CN106119193A - A kind of preparation method of the T cells with antigenic specificity having NK cell speciality concurrently - Google Patents
A kind of preparation method of the T cells with antigenic specificity having NK cell speciality concurrently Download PDFInfo
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Abstract
A kind of preparation method of the T cells with antigenic specificity having NK cell speciality concurrently.The present invention, by T cell and the antigen-presenting cell presenting specific antigen being co-cultured with certain proportion, is simultaneously introduced the IL 15 of given dose, thus activates and rapid amplifying specific T-cells.The T cell of gained had both maintained the specific killing power to the tumor cell at cell surface antigen-presenting, the most also had NK cell speciality concurrently, expresses CD56, has the strongest restrictive lethality of non-MHC, can effectively kill the tumor cell not expressing MHC.
Description
Technical field
The invention belongs to immunotherapy field, especially the cultural method of immunocyte amplification in vitro.
Background technology
As a class novel anti-cancer therapies and the traditional operation of immunotherapy, chemotherapy, radiotherapy constitute the four for the treatment of cancer
Big pillar.In immunotherapy, cellular immunotherapy embodies great development potentiality especially.Thin according to the immunity used
Born of the same parents, cellular immunotherapy can be divided into two big types.One class uses nonspecific immunocyte, such as NK cell and CIK(CD3+
CD56+) cell, both cell killing cancerous cell are that non-MHC is restrictive, the cancer cell surfaces target spot identified be not by
The antigen fragment that MHC is presented out, but the target spot that NKG2D, DNAM-1, NKp30 this kind of NK cell is identified (Pievani A,
Borleri G, et al. Blood. 2011,118 (12): 3301-3310).Another kind of therapy uses specific immunity
The T cell that cell, such as Chimeric antigen receptor are modified, the T cell that TCR modifies, or entered by TIL isolated in tumor tissues
One step expands in vitro, and directly co-cultured with antigen-presenting cell in vitro by peripheral blood amplify for specific antigen
Specific T-cells (Straathof KC, Bollard CM, et al. Blood. 2005,105 (5): 1898-1904),
The immunocyte that rear two class amplification methods are turned out is by identifying that the specific antigen being presented out by MHC kills cancerous cell.
Although utilizing specificity or nonspecific Immuno Suppressive Therapy cancer all to achieve certain curative effect (Lee JH, Lee JH, et
al. Gastroenterology. 2015,148(7):1383-1391. Chia WK, Teo M, et al. Mol Ther.
2014,22 (1): 132-139), but due to the heterogeneity of cancerous cell in cancerous tissue, the most different cancerous cell shows at its cell
The specific molecular expressed is different, individually uses non-specific or specific cellular immunotherapy to be easy to because of tumor cell
Immunologic escape and make failure in treatment (Garrido F, Aptsiauri N, et al. Curr Opin Immunol. 2016,
39:44-51).If immunocyte used has non-specific and specific killing ability simultaneously, then will effectively reduce because of
The immunologic escape that cancer cell identification scope deficiency is caused.
There is the problem that multiplication capacity is low in the T cells with antigenic specificity using current conventional method to turn out: often takes turns 7 days
With the amplification times that antigen-presenting cell co-cultures rear T cell less than 4 times of (Ramos CA, Narala N, et al. J
Immunother. 2013,36 (1): 66 76), to reach to treat the feedback cell number of required tens, need through too much
Reach the 5 common stimulations taken turns to cultivate, time-consumingly more than 1 month.On the other hand, the specific T-cells that conventional method is turned out is through long
After the In vitro culture of time, cell surface great expression PD-1.These T cell are after being fed back in the patient, and its activity can quilt
The PD-L1 of the cancer cell surfaces of patient is suppressed, thus reduces its curative effect (Wei F, Zhong S, et al. Proc Natl
Acad Sci U S A. 2013,110 (27): E2480-2489).
T cells with antigenic specificity cultural method of the present invention overcomes the limitation of conventional culture methods, cell proliferation
Speed up to every 7 days 20 times, only express extremely low PD-1 on the T cell surface turned out, but great expression CD56, and to have right simultaneously
Cancerous cell specificity and non-specific lethality.
Summary of the invention
For T cells with antigenic specificity wide not, the increasing to cancer cell identification scope overcoming current conventional method to turn out
Growing slow, the shortcoming of great expression PD-1 molecule, the present invention provides the high efficiency preparation method of a kind of T cells with antigenic specificity, institute
T cell great expression CD56 turned out, had both maintained the lethality to the cancerous cell possessing antigen presentation capability, had also had concurrently non-
The restricted lethality of MHC.
The technical scheme is that (1) is by being greatly improved the antigen-presenting cell ratio to T cells with antigenic specificity
Example, fully activates the TCR/MHC-antigenic peptides of T cell and stimulates approach altogether, thus improving T cell growth rate, promotes T cell
Express the receptor of NK cell class；(2) in culture medium, add the IL-15 of high concentration, secreted to offset after T cell is activated
The cell death of the AICD(activation induction of IL-2) effect (Waldmann T. Arthritis Res. 2002,4 (Suppl
3): S161 S167), to improve cell proliferation rate further, the expression of PD-1 is reduced.(3) control each take turns co-culture time
Between, it is to avoid the problem causing antigen-presenting cell number deficiency because incubation time is long.
The cultural method of the present invention is as it is shown in figure 1, specifically include following steps:
(1) isolated PERIPHERAL BLOOD MONONUCLEAR CELL (PBMC) is trained with the ratio of 10:1 to 40:1 altogether with antigen-presenting cell
Supporting, antigen-presenting cell can be the B lymphoblast (LCL) expressing and presenting Epstein-Barr virus antigen, it is possible to for Antigen
DC cell, gamma-delta T cell, or by CD40L(CD154) B cell that activates, these cells are all that professional antigen is presented
Cell.This co-cultures the time is 8-12 days in order to activate and be enriched with the specific T-cells for specific antigen, only
It is in order to avoid the amplification of nonspecific cell such as NK cell with a small amount of Antigen presenting cells.
(2) the above-mentioned T cells with antigenic specificity being enriched with is trained with the ratio of 1:2 to 1:6 altogether with antigen-presenting cell
Support, in culture medium, add the IL-15 of final concentration of 30 ng/ml to 120 ng/ml simultaneously.Antigen-presenting cell used can
For B lymphoblast, it is possible to for the gamma-delta T cell of Antigen, or the B cell activated by CD40L, antigen in
Delivery cell radiated by gamma-ray of available 40-80 Gy before co-culturing processes.This co-cultures the time is 6-8 days, exceedes this sky
After number, cell proliferation rate can decline.
(3) repeat step 2 and expand T cell further, until it reaches required cell number.
The invention has the beneficial effects as follows: T cells with antigenic specificity growth rate is fast, and step is simple, it is not necessary to loaded down with trivial details magnetic
Pearl purification procedures.The T cells with antigenic specificity PD-1 expression of gained is low, but great expression CD56, have special simultaneously
Property and non-specific lethality, can effectively kill by MHC at the tumor cell of cell surface antigen-presenting, it may have non-MHC
Restrictive lethality, kills the tumor cell not expressing MHC.
Accompanying drawing explanation
Fig. 1 is the cell preparation flow figure of the present invention.
Fig. 2 is the Epstein-Barr virus specific T-cells killing vigor figure to LCL cell.
Fig. 3 is the Epstein-Barr virus specific T-cells killing vigor figure to K562 cell.
Fig. 4 is the killing vigor figure of LMP2 specific T-cells PHA cell and K562 cell to having loaded LMP2.
Fig. 5 is the killing vigor figure of WT-1 specific T-cells PHA cell and K562 cell to having loaded WT-1.
Detailed description of the invention
Illustrate the present invention with example below, but the present invention is not limited thereto.Do not note in following concrete steps
The experimental technique of bright actual conditions, then according to normal condition, or the condition proposed by manufacturer is carried out.
One, the present invention is used to carry out the preparation with the Epstein-Barr virus specific T-cells of NK cell speciality.
(1) extraction 100 milliliters of peripheral bloods of volunteer, pass through density with lymphocyte separation medium Ficoll-Plaque Plus
Gradient centrifugation isolates mononuclearcell (PBMC), by the cell cryopreservation of gained after washing 2 times with PBS.
(2) 1 × 10 is taken6PERIPHERAL BLOOD MONONUCLEAR CELL ebv infection, be placed in 37 ° of C, in the CO2 incubator of 5% train
After supporting 4 weeks, obtain the B lymph matricyte system (LCL) being converted by B cell.LCL can be presented at its cell surface by MHC-I
The antigen that Epstein-Barr virus is relevant.Additionally LCL is as the B cell activated by Epstein-Barr virus, similar with the B cell activated by CD40L, all goes back
At its surface great expression CD86,4-1BBL(CD137L) etc. costimulatory molecules, become professional antigen be delivery cell (Zhu F,
Ramadan G, et al. Clin Exp Immunol. 2008,151 (2): 284-96).When LCL is thin with peripheral blood single core
When born of the same parents co-culture, wherein the specific T cell of Epstein-Barr virus will be activated amplification, through several take turns co-culture, amplification gained thin
Born of the same parents are Epstein-Barr virus specific T-cells.
(3) by after the B lymphoblast radiated by gamma-ray with 40 Gy, with PERIPHERAL BLOOD MONONUCLEAR CELL with the ratio of 1:40
Example is placed in 24 orifice plates and starts co-culturing of 12 days first round.
(4) the Epstein-Barr virus specific T-cells being enriched with the first round and B lymphoblast are in varing proportions and different
Cytokine carry out second take turns co-culture (table 1), incubation time is 7 days, renews fresh culture medium, mend simultaneously after cultivating 3 days
Fill cytokine.
The ratio of all kinds of immunocytes under the different condition of culture of table 1..
(4) by second take turns that the condition co-cultured carries out taking turns again co-culture.
(5) collect the Epstein-Barr virus specific T-cells of gained, expect that blue staining detects under the microscope and calculates institute with platform
Total cell number of results.Take a part of its cell type of cell flow cytomery.Result is as shown in table 1.Visible compare
In conventional culture methods (T cell is 4:1 with the ratio of LCL), use more LCL antigen-presenting cell (T is thin when co-culturing
Born of the same parents: LCL is 1:3), the growth rate of T cell can be improved twice, PD-1 expresses and declines simultaneously, expresses the T cell ratio of CD56
Example also increased；Further by IL-2, cytokine used being changed to IL-15, cell proliferation rate reaches every 7 days 19
Times, the T cell expressing PD-1 only has 1%, and the T cell more than 50% expresses CD56 simultaneously.
(6) separately take the T cell of a part of gained, using B lymphoblast (LCL) as target cell, test its specificity and kill
Wound activity.The DELFIA citotoxicity detection kit that detection test kit used by killing activity is produced by PerkinElmer.Knot
Fruit is as in figure 2 it is shown, the T cell using conventional culture methods and the cultural method of the present invention to be turned out has under same effect target ratio
There is similar specific killing power, when effect target ratio during at 2:1 to 40:1, to the specific cleavage rate of LCL all 40% to 90%
Between.
(7) not express the K562 cell of MHC-I as target cell, the T cell that use conventional culture methods is turned out
There is the lowest non-specific killing vigor, even if in the case of effect target is up to 40:1, the cleavage rate of the most only 20%；But utilize this
The T cell that inventive method is turned out, when effect target ratio during at 2:1 to 40:1, to the cleavage rate of K562 between 50% to 90% (as
Shown in Fig. 3).This shows that the cell turned out by the present invention has the strongest restricted lethality of non-MHC simultaneously.
(8) in order to further determine that co-culture time T cell and antigen-presenting cell optimal proportion, will be at different cell ratios
The T cell turned out under example is with the ratio of the double positive cell of flow cytomery CD3+CD56+ therein.Result such as table 2
Shown in, along with the increase of LCL cell concentration, the T cell ratio of CD56+ is also being continuously increased, when ratio is 1:2 close to 50%, and 1:
To the highest by 53% when 4, it is further added by 1:6 also remaining unchanged close to 50%, if but continuing to increase LCL cell number, the then cell gathered in the crops
In containing the dead LCL cell of higher proportion, so the ratio of T cell and antigen-presenting cell should be selected between 1:2 to 1:6.
The impact of the ratio of the different T cell of table 2. and the antigen-presenting cell CD3+CD56+ cell concentration on being turned out.
| T cell: LCL | 4:1 | 2:1 | 1:1 | 1:2 | 1:4 | 1:6 |
| CD3+ CD56+(%) | 15.2 | 19.3 | 28.7 | 48.9 | 53.1 | 47.4 |
(9) in order to determine co-culture time IL-15 optium concentration, by the cell turned out under variable concentrations with streaming
Cell instrument detects the ratio of the double positive cell of CD3+CD56+ therein.Result is as shown in table 3, along with the increase of IL-15 concentration,
The T cell ratio of CD56+ is also continuously increased, and when concentration 30 ng/ml close to 50%, reaches the highest by 51.3% during 90 ng/ml,
Concentration is further added by 120 ng/ml the most no longer having increase, it is seen that the optium concentration of IL-15 should be selected in 30 more than ng/ml, in conjunction with
Cost consideration, does not the most require more than 120 ng/ml.
The impact of the different IL-15 concentration CD3+CD56+ cell concentration on being turned out of table 3..
| IL-15 (ng/ml) | 7 | 15 | 30 | 60 | 90 | 120 |
| CD3+ CD56+(%) | 18.2 | 26.3 | 46.7 | 49.9 | 51.3 | 50.8 |
(10) often take turns, in order to determine, the Best Times co-cultured, T cell be co-cultured with the ratio of 1:3 with LCL, used
The concentration of IL-15 is 40 ng/ml, every the cell number that sampling detection in 2 days is amplified.Result is as shown in table 4, co-cultures
First 6 days cell proliferation rapid, but amplification rate is decreased obviously after the 8th day.Therefore often take turns the time co-cultured and should control at 6-8
My god, hereafter should add more LCL and start co-culturing of a new round.
Table 4. difference co-cultures the T cell amplification times after the time.
| Co-culture the time (my god) | 2 | 4 | 6 | 8 | 10 |
| Cells expanded (compared with initial T cell number) | 2.7 | 7.8 | 16.3 | 24.2 | 26.9 |
Two, the present invention is used to carry out the preparation with the LMP2 specific T-cells of NK cell speciality.
The purpose of this experiment is to verify that the method for the present invention can be thin for antigen load with the B cell activated by CD40L
Born of the same parents cultivate has specific T-cells that high CD56 expresses, for LMP2 virus antigen.
(1) extraction 100 milliliters of peripheral bloods of volunteer, pass through density with lymphocyte separation medium Ficoll-Plaque Plus
Gradient centrifugation isolates mononuclearcell (PBMC), by standby for the cell cryopreservation of gained after washing 2 times with PBS.
(2) obtain stable expression CD40L(CD154 with slow-virus transfection U251 human glioma cells system) U251-CD40L
Cell.Add prior to cultivating in 24 orifice plates after U251-CD40L covers whole bottom, then the radiated by gamma-ray with 80 Gy to cell
PBMC to 1 × 105/ ml co-cultures, and adds the ciclosporin A (Cyclosporin A) of final concentration of 1 ug/ml in culture medium
And 100 IL-4 of U/ml, it being placed in 37 ° of C, cultivate 1 week in the CO2 incubator of 5%, the cell of gained is again with 4 × 105/ ml's is dense
Degree repeatedly co-cultures with irradiated U251-CD40L cell.B cell in PBMC can be activated by CD40L and expand in a large number
Increasing, the B cell (B blast) being activated, in addition to expressing MHC-I, goes back the costimulatory molecules such as great expression CD86,4-1BBL, becomes
For professional antigen be delivery cell (Kondo E, Gryschok L, et al. Clin Exp Immunol. 2009,155 (2):
249-256), can effective stimulus specific amplification T cell after Antigen.
(3) antigen that cell is loaded is that the peptide library covering Epstein-Barr virus LMP2 albumen (is purchased from JPT Technology's
PepMix, each polypeptide length is 15 aminoacid, and wherein 11 aminoacid are overlapping with adjacent polypeptide).By peptide library with 10
The final concentration of ng/ml adds the B cell being activated can complete antigen load in 1 hour.
(4) it is placed in 24 orifice plates with the ratio of 1:20 opens having loaded the B cell of LMP2 and PERIPHERAL BLOOD MONONUCLEAR CELL
Co-culturing of 10 days first round of beginning.
(5) T cell being enriched with the first round and the B cell having loaded LMP2 carry out the second common training taken turns with 1:3 ratio
Supporting, add IL-15 to final concentration of 40 ng/ml in culture medium, incubation time is 7 days, renews fresh culture medium after cultivating 3 days, with
Time supplement IL-15.
(6) by second take turns that the condition co-cultured carries out taking turns again co-culture, the specific T of LMP2 collecting gained is thin
Born of the same parents.Expect that blue staining detects and calculate the total cell number gathered in the crops under the microscope with platform.Take a part of cell streaming thin
Born of the same parents' instrument detects its cell type, and testing result shows that wherein the double positive cell proportion of CD3+CD56+ is 46%, expresses PD-1's
Cell is 2.3%.
(7) preparing PHA cell (PHA blast) for the detection of Execution, concrete preparation process is as follows: take 5 ×
105The PBMC of/ml, in culture medium, addition phytohaemagglutinin (PHA) is to 5 μ g/ml, adds IL-2 to 100 U/ after cultivating 2 days
Ml, divides the cell expanded to 24 new orifice plates after 2 days, repeats the stimulating course of above PHA and IL-2, collect gained
PHA cell, a part loads LMP2 antigen as stated above, and another part does not has Antigen as a control group.
(8) the LMP2 specific T-cells of gained is taken, respectively to have loaded the PHA cell of LMP2, not had the PHA of Antigen
And K562 cell is as target cell, test is to they killing activities.Detection test kit used by killing activity is
The DELFIA citotoxicity detection kit that PerkinElmer is produced.Result as shown in Figure 4, uses the present invention to turn out
LMP2 specific T-cells does not has lethality to the PHA cell not having Antigen, but has the strongest to the PHA having loaded antigen
Specific killing power, when effect target ratio is during at 2:1 to 40:1, and specific cleavage rate is between 40% to 90%.The T cell cultivated
The K562 cell not expressing MHC-I is also had the strongest lethality, when effect target ratio during at 2:1 to 40:1, cleavage rate 40% to
Between 80%.This shows using the B cell activated by CD40L as antigen-presenting cell, the LMP2 specificity turned out by the present invention
T cell has the strongest specificity and the restrictive non-specific lethality of non-MHC simultaneously.
Three, the present invention is used to carry out the preparation with the WT-1 specific T-cells of NK cell speciality.
The purpose of this experiment is to verify that the method for the present invention can be with gamma-delta T cell for antigen load cell
Cultivate and there is specific T-cells that high CD56 expresses, for WT-1 cancer antigen.
(1) extraction 100 milliliters of peripheral bloods of volunteer, pass through density with lymphocyte separation medium Ficoll-Plaque Plus
Gradient centrifugation isolates mononuclearcell (PBMC), by standby for the cell cryopreservation of gained after washing 2 times with PBS.
(2) 5 × 10 are taken7PERIPHERAL BLOOD MONONUCLEAR CELL, in culture medium, add the zoledronic acid of final concentration of 5 uM
And the IL-2 of 200 U/ml (Zometa), it is placed in 37 ° of C, cultivates 2 weeks in the CO2 incubator of 5%, the gamma-delta in PMBC
T cell is activated and expands in a large number, and the gamma-delta T cell being activated, in addition to expressing MHC-I, goes back great expression
The costimulatory moleculeses such as CD86,4-1BBL, becoming professional antigen is delivery cell (Maniar A, Zhang X, et al. Blood.
2010,116 (10): 1726-1733), can effective stimulus specific amplification T cell after Antigen.
(3) antigen that cell is loaded is that the peptide library covering cancer specific protein WT-1 (is purchased from JPT
The PepMix of Technology, each polypeptide length is 15 aminoacid, and wherein 11 aminoacid are overlapping with adjacent polypeptide).
With the gamma-delta T cell that the final concentration addition of 10 ng/ml is activated, peptide library can be completed antigen for 1 hour bear
Carry.
(4) the gamma-delta T cell having loaded WT-1 is put with the ratio of 1:20 with PERIPHERAL BLOOD MONONUCLEAR CELL
In 24 orifice plates, start co-culturing of 10 days first round, be simultaneously introduced IL-15 to 40 ng/ml.
(5) T cell being enriched with the first round and the gamma-delta T cell having loaded WT-1 are carried out with 1:3 ratio
Second take turns co-culture, add IL-15 in culture medium to final concentration of 40 ng/ml, incubation time is 7 days, renews after cultivating 3 days
Fresh culture medium, supplements IL-15 simultaneously.
(6) by second take turns that the condition co-cultured carries out taking turns again co-culture, the specific T of WT-1 collecting gained is thin
Born of the same parents.Expect that blue staining detects and calculate the total cell number gathered in the crops under the microscope with platform.Take a part of cell streaming thin
Born of the same parents' instrument detects its cell type, and testing result shows that wherein the double positive cell proportion of CD3+CD56+ is 58%, expresses PD-1's
Cell proportion is 1%.
(7) detection for Execution of the PHA cell is prepared.Specifically comprise the following steps that and take 5 × 105The PBMC of/ml, in
In culture medium, addition phytohaemagglutinin (PHA) is to 5 μ g/ml, adds IL-2 to 100 U/ml, will expand after 2 days after cultivating 2 days
The cell increased divides to 24 new orifice plates, repeats the stimulating course of above PHA and IL-2, collects the PHA cell of gained, a part
Load WT-1 antigen as stated above, another part does not has Antigen as a control group.
(8) the WT-1 specific T-cells of gained is taken, respectively to have loaded the PHA cell of WT-1, not had the PHA of Antigen
And K562 cell is as target cell, test is to they killing activities.Detection test kit used by killing activity is
The DELFIA citotoxicity detection kit that PerkinElmer is produced.Result is as it is shown in figure 5, use the WT-that the present invention turns out
1 specific T-cells does not has lethality to the PHA cell not having Antigen, but has the PHA having loaded antigen the strongest special
Property lethality, when effect target ratio during at 2:1 to 40:1, specific cleavage rate is between 30% to 75%.The T cell cultivated is not to
The K562 cell expressing MHC-I also has the strongest lethality, when effect target ratio during at 2:1 to 40:1, cleavage rate 40% to 80% it
Between.This shows using gamma-delta T cell as antigen-presenting cell, the WT-1 specific T-cells turned out by the present invention
There is the strongest specificity and the restrictive non-specific lethality of non-MHC simultaneously.
Claims (9)
1. prepare a cultural method for T cells with antigenic specificity, by by T cell and the antigen presentation presenting specific antigen
Cell repeatedly co-cultures, thus activates and expand for the specific T-cells being rendered antigen, it is characterized in that:
When () co-cultures beginning a T cell and professional antigen be the ratio of delivery cell be 1:2 to 1:6.
2. (b) adds the IL-15 that concentration is 30 ng/ml to 120 ng/ml when co-culturing.
3. (c) co-cultures the time every time is 6 to 8 days.
Cultural method the most according to claim 1, is characterized in that: T cell used for by PERIPHERAL BLOOD MONONUCLEAR CELL with
The antigen-presenting cell of antigen-presenting co-cultures 8-12 days gained with the ratio of 10:1 to 40:1.
Cultural method the most according to claim 1, is characterized in that: antigen-presenting cell used is for being converted institute by Epstein-Barr virus
The B lymphoblast obtained.
Cultural method the most according to claim 1, is characterized in that: antigen-presenting cell used is the B activated by CD40L
Cell.
Cultural method the most according to claim 1, is characterized in that: antigen-presenting cell used is gamma-delta T
Cell.
Cultural method the most according to claim 1, is characterized in that: the antigen being rendered is virus antigen.
Cultural method the most according to claim 1, is characterized in that: the antigen being rendered is cancer cell specific antigen.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112831468A (en) * | 2019-11-22 | 2021-05-25 | 路春光 | TCM/TSCM cell for improving TIL cell induction and application |
| CN114949190A (en) * | 2022-04-29 | 2022-08-30 | 苏州易慕峰生物科技有限公司 | Application of antigen presenting cell and CAR-T cell combination in anti-tumor |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104411819A (en) * | 2012-06-11 | 2015-03-11 | 威尔森沃尔夫制造公司 | Improved methods of cell culture for adoptive cell therapy |
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| CN104411819A (en) * | 2012-06-11 | 2015-03-11 | 威尔森沃尔夫制造公司 | Improved methods of cell culture for adoptive cell therapy |
Non-Patent Citations (3)
| Title |
|---|
| PATRICK J. HANLEY等: "expanding cytotoxic T lymphocytes from umbilical cord blood that target cytomegalovirus, epstein—barr virus, and adenovirus", 《JOURNAL OF VISUALIZED EXPERIMENTS》 * |
| VERONICA E.GARCIA等: "IL-15 enhances the response of human γδT cells to nonpetide microbial antigens", 《THE JOURNAL OF IMMUNOLOGY》 * |
| 薛莉: "人白细胞介素15真核基因表达及对小鼠肺腺癌的基因治疗", 《中国优秀硕博士学位论文全文数据库（博士）医药卫生科技辑》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112831468A (en) * | 2019-11-22 | 2021-05-25 | 路春光 | TCM/TSCM cell for improving TIL cell induction and application |
| CN114949190A (en) * | 2022-04-29 | 2022-08-30 | 苏州易慕峰生物科技有限公司 | Application of antigen presenting cell and CAR-T cell combination in anti-tumor |
| CN114949190B (en) * | 2022-04-29 | 2024-04-30 | 苏州易慕峰生物科技有限公司 | Application of antigen presenting cells and CAR-T cell combination in anti-tumor |
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