CN103468640A - Method of improving killing effect of CIK cells on cancer cells - Google Patents
Method of improving killing effect of CIK cells on cancer cells Download PDFInfo
- Publication number
- CN103468640A CN103468640A CN2013104543568A CN201310454356A CN103468640A CN 103468640 A CN103468640 A CN 103468640A CN 2013104543568 A CN2013104543568 A CN 2013104543568A CN 201310454356 A CN201310454356 A CN 201310454356A CN 103468640 A CN103468640 A CN 103468640A
- Authority
- CN
- China
- Prior art keywords
- cell
- interleukin
- cik
- add
- cells
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Abstract
The invention relates to the field of medical treatment, in particular to a method of improving the killing effect of CIK cells on cancer cells. The method of improving the killing effect of the CIK cells on the cancer cells specifically comprises the following steps: in the process of inducing the CIK cells, a CD3 monoclonal antibody, interleukin-1, interleukin-2, interferon IFN-gamma and interleukin-24 are added. By means of the method of improving the killing effect of the CIK cells on the cancer cells, the effect-target ratio is increased, and the killing activity to tumor cells is promoted.
Description
Technical field
The present invention relates to medical field, in particular to improving the method for CIK cell to the lethal effect of cancer cells.
Background technology
Cytokine induced kill cell (cytokine-induced killer cells, CIK cell) is human peripheral blood single nucleus cell to be induced jointly with cytokine profiles in vitro and a group foreign cell that obtains.Large quantity research show the CIK cell be a kind of novel, efficient, there is the restricted immune effector cell of non-major histocompatibility complex (MHC) that wide spectrum kills tumor activity, manifest huge using value in immunotherapy of tumors, have than LAK, CTL, multiplication capacity and cytotoxicity that TIL is stronger, and restricted without MHC, thereby become the study hotspot of nearly ten years tumor biotherapies.
The activating technology of CIK cell adopts the CD3 monoclonal antibody usually, interleukin 1, interleukin II and Interferon, rabbit INF-γ are activated and are increased, the cell amplification effect is not satisfied, a lot of units are short for reduction cultivation cost causes the cell cultures time in addition, report that individually incubation time is only 2-3 days, and the cell quantity therefore obtained is limited, (CIK: cancer cells) ratio is low, is difficult to meet the needs of clinical treatment for the effect target.
Summary of the invention
The object of the present invention is to provide the method for a kind of CIK of raising cell to the lethal effect of cancer cells, to address the above problem.
The method of the raising CIK cell provided in an embodiment of the present invention to the lethal effect of cancer cells specifically comprises the following steps:
In the process of CIK cell induction, add CD3 monoclonal antibody, interleukin 1, interleukin II, Interferon, rabbit IFN-γ and interleukin II 4.
In the method for raising CIK cell provided by the invention to the lethal effect of cancer cells, CD3 monoclonal antibody wherein and IFN-γ are the essential of each scheme, and the CD3 monoclonal antibody plays the mitogen activity effect, can be crosslinked with T cell surface CD3, induce its activation; Interferon, rabbit IFN-γ can induce the synthetic of the factor such as interleukin 1.The main difference of different schemes is the difference of interleukin, at present, the investigator is in the activity of being devoted to improve the CIK killing tumor cell, what have adds interleukin 7 or interleukin 12 or Interleukin-15 etc., also has applying gene transformation and monoclonal antibody to carry out the report of cell modification.In the present invention, added interleukin II 4 in the process of inducing at CIK, traditional method has been improved.
Interleukin II 4(interleukin-24, IL-24) be field of immunology research in recent years than multiple cytokine, there is the various biological activity, there is the effect of significant inhibition tumour, can optionally suppress the growth of kinds of tumors, inducing apoptosis of tumour cell.
Raising CIK cell provided by the invention has promoted the quick increment of CIK cell to the method for the lethal effect of cancer cells, generally in two weeks left and right cell total amounts of cultivation, can reach 1x10
10above, for clinical treatment provides more sufficient effector cell, improved effect target ratio, promoted the killing activity to tumour cell.
Promoted the increment of cell simultaneously due to interleukin II 4, can reduce to a certain extent incubation time, relatively reduced patient's waiting time, this measure provides experimental basis for the anticancer clinical application of CIK cell.
The accompanying drawing explanation
Fig. 1. for the use of observing under light microscopic raising provided by the invention CIK cell to the method for the lethal effect of cancer cells at cell induction observation figure A under CIK cell mirror after six days;
Fig. 2. for the use of observing under light microscopic raising provided by the invention CIK cell to the method for the lethal effect of cancer cells at cell induction after a couple of days, the observation figure B under CIK cell mirror;
Fig. 3 A is for being used the cultivation group that adds IL-24 to induce the variation diagram of the CIK cell appreciation rate of CIK cell;
Fig. 3 B induces the variation diagram of the CIK cell appreciation rate of CIK cell for the cultivation group that does not add IL-24;
Fig. 4. while inducing 20 days, do not add the cell phenotype CD3 of the cultivation group of IL-24
+, CD56
+detected result;
Fig. 5. while inducing 20 days, add the cell phenotype CD3 of the cultivation group of IL-24
+, CD56
+detected result;
Fig. 6. the observation figure of optical microphotograph Microscopic observation CIK cell killing morphological observation;
Fig. 7. the aspect graph of the normal tumour cell of seeing under scanning electron microscope;
Fig. 8. under scanning electron microscope, cell is seen kills and wounds under state the CIK cell around HepG2, the aspect graph contacted with tumour cell;
Fig. 9. part CIK cell enters tumour cell by the aspect graph of the microvillus of tumour cell parcel;
Figure 10. the aspect graph of the downright bad disintegration of Partial tumors cell of seeing under scanning electron microscope;
Figure 11. the aspect graph that the tumour cell of seeing under transmission electron microscope is surrounded by a lot of active lymphocytes;
Figure 12. the tumour cell of the apoptosis of seeing under transmission electron microscope, the Partial tumors cell has entered the aspect graph in tumour cell;
Figure 13. the downright bad tumour cell showed increased of the cultivation group that adds IL-24 of seeing under transmission electron microscope, the aspect graph of the complete pyknosis of core;
Figure 14. be full of a large amount of fat in the kytoplasm of seeing under transmission electron microscope and dripped particle, mitochondrial cristae swelling, vacuolar degeneration, the complete pyknosis of parts of fine karyon, the nuclear membrane structure is unclear, the consoluet aspect graph of membrane structure in cell.
Embodiment
Below by specific embodiment, also by reference to the accompanying drawings the present invention is described in further detail.
Embodiment 1:
Improve the method for CIK cell to the lethal effect of cancer cells, specifically comprise the following steps:
In the process of CIK cell induction, add CD3 monoclonal antibody, interleukin 1, interleukin II, Interferon, rabbit IFN-γ and interleukin II 4.
In the method for raising CIK cell provided by the invention to the lethal effect of cancer cells, CD3 monoclonal antibody wherein and IFN-γ are the essential of each scheme, and the CD3 monoclonal antibody plays the mitogen activity effect, can be crosslinked with T cell surface CD3, induce its activation; Interferon, rabbit IFN-γ can induce the synthetic of the factor such as interleukin 1.The main difference of different schemes is the difference of interleukin, at present, the investigator is in the activity of being devoted to improve the CIK killing tumor cell, what have adds interleukin 7 or interleukin 12 or Interleukin-15 etc., also has applying gene transformation and monoclonal antibody to carry out the report of cell modification.In the present invention, added interleukin II 4 in the process of inducing at CIK, traditional method has been improved.
Interleukin II 4(interleukin-24, IL-24) be field of immunology research in recent years than multiple cytokine, there is the various biological activity, there is the effect of significant inhibition tumour, can optionally suppress the growth of kinds of tumors, inducing apoptosis of tumour cell.
Raising CIK cell provided by the invention has promoted the quick increment of CIK cell to the method for the lethal effect of cancer cells, generally in two weeks left and right cell total amounts of cultivation, can reach 1x10
10above, for clinical treatment provides more sufficient effector cell, improved effect target ratio, promoted the killing activity to tumour cell.
Promoted the increment of cell simultaneously due to interleukin II 4, can reduce to a certain extent incubation time, relatively reduced patient's waiting time, this measure provides experimental basis for the anticancer clinical application of CIK cell.
Embodiment 2:
Based on above-described embodiment, the present invention has also done following improvement:
In the process of CIK cell induction, while adding CD3 monoclonal antibody, interleukin 1, interleukin II and Interferon, rabbit INF-γ, specifically add in the following order:
Add the CD3 monoclonal antibody in the CIK cell;
Add Interferon, rabbit IFN-γ and interleukin II 4;
Add again interleukin 1, interleukin II after 24 hours.
Wherein, after 24 hours, add again interleukin 1, also comprise after the step of interleukin II:
While going down to posterity amplification, again supplement interleukin II, Interferon, rabbit INF-γ and interleukin II 4 at every turn.
Described going down to posterity at least comprises 5 times.
Wherein, interleukin II 4 add-ons the number according to the amount of CIK cell the number add, induce effect in order to improve, needing to keep the concentration of interleukin II 4 in nutrient solution added is 1000IU/mL, the concentration of CIK cell remains on 10 usually
6iU/mL, the add-on of remaining CD3 monoclonal antibody, interleukin 1, interleukin II and Interferon, rabbit INF-γ is identical with common usage quantity.
Use the Experimental Comparison of experimental group provided by the invention and traditional control group as follows:
Materials and methods
The preservation 1.1 material HepG2 tumor cell line You Zhe goes down to posterity at center, Interferon, rabbit (IFN-γ), interleukin 1 (IL-1) are U.S. Peprotech company product, CD3 is Beijing Tian-Guang Biotechnology Co., Ltd. Is's product, interleukin II (IL-2) is purchased from Jiangsu spun gold profit pharmaceutcal corporation, Ltd, interleukin II 4(IL-24) purchased from R&D systems.
The mouse-anti people CD3 of Percp combination, the mouse-anti people CD4 of FITC combination, the mouse-anti people CD8 of PE combination, the mouse-anti people CD56 of PE combination, CD3 is the U.S. company BD product, foetal calf serum is the biological responsibility of Hangzhou folium ilicis chinensis company limited product, the GT-T551 substratum is purchased from Japanese Bao doctor's thing company, healthy human peripheral blood You Zhe center volunteer provides, lymphocyte separation medium is Tianjin Hao ocean biological products science and technology limited Company product, MTT is Beijing Suo Laibao Science and Technology Ltd. product, flow cytometer is the U.S. company BD product, enzyme mark spectrophotometric is counted BioRAU, 450 types, Americanized.The fluorescence inverted microscope is Japanese OLYMPUS company product.
1.2 method
1.2.1
Experimental group: the induction method that adds the CIK cell of IL-24 cultivation group specifically comprises the steps:
1. the machine of singly adopting is adopted donor autologous peripheral blood mononuclearcell, about 40-50mL/ bag, anti-freezing;
2. the human lymphocyte parting liquid is put to 50mL centrifuge tube bottom, add the equivalent blood sample of patient, wherein the volume ratio that adds that adds volume and blood sample of patient of human lymphocyte parting liquid is 1:1, and human lymphocyte is when separating, centrifugal speed is 2000rpm, centrifugation time 20min;
3. carefully draw white circular layer single core lymphocyte, put in the 50mL centrifuge tube, add appropriate nutrient solution and microbiotic;
4. add nutrient solution 0.5mL, piping and druming mixes, and regulates cell concn;
5. by the adding in the cell bottle of coated CD3 monoclonal antibody 100ng/mL, add self-inactivating blood plasma 10% simultaneously, then add Interferon, rabbit INF-γ 1000U/mL and IL-24,1000IU/mL, cultivate 24h;
6. observation of cell, add IL-1 α 100IU/mL, adds the GT-T551 substratum 10mL containing IL-21000IU/mL, IL-24,1000IU/mL;
7.24h backsight cell amplification situation is suitably diluted cell with substratum, is incubated in culturing bottle or culture bag according to the concrete quantity of cell;
8. within later every 2-3 days, go down to posterity amplification cultivation once.Supplement IL-2, INF-γ and IL-24,1000IU/mL simultaneously.
Control group: the induction method that does not add the CIK cell of IL-24 cultivation group specifically comprises the steps:
1. the machine of singly adopting is adopted donor autologous peripheral blood mononuclearcell, about 40-50mL/ bag, anti-freezing;
2. the human lymphocyte parting liquid is put to 50mL centrifuge tube bottom, add the equivalent blood sample of patient, wherein the volume ratio that adds that adds volume and blood sample of patient of human lymphocyte parting liquid is 1:1, and human lymphocyte is when separating, centrifugal speed is 2000rpm, centrifugation time 20min;
3. carefully draw white circular layer single core lymphocyte, put in the 50mL centrifuge tube, add appropriate nutrient solution and microbiotic;
4. add nutrient solution 0.5mL, piping and druming mixes, and regulates cell concn;
5. by the adding in the cell bottle of coated CD3 monoclonal antibody 100ng/mL, add self-inactivating blood plasma 10% simultaneously, then add Interferon, rabbit INF-γ 1000U/mL to cultivate 24h;
6. observation of cell, add IL-1 α 100U/mL, adds the GT-T551 substratum 10mL containing IL-21000IU/mL;
7.24h backsight cell amplification situation is suitably diluted cell with substratum, is incubated in culturing bottle or culture bag according to the concrete quantity of cell;
8. within later every 2-3 days, go down to posterity amplification cultivation once.Supplement IL-2 and INF-γ simultaneously.
1.2.2CIK the mensuration of cell-proliferation activity
The CIK cell that two kinds of methods are cultivated is with 5 * 10
7/ mL is placed in culturing bottle, and every three days supplementary fresh mediums and cytokines are also used the blue counting cells sum of placenta calculating mean value.
1.2.3CIK the mensuration of cytotoxic activity
The tumour cell of taking the logarithm vegetative period, adjust cell concn and be respectively 1 * 10
5/ mL, 5 * 10
4/ mL, 2.5 * 10
4/ mL, every hole 100ul is laid in 96 orifice plates, establishes four multiple holes, puts 37 ℃, 5%CO for every group
2cultivate in incubator, after 24h, effector cell CIK concentration is adjusted into to 1 * 10
6/ mL and target cell concentration are respectively 10:1,20:1,40:1 adds in 96 orifice plates, separately establish independent target cell group (target cell+nutrient solution) and the negative control group of individual effect groups of cells (effector cell+nutrient solution), after effect 48h, every hole adds the MTT20ul/ hole, continue to cultivate 4h～6h, then turnover panel abandoning supernatant, every hole adds 100ul DMSO, the concussion several minutes, use microplate reader (wavelength 570nm) mensuration absorbance A value after precipitating whole the dissolving, calculating kill rate (%)=1-[A(effect+target)-the A(effect)]/the A(target) * 100%.
1.2.4CIK cell phenotype is measured
Get 2 * 10
6/ mL cell is in the streaming dedicated pipe, and corresponding antibodies mixes lucifuge and places 20min, with PBS, washes the centrifugal 5min of twice, 1500rpm, and the machine for the treatment of after 1% formaldehyde is fixing is measured.
Two groups of cells (do not add the IL-24 control group and add the IL-24 experimental group and detected respectively CD3 in 8,15,20 days
+, CD4
+, CD8
+, CD3
+, CD56
+the cytolemma sign.
1.2.5 under light microscopic, cellular form is observed
In Induction Process every day observation of cell metamorphosis under light microscopic, with the tumour cell co-cultivation, observe both modal difference.
1.2.6 scanning electron microscope and transmission electron microscope sample prepare this laboratory method of reference.
Statistical procedures adopts the SPSS10.0 data analysis software, and all data mean with mean ± SD, adopt variance analysis and paired t-test statistical analysis, and P<0.05 is for there being statistical significance.
2 test-results
2.1 the variation of cellular form in Induction Process
The mononuclearcell size of just having extracted is than homogeneous, refractivity is more consistent, and in the active cells through cultivating after inducing in 3 days, the part cell obviously increases, visible a lot of microcolony starts to generate, and karyon density is strengthened, and the cell atypia is more obvious, visible volume particle in nucleus is large granular lymphocyte (LGL).
As shown in Figure 1, observation of cell colony showed increased and increase in the 6th day, can see white spot colony (seeing Figure 1A) under naked eyes, as shown in Figure 2, under mirror, large colony becomes black group, does not see cell, but has a lot of cells to swim out of around colony, the full refractivity of cell is good, size and form different (seeing Figure 1B).
A: induce CIK cell Microscopic observation after six days, cell is atypia, and colony formation (* 400) is arranged;
B: cell rapid amplifying after a couple of days, be the laminated type growth, transparence weakens, visible several large colonies (* 200) under light microscopic.
As shown in Fig. 3 A and Fig. 3 B, the number of days of X-coordinate for cultivating, the sum that ordinate zou is the CIK cell.Add IL-24 cultivation group and do not add IL-24 cultivation group and all when cultivating 3 days, start propagation, enter the fast breeding phase when cultivating 15 days, within 21 days, breed and be tending towards slowly entering plateau later, add IL-24 cultivation group propagation slightly faster than not adding IL-24 cultivation group (Fig. 2), learn by statistics the paired t-test analysis, t>3.055, p<0.05, there were significant differences for both
Results suggest IL-24 can promote CIK cell proliferation to a certain extent.
2.3CIK the killing activity of cell
The inhibiting rate (mean ± SD) of two kinds of CIK cells of table 1 to tumour cell
Add as known from Table 1 IL-24 cultivation group the effect target when identical kill rate all apparently higher than other group, the variance analysis statistical study with do not add IL-24 cultivation group and compare and significant difference is all arranged (p<0.05), do not add the different effect of IL-24 cultivation group target than between inhibiting rate there were significant differences (p<0.05), than between there is no notable difference but add the different effect of IL-24 cultivation group targets, can find out that adding the IL-24 cultivation organizes each different effect targets and compare tumour cells and all have very high inhibiting rate.
2.4CIK cell phenotype is measured
As shown in Figure 4 and Figure 5, along with the prolongation CD3 of induction time
+and CD56
+two positive cell percentages significantly raise, and adding IL-24 cultivation group and not adding IL-24 cultivation group is respectively (6.52 ± 0.7) %, (15.8 ± 0.4) %, (19.85 ± 1.3) % and (4.78 ± 0.9) %, (14.58 ± 1.6) %, (20.18 ± 1.3) % when inducing 8,15,20 days.Between two groups of different induction times, cell phenotype does not have notable difference.
Two groups of cell phenotype CD3 while inducing 20 days
+, CD56
+detected result (transverse axis is CD3+, and the longitudinal axis is CD56
+, fourth quadrant is two positive phenotypes CD3
+, CD56
+cell).
2.5CIK cell killing morphological observation
2.5.1 the optical microphotograph Microscopic observation, as shown in Figure 6,
CIK cell killing HepG2 tumour cell
The CIK cells contacting, around tumour cell, in figure, maxicell is tumour cell, minicell is CIK cell (* 400).
2.5.2 under scanning electron microscope, cellular form is observed
As shown in Figure 7 to 10, observe and can see that normal tumour cell is irregularly shaped under scanning electron microscope, surface abundant microvilli and elongated (Fig. 4 A), kill and wound under state the CIK cell around HepG2, the CIK cell contacts with tumour cell, and tumour cell obviously diminishes (Fig. 4 B), two kinds of cell close contacts, some CIK cells enter tumour cell by the microvillus of tumour cell parcel (Fig. 4 C), downright bad disintegration of some tumour cells (Fig. 4 D).
A: the normal about 10um(of tumour cell diameter * 3700);
The B:CIK cell is around HepG2, tumour cell obviously diminish (* 4000);
C: two kinds of cell close contacts, some CIK cells are by tumour cell parcel (* 2700);
D: the downright bad disintegration of tumour cell, periphery is CIK cell (* 1700).
2.5.3 under transmission electron microscope, cellular form is observed, as shown in Figure 11 to 14,
Figure A, can see under transmission electron microscope that tumour cell is by a lot of active lymphocytic encirclements;
Figure B, can see apoptotic tumor cell, and some tumour cells have entered in tumour cell;
Figure C, add IL-24 and organize downright bad tumour cell showed increased, the complete pyknosis of core;
Figure D, be full of a large amount of fat and dripped particle in kytoplasm, mitochondrial cristae swelling, and vacuolar degeneration, the complete pyknosis of parts of fine karyon, the nuclear membrane structure is unclear, and in cell, membrane structure is dissolved fully.
The A:CIK cell surrounds tumour cell (* 2500);
The B:CIK cell enters tumour cell (* 5000);
C: pyknosis, nuclear membrane is unclear, and single-layer membrane structure mitochondrial cristae fracture (* 6000) is (IL-24);
D: the double membrane structure meromixis with together with, thicken, nuclear fusion forms core prominent (* 10000) (IL-24).
Thoroughly cure tumour with method Chang Buneng such as traditional operation, chemotherapy, radiotherapies.As one of method of immunotherapy, the killer cell that the application cell factor is induced can external a large amount of amplifications because of it, cytotoxic activity is strong, can kill and wound autologous and allosome kinds of tumor cells, CIK surface existing T cell sign (CD3), the surface marker (CD56) of NK cell is also arranged, thereby have T lymphocyte anti-tumor activity and the restricted characteristics of killing knurl of the non-MHC of NK cell concurrently.Can continue division and proliferation after feeding back in body and, without the maintaining of the foreign cell factor, few side effects, shown good development prospect.
Usually, the incubation time of CIK cell is longer, and patient's rear general need to cultivate of taking a blood sample can reach 5 * 10 in 2-3 week
9the quantity clinical treatment CIK cell quantity standard of appointment (the Chinese immunology can) of left and right, so a longer waiting time of needs of patients.
Induce the scheme of CIK cell to have multiple, wherein CD3 monoclonal antibody and IFN-γ are the essential of each scheme, and the former plays the mitogen activity effect, can be crosslinked with T cell surface CD3, induce its activation, and the latter can induce the synthetic of the factor such as IL-1.The main difference of different schemes is the difference of interleukin, and at present, the investigator is in the activity of being devoted to improve the CIK killing tumor cell, and what have adds IL-7 or IL-12 or IL-15 etc., also has applying gene transformation and monoclonal antibody to carry out the report of cell modification.
In the present invention, in the process of inducing at CIK, added IL-24, traditional method is improved, innovative approach is as follows.
1. cultivation composition: in the CD3 monoclonal antibody, IL-1, IL-2, add Celeuk 4 in INF-γ culture system, strengthens the activation value-added effect to the CIK cell.
2. upper selection of joining day: when first the cultivation, IL-24 and INF-γ add simultaneously, add IL-1 after 24 hours again, IL-2, and while then going down to posterity, (at least 5 times) also corresponding interpolation IL-24 plays a role it from start to finish in culture system at every turn.
Aforesaid method has promoted the quick increment of CIK cell, generally in two weeks left and right cell total amounts of cultivation, can reach 1x10
10above, for clinical treatment provides more sufficient effector cell, improved effect target ratio, promoted the killing activity to tumour cell.
IL-24 has promoted the increment of cell, can reduce to a certain extent incubation time, has relatively reduced patient's waiting time,
This measure provides experimental basis for the anticancer clinical application of CIK cell.
The foregoing is only the preferred embodiments of the present invention, be not limited to the present invention, for a person skilled in the art, the present invention can have various modifications and variations.Within the spirit and principles in the present invention all, any modification of doing, be equal to replacement, improvement etc., within all should being included in protection scope of the present invention.
Claims (4)
1. improve the method for CIK cell to the lethal effect of cancer cells, it is characterized in that, specifically comprise the following steps:
In the process of CIK cell induction, add CD3 monoclonal antibody, interleukin 1, interleukin II, Interferon, rabbit IFN-γ and interleukin II 4.
2. the method for raising CIK cell according to claim 1 to the lethal effect of cancer cells, is characterized in that,
In the process of described CIK cell induction, while adding CD3 monoclonal antibody, interleukin 1, interleukin II and Interferon, rabbit INF-γ, specifically comprise following addition sequence:
Add the CD3 monoclonal antibody in the CIK cell;
Add Interferon, rabbit IFN-γ and interleukin II 4;
Add again interleukin 1, interleukin II after 24 hours.
3. the method for raising CIK cell according to claim 2 to the lethal effect of cancer cells, is characterized in that,
Add again interleukin 1 after described 24 hours, also comprise after the step of interleukin II:
While going down to posterity amplification, supplement interleukin II, Interferon, rabbit INF-γ and interleukin II 4 at every turn.
4. the method for raising CIK cell according to claim 3 to the lethal effect of cancer cells, is characterized in that,
Described going down to posterity at least comprises 5 times.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310454356.8A CN103468640B (en) | 2013-09-27 | 2013-09-27 | Method of improving killing effect of CIK cells on cancer cells |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310454356.8A CN103468640B (en) | 2013-09-27 | 2013-09-27 | Method of improving killing effect of CIK cells on cancer cells |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103468640A true CN103468640A (en) | 2013-12-25 |
| CN103468640B CN103468640B (en) | 2015-02-18 |
Family
ID=49793682
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310454356.8A Expired - Fee Related CN103468640B (en) | 2013-09-27 | 2013-09-27 | Method of improving killing effect of CIK cells on cancer cells |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103468640B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105154399A (en) * | 2015-02-17 | 2015-12-16 | 上海市胸科医院 | Method for cultivating lymph-node autologous CIK (cytokine-induced killer) cells and application of lymph-node autologous CIK cells |
| CN107418933A (en) * | 2017-08-25 | 2017-12-01 | 河南省银丰生物工程技术有限公司 | A kind of external evoked amplification method of cik immunocytes |
-
2013
- 2013-09-27 CN CN201310454356.8A patent/CN103468640B/en not_active Expired - Fee Related
Non-Patent Citations (2)
| Title |
|---|
| 袁玉涛 等: "IL-24对CIK细胞体内杀伤肿瘤细胞活性的影响", 《实用肿瘤学杂志》 * |
| 郭云泉 等: "CIK细胞的体外扩增能力及杀瘤效应", 《新疆医科大学学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105154399A (en) * | 2015-02-17 | 2015-12-16 | 上海市胸科医院 | Method for cultivating lymph-node autologous CIK (cytokine-induced killer) cells and application of lymph-node autologous CIK cells |
| CN107418933A (en) * | 2017-08-25 | 2017-12-01 | 河南省银丰生物工程技术有限公司 | A kind of external evoked amplification method of cik immunocytes |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103468640B (en) | 2015-02-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Ni et al. | Adoptively transferred natural killer cells maintain long-term antitumor activity by epigenetic imprinting and CD4+ T cell help | |
| CN102618498B (en) | Preparation method for HLA-A0201 limited antigen specificity CTL (cytotoxic T lymphocyte) | |
| CN103923880B (en) | The CTL preparation method of a kind of high efficiently multiplying, target killing tumour | |
| CN103756963A (en) | Method used for in vitro proliferation of NK cells | |
| Cany et al. | Combined IL-15 and IL-12 drives the generation of CD34+-derived natural killer cells with superior maturation and alloreactivity potential following adoptive transfer | |
| CN104593324A (en) | Natural killer cell culture medium and natural killer cell amplification culture method | |
| CN103849599A (en) | Culture medium efficiently amplifying autologous NK cells and cultural method | |
| CN105567634A (en) | Culture medium and method for NK cell expansion in vitro | |
| CN105062968A (en) | DC-CIK (Dendritic cell-Cytokine Induced Killer) cell culture reagent and culture method thereof | |
| CN102676454B (en) | Preparation method for CIK (cytokine induced killer) cell of umbilical cord blood source | |
| CN102268405A (en) | Method for auto NK (Natural Killer) cell in-vitro activation and amplification culture and special culture medium thereof | |
| Natarajan et al. | Ibrutinib enhances IL-17 response by modulating the function of bone marrow derived dendritic cells | |
| CN103756961A (en) | Method used for in vitro induced amplification of NKT cells | |
| CN104711225A (en) | In-vitro preparation method of NK cells | |
| Yang et al. | Discovery of a novel natural killer cell line with distinct immunostimulatory and proliferative potential as an alternative platform for cancer immunotherapy | |
| CN115558641B (en) | High-purity effector immune cell population, culture method, reagent composition and application thereof | |
| CN102988415B (en) | Natural killer cells (NK) prepared by industrializing human allogeneic nucleated cells and injection | |
| CN116333986A (en) | Culture method for exosome activated NK cells | |
| CN104694472A (en) | Method for amplifying and cryopreserving natural killer cells | |
| CN115651903A (en) | High-lethality immune cell population and culture method, reagent composition and application thereof | |
| CN105238753A (en) | Kit used for preparing CTL and application thereof | |
| CN102719400B (en) | Preparation method of HLA-A0201 restrictive anti-carcinoembryonic antigen (CEA) antigenic specificity cytotoxic T lymphocyte (CTL) | |
| CN103740643B (en) | The external evoked cultural method of the restricted killer T cell of a kind of MHC | |
| CN103468640B (en) | Method of improving killing effect of CIK cells on cancer cells | |
| Mintz et al. | Exploiting Human CD34+ Stem Cell–conditioned Medium for Tissue Repair |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20150218 Termination date: 20180927 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |