or 3 antigen receptors combine will comprise immunoreceptor tyrosine-based activation motif in born of the same parents, costimulating factor 1, costimulating factor 2; Extracellular antibody moiety is connected by CD3-zeta transmembrane region with endocellular function albumen.
Described immunoreceptor tyrosine-based activation motif is CD3-ζ or Fc ε RI γ; In described born of the same parents, costimulating factor 1 is CD28 molecule; In born of the same parents, costimulating factor 1 is in CD28 molecule or CD137 molecule or CD134; In born of the same parents, costimulating factor 2 is in the CD28 molecule different from costimulatory molecules 1 or CD137 molecule or CD134.
General tumor stem cell has 2-3 marker, aforesaid method is adopted to build CAR-T, but also there is minority to exist more than the tumor stem cell of 3 markers, 2-3 can be made according to the method described above and criticize the novel C AR-T low dose applications together combined for different marker, marker as large bowel cancer stem cell is CD133+24+44+ and CD166+EpCAM+, 2 batches can be made respectively for CD133+24+44+'s, and for the CAR-T low dose applications together of CD166+EpCAM+, can in conjunction with 2 kinds of CAR-T on CD133+24+44+CD166+EpCAM+ stem cell and large bowel cancer stem cell, namely the CAR-T combined is maximum, therefore stem cell is at most killed, also may exist other cells and manslaughter, but it is less to manslaughter probability, because relative to single labelled thing, the combination of these markers is that tumour cell just has, two is that often kind of CAR-T low dosage uses, even if manslaughter, the impact caused is also little.
Below for the Chimeric antigen receptor of carcinoma of the pancreas stem cell and liver-cancer stem cell.
Embodiment 1
The specific marker antigen of carcinoma of the pancreas stem cell is CD24+, CD44+ and CD133+, Chimeric antigen receptor T cell for carcinoma of the pancreas stem cell has been fitted together to 3 independently antigen receptors, and 3 antigen receptors are made up of from different functional proteins the scFv of different labelled antigen respectively; Antigen receptor 1 is made up of immunoreceptor tyrosine-based activation motif in the ScFv of CD24 antibody and born of the same parents, and can certainly be selected from CD28 molecule or CD137 molecule or any one costimulating factor intracellular domain of CD134 and forms by the ScFv of CD24 antibody; Then antigen receptor 2, antigen receptor 3 form with immunoreceptor tyrosine-based activation motif in another costimulating factor or born of the same parents respectively.
As wherein a kind of preferred scheme be: antigen receptor 1 is made up of immunoreceptor tyrosine-based activation motif CD3-ζ in the ScFv of CD24 antibody and born of the same parents; Antigen receptor 2 is made up of costimulating factor 1 intracellular domain in the ScFv of CD44 antibody and born of the same parents; Antigen receptor 3 is made up of costimulating factor 2 intracellular domain in the ScFv of CD133 antibody and born of the same parents.
Its construction process adopts fusion protein technology, and concrete steps example is as follows:
1, the Chimeric antigen receptor mark of CD24: applying gene technology, by immunoreceptor tyrosine-based activation motif encoding histone amalgamation and expression in the encoding gene of the scFv albumen of CD24 antibody and born of the same parents;
2, the Chimeric antigen receptor mark of CD44: applying gene technology, expresses the scFv encoding histone of CD44 antibody and CD28 molecule protein encode fusion;
3, the Chimeric antigen receptor mark of CD133: applying gene technology, expresses the scFv encoding histone of CD133 antibody and CD134 molecule protein encode fusion;
4, transfection: the transgenosis of these fusion roteins is entered in T cell by application rotaring dyeing technology, makes T cell express this 3 kinds of fusion roteins simultaneously.
Above-mentioned technology concrete steps are as follows:
1) preparation (for hybridoma technology) of scFV gene fragment: extract total serum IgE respectively from the hybridoma that can synthesize target antigen (CD133/24/44) antibody and synthesize cDNA(such as Promega company test kit).Take cDNA as template, to increase a complete set of VL and VH gene with the front and back primer PCR of variable region of light chain VL and variable region of heavy chain VH.By two-step approach with VL and VH fragment template, primer each other, it is single-chain antibody (scFv) gene fragment that over-lap PCR splices VL and VH at random.ScFv is connected on the expression vector Pet-28a (+) that Nde I and Xho I enzyme cut, and proceeds to intestinal bacteria Novagen company.By Ni-NTA metal affinity chromatography method purification of single stranded antibody.Utilize the single-chain antibody that ELISA and western-blot identification of escherichia coli is expressed, to determine the gene order of scFV;
2) its gene fragment is prepared according to known CD3-zeta cross-film district gene order application round pcr;
3) its gene fragment is prepared according to known CD3-ζ gene order application round pcr;
4) its gene fragment is prepared respectively according to the molecular gene sequence application round pcr of known CD28/137;
5) the gene fragment head and the tail of the gene fragment of the scFV of CD24 and CD3-zeta cross-film district's gene fragment and CD3-ζ are connected to be together in series by applying gene technology becomes the gene order of Chimeric antigen receptor 1CD133scFV-CD3-zeta-CD3-ζ;
6) in like manner, the gene fragment head and the tail of the gene fragment of the scFV of CD44 and CD3-zeta cross-film district's gene fragment and CD28 are connected to be together in series become the gene order of Chimeric antigen receptor 2CD24-CD3-zeta-CD28;
7) in like manner, the gene fragment head and the tail of the gene fragment of the scFV of CD133 and CD3-zeta cross-film district's gene fragment and CD134 are connected to be together in series become the gene order of Chimeric antigen receptor 3CD44-CD3-zeta-CD137;
8) respectively the gene order of these 3 Chimeric antigen receptor is loaded into lentiviral vectors pCMV-VSV-G(Invitrogen company);
9) transfection 293T cell is carried out by Invitrogen company Lipofectamine2000 operation instruction, after transfection, 8h is replaced by perfect medium, after cultivating 48h, collect the cell conditioned medium liquid being rich in lentiviral particle, obtain the slow virus concentrated solution of high titre after concentrated to it, measure in 293T cell and demarcate virus titer;
10) with the high titer lentivirus liquid inductance obtained dye T cell thus obtain have 3 Chimeric antigen receptor can specific recognition CD133+, the combination of CD24+, CD44+ marker the novel C AR-T cell of carcinoma of the pancreas stem cell;
11) apply: the novel C AR-T cell of acquisition venoclysis can enter human body performance antitumor action.
Embodiment 2
The specific marker antigen of carcinoma of the pancreas stem cell is CD24+, CD44+ and CD133+, and the Chimeric antigen receptor T cell for carcinoma of the pancreas stem cell has been fitted together to 3 independently antigen receptors,
Antigen receptor 1 is made up of ScFv and the CD28 molecule intracellular domain of CD24 antibody; Antigen receptor 2 is made up of ScFv and the CD137 molecule intracellular domain of CD44 antibody; Antigen receptor 3 is made up of immunoreceptor tyrosine-based activation motif CD3-ζ in the ScFv of CD133 antibody and born of the same parents; Its construction process is with embodiment 1.
Embodiment 3
The specific marker antigen of carcinoma of the pancreas stem cell is CD24+, CD44+ and CD133+, and the Chimeric antigen receptor T cell for carcinoma of the pancreas stem cell has been fitted together to 3 independently antigen receptors,
Antigen receptor 1 is made up of ScFv and the CD134 molecule intracellular domain of CD24 antibody; Antigen receptor 2 is made up of immunoreceptor tyrosine-based activation motif CD3-ζ in the ScFv of CD44 antibody and born of the same parents; Antigen receptor 3 is made up of ScFv and the CD28 molecule intracellular domain of CD133 antibody; Its construction process is with embodiment 1.
Embodiment 4
The specific marker antigen of carcinoma of the pancreas stem cell is CD24+, CD44+ and CD133+, and the Chimeric antigen receptor T cell for carcinoma of the pancreas stem cell has been fitted together to 3 independently antigen receptors,
Antigen receptor 1 is made up of ScFv and the CD137 molecule intracellular domain of CD24 antibody; Antigen receptor 2 is made up of immunity receptor in ScFv and the CD134 molecule intracellular domain born of the same parents of CD44 antibody; Antigen receptor 3 is made up of the ScFv of CD133 antibody and tyrosine activation motifs CD3-ζ; Its construction process is with embodiment 1.
Embodiment 5
The specific marker antigen of liver-cancer stem cell is CD90+ and CD44+, and the Chimeric antigen receptor T cell for liver-cancer stem cell has been fitted together to 2 independently antigen receptors, and 2 antigen receptors are made up of from different functional proteins the scFv of different labelled antigen respectively; One of them antigen receptor is made up of with wherein a kind of functional protein a kind of scFv of labelled antigen, and another one antigen receptor is made up of a kind of scFv of labelled antigen and other two kinds of functional proteins.
As one of them scheme, antigen receptor 1 is made up of immunoreceptor tyrosine-based activation motif in the ScFv of CD90 antibody and born of the same parents, and antigen receptor 2 is made up of ScFv, CD28 molecule of CD44 antibody, CD134 intracellular domain.
Above-mentioned technology concrete steps are as follows:
1) preparation (for hybridoma technology) of scFV gene fragment: extract total serum IgE respectively from the hybridoma that can synthesize target antigen (CD90/44) antibody and synthesize cDNA(Promega company test kit); Take cDNA as template, to increase a complete set of VL and VH gene with the front and back primer PCR of variable region of light chain VL and variable region of heavy chain VH.By two-step approach with VL and VH fragment template, primer each other, it is single-chain antibody (scFv) gene fragment that over-lap PCR splices VL and VH at random.ScFv is connected on the expression vector Pet-28a (+) that Nde I and Xho I enzyme cut, and proceeds to intestinal bacteria.By Ni-NTA metal affinity chromatography method purification of single stranded antibody.Utilize the single-chain antibody that ELISA and western-blot identification of escherichia coli is expressed, to determine the gene order of scFV;
2) its gene fragment is prepared according to known CD3-zeta cross-film district gene order application round pcr;
3) its gene fragment is prepared according to known CD3-ζ gene order application round pcr;
4) its gene fragment is prepared respectively according to the molecular gene sequence application round pcr of known CD28/134;
5) the gene fragment head and the tail of the gene fragment of the scFV of CD90 and CD3-zeta cross-film district's gene fragment and CD3-ζ are connected to be together in series by applying gene technology becomes the gene order of Chimeric antigen receptor 1CD90scFV-CD3-zeta-CD3-ζ;
6) in like manner, the gene fragment of the gene fragment of the scFV of CD44 and CD3-zeta cross-film district gene fragment, CD28 and CD134 head and the tail are connected to be together in series become the gene order of Chimeric antigen receptor 2CD44-CD3-zeta-CD28-CD134;
7) respectively the gene order of these 2 Chimeric antigen receptor is loaded into lentiviral vectors pCMV-VSV-G;
8) transfection 293T cell is carried out by Invitrogen company Lipofectamine2000 operation instruction, after transfection, 8h is replaced by perfect medium, after cultivating 48h, collect the cell conditioned medium liquid being rich in lentiviral particle, obtain the slow virus concentrated solution of high titre after concentrated to it, measure in 293T cell and demarcate virus titer;
9) with the high titer lentivirus liquid inductance obtained dye T cell thus obtain have 2 Chimeric antigen receptor can specific recognition CD90+, the combination of CD44+ marker the novel C AR-T cell of liver-cancer stem cell.
Embodiment 6
The specific marker antigen of liver-cancer stem cell is CD90+ and CD44+, and the Chimeric antigen receptor T cell for liver-cancer stem cell has been fitted together to 2 independently antigen receptors, and antigen receptor 1 is made up of immunoreceptor tyrosine-based activation motif in the ScFv of CD90 antibody and born of the same parents; Antigen receptor 2 is made up of ScFv, CD28 molecule of CD44 antibody, CD137 intracellular domain, and all the other are with embodiment 5.
Embodiment 7
The specific marker antigen of liver-cancer stem cell is CD90+ and CD44+, Chimeric antigen receptor T cell for liver-cancer stem cell has been fitted together to 2 independently antigen receptors, antigen receptor 1 is made up of immunoreceptor tyrosine-based activation motif, CD28 molecule intracellular domain in the ScFv of CD90 antibody and born of the same parents, antigen receptor 2 is made up of ScFv, CD137 intracellular domain of CD44 antibody, and all the other are with embodiment 5.
Embodiment 8
The specific marker antigen of liver-cancer stem cell is CD90+ and CD44+, Chimeric antigen receptor T cell for liver-cancer stem cell has been fitted together to 2 independently antigen receptors, antigen receptor 1 is made up of immunoreceptor tyrosine-based activation motif, CD134 molecule intracellular domain in the ScFv of CD90 antibody and born of the same parents, antigen receptor 2 is made up of ScFv, CD28 intracellular domain of CD44 antibody, and all the other are with embodiment 5.
Embodiment 9
The specific marker antigen of liver-cancer stem cell is CD90+ and CD44+, Chimeric antigen receptor T cell for liver-cancer stem cell has been fitted together to 2 independently antigen receptors, antigen receptor 1 is made up of immunoreceptor tyrosine-based activation motif, CD137 molecule intracellular domain in the ScFv of CD90 antibody and born of the same parents, antigen receptor 2 is made up of ScFv, CD28 intracellular domain of CD44 antibody, and all the other are with embodiment 5.
Claims (9)
1. the Chimeric antigen receptor T cell for tumor stem cell, it is characterized in that, this kind of T cell has been fitted together to 2 ~ 3 independently antigen receptors, each chimeric antigen receptor to be combined from different functional proteins by the antigen-binding portion of the antibody of different tumor stem cell specific marker things respectively and forms, and functional protein is selected from one or both combinations of immunoreceptor tyrosine-based activation motif in born of the same parents, costimulating factor 1 intracellular domain, costimulating factor 2 intracellular domain; Extracellular antibody moiety is connected by CD3-zeta transmembrane region with endocellular function albumen.
2., as claimed in claim 1 for the Chimeric antigen receptor T cell of tumor stem cell, it is characterized in that described immunoreceptor tyrosine-based activation motif is CD3-ζ or Fc ε RI γ.
3., as claimed in claim 1 for the Chimeric antigen receptor T cell of tumor stem cell, it is characterized in that costimulating factor 1 in described born of the same parents is one in CD28 molecule or CD137 molecule or CD134 molecule.
4., as claimed in claim 1 for the Chimeric antigen receptor T cell of tumor stem cell, it is characterized in that costimulating factor 2 in described born of the same parents is one in the CD28 molecule different from costimulatory molecules 1 or CD137 molecule or CD134 molecule.
5., as claimed in claim 1 for the Chimeric antigen receptor T cell of tumor stem cell, it is characterized in that described tumor stem cell is carcinoma of the pancreas stem cell, specific marker thing is CD24+, CD44+ and CD133+.
6. as claimed in claim 5 for the Chimeric antigen receptor T cell of tumor stem cell, it is characterized in that this kind of T cell has been fitted together to 3 independently antigen receptors, antigen receptor 1 is made up of immunoreceptor tyrosine-based activation motif in the ScFv of CD24 antibody and born of the same parents; Antigen receptor 2 is made up of costimulating factor 1 intracellular domain in the ScFv of CD44 antibody and born of the same parents; Antigen receptor 3 is made up of costimulating factor 2 intracellular domain in the ScFv of CD133 antibody and born of the same parents; Extracellular antibody moiety is connected by CD3-zeta transmembrane region with endocellular function albumen.
7., as claimed in claim 1 for the Chimeric antigen receptor T cell of tumor stem cell, it is characterized in that described tumor stem cell is liver-cancer stem cell, specific marker thing is CD90+ and CD44+.
8. as claimed in claim 7 for the Chimeric antigen receptor T cell of tumor stem cell, it is characterized in that this kind of T cell has been fitted together to 2 independently antigen receptors, antigen receptor 1 is made up of immunoreceptor tyrosine-based activation motif in the ScFv of CD90 antibody and born of the same parents; Antigen receptor 2 is made up of costimulating factor 2 intracellular domain in costimulating factor 1, born of the same parents in the ScFv of CD44 antibody, born of the same parents; Extracellular antibody moiety is connected by CD3-zeta transmembrane region with endocellular function albumen.
9. as claimed in claim 7 for the Chimeric antigen receptor T cell of tumor stem cell, it is characterized in that this kind of T cell has been fitted together to 2 independently antigen receptors, antigen receptor 1 is made up of costimulating factor 1 in immunoreceptor tyrosine-based activation motif in the ScFv of CD90 antibody and born of the same parents and born of the same parents; Antigen receptor 2 is made up of costimulating factor 2 intracellular domain in the ScFv of CD44 antibody and born of the same parents; Extracellular antibody moiety is connected by CD3-zeta transmembrane region with endocellular function albumen.
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| CN111378623A (en) * | 2018-12-29 | 2020-07-07 | 深圳市第三人民医院 | Targeting anti-tumor T cell and preparation method and application thereof |
| CN112673024A (en) * | 2018-06-20 | 2021-04-16 | 上海隆耀生物科技有限公司 | Chimeric antigen receptor containing co-stimulation receptor and application |
| CN114213537A (en) * | 2021-12-24 | 2022-03-22 | 北京市神经外科研究所 | CD133 antibodies, chimeric antigen receptors, and uses thereof |
| CN114316050A (en) * | 2021-12-24 | 2022-04-12 | 北京市神经外科研究所 | CD133 antibodies, chimeric antigen receptors, and uses thereof |
| CN114934071A (en) * | 2021-04-30 | 2022-08-23 | 四川大学华西医院 | CAR vector for expressing immune regulatory factor and application thereof |
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| CN108373504A (en) * | 2017-01-30 | 2018-08-07 | 亘喜生物科技（上海）有限公司 | CD24 specific antibodies and anti-CD24-CAR-T cells |
| US11697685B2 (en) | 2017-06-02 | 2023-07-11 | Asclepius (Suzhou) Technology Company Group Co., Ltd. | Chimeric antigen receptor cells targeting ROBO1, preparation method and use thereof |
| CN112673024A (en) * | 2018-06-20 | 2021-04-16 | 上海隆耀生物科技有限公司 | Chimeric antigen receptor containing co-stimulation receptor and application |
| CN111378623A (en) * | 2018-12-29 | 2020-07-07 | 深圳市第三人民医院 | Targeting anti-tumor T cell and preparation method and application thereof |
| CN111378623B (en) * | 2018-12-29 | 2023-10-20 | 深圳市第三人民医院 | Targeting anti-tumor T cell and preparation method and application thereof |
| CN115916250A (en) * | 2020-05-20 | 2023-04-04 | 美天施生物科技有限两合公司 | Compositions and methods for treating CD90 and CD326 expressing cancers |
| CN114934071A (en) * | 2021-04-30 | 2022-08-23 | 四川大学华西医院 | CAR vector for expressing immune regulatory factor and application thereof |
| CN114934071B (en) * | 2021-04-30 | 2023-10-17 | 四川大学华西医院 | CAR vector expressing immune regulation factor and application thereof |
| CN114213537A (en) * | 2021-12-24 | 2022-03-22 | 北京市神经外科研究所 | CD133 antibodies, chimeric antigen receptors, and uses thereof |
| CN114316050A (en) * | 2021-12-24 | 2022-04-12 | 北京市神经外科研究所 | CD133 antibodies, chimeric antigen receptors, and uses thereof |
| CN114213537B (en) * | 2021-12-24 | 2022-06-14 | 北京市神经外科研究所 | CD133 antibodies, chimeric antigen receptors, and uses thereof |
| CN114316050B (en) * | 2021-12-24 | 2022-11-11 | 北京市神经外科研究所 | CD133 antibodies, chimeric antigen receptors, and uses thereof |
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