CN113832103B - Preparation and application of exosome of toxoplasma infection DC - Google Patents
Preparation and application of exosome of toxoplasma infection DC Download PDFInfo
- Publication number
- CN113832103B CN113832103B CN202110932048.6A CN202110932048A CN113832103B CN 113832103 B CN113832103 B CN 113832103B CN 202110932048 A CN202110932048 A CN 202110932048A CN 113832103 B CN113832103 B CN 113832103B
- Authority
- CN
- China
- Prior art keywords
- toxoplasma
- exosomes
- cells
- exosome
- use according
- 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.)
- Active
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0634—Cells from the blood or the immune system
- C12N5/0639—Dendritic cells, e.g. Langherhans cells in the epidermis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/14—Blood; Artificial blood
- A61K35/15—Cells of the myeloid line, e.g. granulocytes, basophils, eosinophils, neutrophils, leucocytes, monocytes, macrophages or mast cells; Myeloid precursor cells; Antigen-presenting cells, e.g. dendritic cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/46—Ingredients of undetermined constitution or reaction products thereof, e.g. skin, bone, milk, cotton fibre, eggshell, oxgall or plant extracts
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/70—Undefined extracts
- C12N2500/78—Undefined extracts from protozoa
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2509/00—Methods for the dissociation of cells, e.g. specific use of enzymes
- C12N2509/10—Mechanical dissociation
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention discloses preparation and application of exocrine body of toxoplasma infected Dendritic Cells (DC); specifically, toxoplasma infection DC is utilized to obtain exosomes from DC source as a strategy for tumor treatment. The tumor volume of a mouse with colorectal cancer is reduced and the survival rate of the mouse is obviously increased by injecting the mouse into the exosomes from the toxoplasma infection DC, and the result shows that the exosomes generated by the toxoplasma infection DC have obvious tumor inhibition effect. The invention uses toxoplasma to infect DC, prepares exosomes with controllable quality and can be prepared in scale by an ultracentrifugation method to treat tumors, and provides a new method for treating tumors.
Description
Technical Field
The invention belongs to the technical field of medical biology, and particularly relates to a preparation method of exocrine for toxoplasma infection DC and application of exocrine to tumor treatment.
Background
A tumor is a disease in which cancer patients' cells are affected by genetic and environmental factors and whose proliferation is uncontrolled (Caner, A., et al Parasites and immunotherapy: immunostimulatory effect of leishmania spp.in cancer treatment (vol 110, s33, 2019). European Journal ofCancer, 2019.119:p.199-199). Cancerous changes may be caused by a variety of factors, of which the infectious parasite plays an important role in the tumor progression. Toxoplasma gondii, echinococcus granulosus and trypanosoma cruzi can relieve immunosuppression caused by tumors through Th1 type immune response and inhibit tumor growth; the cestode can inhibit inflammatory reaction generated by tumors through the generation of anti-inflammatory factors by macrophages; trypanosoma cruzi can activate host caspastes and promote apoptosis of tumor cells. Echinococcus granulosus and Trichinella spiralis can arrest cell cycle and inhibit the sustained proliferation state of tumor cells. Toxoplasma gondii, plasmodium falciparum, trypanosoma cruzi can inhibit tumor angiogenesis by inhibiting expression of VEGF (bellevilles, b.e., et al, parasites as negative regulators ofcancer. During parasitic infection, the host exhibits Th1 type immune response, achieving anti-tumor effect. However, no safe and reliable parasite strains are currently used for tumor treatment.
Dendritic Cells (DCs) are the most potent antigen presenting cells in humans. DCs can capture antigens produced during tumorigenesis and process them for presentation to T cells, thereby generating an anti-tumor T cell immune response. The mature DCs have higher MHC and costimulatory molecule expression levels and the DC-secreted exosomes carry functional MHC class I/II and T cell costimulatory molecules, and the DC-derived exosomes retain the basic immunostimulatory capacity of DCs, and the stable nature of the exosome membranes allows them to be cryopreserved for at least 6 months. Therefore, DC-derived exosomes have potential research value in Tumor therapy (Ugel, S., et al, tumor-inducedmyeloid deviation: when myeloid-derived suppressor cells meet Tumor-associpedagophages. Journal ofClinical Investigation,2015.125 (9): p.3365-3376.Tian,H.andW.Li,Dendritic cell-derived exosomes for cancer immunotherapy: hope and changes. Annals ofTranslational Medicine,2017.5 (10)).
Exosomes act as "messengers" of intercellular communication, carrying proteins and nucleic acids to target cells, exerting biological regulatory functions (Ferguson, S.W. and J.Nguyen, exosomes as therapeutics: the implications ofmolecular composition and exosomal hepatology. Journal of controlledRelease, 2016.228:p.179-190.). Can regulate the transcription and translation of genes, and plays a key role in physiological and pathological processes such as cell differentiation, development, inflammation, angiogenesis and the like. It has been found that infection of murine myoblasts (L6) exosomes by Toxoplasma gondii can inhibit tumor cell proliferation, alter the cell cycle (Baird, J.R., et al Avirulent toxoplasma gondii generates therapeutic antitumor immunityby reversing immunosuppression in the ovarian cancer microenvironment. Cancer Res,2013.73 (13): p.3842-3851.). Serum exosomes from plasmodium infected mice can inhibit angiogenesis at the tumor site (Sanders, k.l., et al, attenuated toxoplasma gondii stimulates immunity to pancreatic cancerby manipulation ofmyeloid cell potentials.cancer Immunology Research,2015.3 (8): p.891-901.). Antigen presentation of plasmodium-infected macrophage exosomes by surface MHC molecules completes activation of specific T lymphocytes (Sanders, k.l., et al Attenuated toxoplasma gondii therapy ofdisseminatedpancreatic cancer generates long-lasting immunity to pancreatic cancer.oncoimmunology,2016.5 (4): p.e 1104447.).
When the host is infected with a parasite, the host cells can transfer the "infected" information to immune cells (DC, macrophages, neutrophils, monocytes) and activate NK cells, macrophages, monocytes, T cells and B cells, exerting anti-infective effects (Wu, Z.Y., et al, extracellular vesicle-mediated communication within host-parasite interactions. Front in Immunology, 2019.9.).
The exosomes prepared by using the single cell subset infected by toxoplasma have simple components and are used as activating molecules for activating the immunosuppression of tumor-bearing mice. The exosome-derived cells can be edited later to target exosomes produced by the cells. The potential safety hazard of tumor treatment by using living insects is solved, and the effect of tumor treatment can be achieved.
Disclosure of Invention
The invention aims to overcome the safety risk of parasites in tumor treatment and provides a preparation method of toxoplasma infection DC exosomes and application thereof in tumor treatment.
The aim of the invention is realized by the following technical scheme:
the invention relates to a method for preparing exosome of toxoplasma infection DC, which comprises the following steps:
s1, separating and culturing mouse bone marrow source DC;
s2, adding the exosome medium to DC to remove the exosome medium weightToxoplasma gondii (insect strain Me 49), toxoplasma gondii and host DC cells were inoculated onto DC monolayer cells in a ratio of 1:5-5:1, 5% CO 2 Culturing in a 37 ℃ incubator for 12 hours;
s3, collecting cell culture supernatant, centrifuging 500g for 10min, and taking the supernatant; then 16500g is centrifuged for 30min; filtering the supernatant with 0.22 μm filter membrane, centrifuging 120000g of the filtered liquid for 90min, and discarding the supernatant;
s4, washing the precipitate (2 times; 120000g,90 min) with PBS, and re-suspending the precipitate with a proper amount of PBS to obtain the exosomes.
As an embodiment of the present invention, in step S1, the isolation culture includes the steps of: separating femur and tibia of a mouse, flushing bone marrow cells by a culture medium injector filled with 1640, then flushing the bone marrow cells by PBS for 2 times, and centrifuging 250g for 8min; erythrocyte lysate is lysed for 8min, counted by a blood cell counting plate, and 1640 complete medium containing GM-CSF 40ng/ml and IL-420ng/ul is added, 5% CO 2 Cell culture was carried out at 37℃for 9 days in a cell incubator to obtain bone marrow-derived DCs.
As one embodiment of the invention, the toxoplasma strain comprises Me49 and RH.
As an embodiment of the present invention, step S2 is preceded by a step of resuscitation and collection of toxoplasma gondii. The frozen toxoplasma strain is taken out from liquid nitrogen for separation culture, recovered into a Vera T25 cell culture flask containing 70%, and placed into a 37 ℃ incubator containing 5% CO2 for culture.
As an embodiment of the invention, the collecting comprises: in the recovery process, when the Vera cells release 70+/-5% of toxoplasma gondii, scraping the Vera cells infected by toxoplasma gondii by using the cells, filtering and centrifuging the cell suspension, and discarding the supernatant to obtain the toxoplasma gondii.
As an embodiment of the present invention, step S4 further comprises an identification step after the resuspension precipitation; the surface markers CD9, CD63 and TSG101 of the exosome are detected by nanoparticle tracking analysis, electron microscope observation and Western Blot.
The invention also relates to a preparation method of the toxoplasma infection DC exosome to obtain the exosome.
The invention also relates to an application of the exosome in tumor treatment.
As an embodiment of the invention, the formulation is administered intratumorally or intravenously.
Compared with the prior art, the invention has the following beneficial effects:
1) The exosomes are used for replacing toxoplasma to treat tumors, so that the potential safety hazard of treating tumors by live insects is solved;
2) By adopting a single insect strain to infect exosomes derived from a single cell strain, the problem of controllability of exosome components and functions is solved, and the method has the advantages of utilizing industrial mass production and approval of clinical medicines;
3) Exosomes prepared by a single cell subset can be edited subsequently by cells derived from the exosomes, so that exosomes generated by the cells have targeting properties;
4) The invention uses toxoplasma to infect DC, the cell source is single, the exosome component is relatively simple, and the in vitro cultured cell is beneficial to the large-scale preparation and industrial production of exosome;
5) The exosomes of the invention are derived from host cells, have good cell compatibility, can directly regulate cellular immune response, reduce toxic and side effects, and are used as natural nano-carriers for tumor treatment;
6) The invention utilizes DC source exosome infected by toxoplasma to achieve the effect of immunostimulation and tumor inhibition, and avoids the risks brought by live insect treatment.
Drawings
Other features, objects and advantages of the present invention will become more apparent upon reading of the detailed description of non-limiting embodiments, given with reference to the accompanying drawings in which:
FIG. 1 is an exosome electron microscope view; wherein, (a) toxoplasma infection DC cell-derived vesicle TeM images showing exosome morphology (scale = 100 nm); (B) Western blot analysis of vesicles derived from the DC cells infected with the beetles with anti-CD 63, CD9, TSG 101; (C) Size distribution and particle count of vesicles derived from toxoplasma infection DC 12h,24h,36h were analyzed using nanoparticle tracking;
FIG. 2 is a schematic representation of the in vivo imaging, fluorescence intensity analysis and survival curves of bone marrow derived DC exosomes of toxoplasma infected mice for treating tumor-bearing mice; wherein, (a) exosomes treat day 12 live imaging of tumor-bearing mice; 1: PBS treatment of tumor-bearing mice groups; 2: DC-derived exosomes treat tumor-bearing mice groups; 3: RH-infected DC-derived exosomes treat tumor-bearing mice groups; 4: treatment of tumor-bearing mouse exosomes with Me 49-infected DC-derived exosomes; (B) in vivo imaging mean fluorescence intensity analysis; (C) survival curve of tumor-bearing mice after exosome treatment.
Detailed Description
The present invention will be described in detail with reference to examples. The following examples will assist those skilled in the art in further understanding the present invention, but are not intended to limit the invention in any way. It should be noted that several modifications and improvements can be made by those skilled in the art without departing from the inventive concept. These are all within the scope of the present invention.
Example 1
Isolation culture of mouse bone marrow-derived dendritic cells
(1) The femur and tibia of the mice were isolated, bone marrow cells were washed with a medium syringe filled with 1640, then washed 2 times with PBS, and centrifuged at 250g for 8min. The red blood cell lysate was lysed for 8min and counted by a blood cell counting plate.
(2) 1640 complete medium (10% foetal calf serum, 1% streptomycin) containing GM-CSF 40ng/ml and IL-420ng/ul was added and incubated in a carbon dioxide cell incubator for three days.
(3) After 72h, fresh 1640 medium (containing 10% fetal bovine serum, 1% streptomycin penicillin 40ng/ml GM-CSF and 20ng/ul IL-4) was supplemented and culture continued for 72h.
Example 2
Toxoplasma culture
Frozen toxoplasma strains Me49 and RH were removed from liquid nitrogen, resuscitated into Vera T25 cell culture flasks containing 70% and placed in a 37℃incubator containing 5% CO2 for cultivation. When about 70% of toxoplasma was released, the Vera cells infected with toxoplasma were scraped off with cells, the cell suspension was then transferred to a 15ml centrifuge tube, the cell suspension was filtered 3 times with a syringe with a 25 gauge needle, after centrifugation, the supernatant was discarded and the pellet was resuspended with 10ml PBS and counted microscopically.
Example 3
Toxoplasma infection of mouse DC cells
(1) The Me49 strain was resuspended in complete medium 1640 (10% fetal bovine serum, 1% streptomycin solution) with the exosomes removed from the mouse DC cells, and the parasites were isolated from the host cells as described in 1:5 was inoculated onto DC monolayer cells, and cultured in an incubator at 37℃for 24 hours with 5% CO 2.
(3) After 12h,24h,36h, the cell culture supernatants were harvested.
Example 4
Exosome separation and identification
(1) The collected cell supernatant was centrifuged at 500g for 10min, and the supernatant was collected. Then, the mixture was centrifuged at 16500g for 30min. The precipitate was discarded and the supernatant was filtered through a 0.22 μm filter.
(2) The filtered supernatant was centrifuged at 120000g for 90min, and the pellet was washed twice with pbs. Finally, the pellet was resuspended in an appropriate amount of PBS.
(3) Exosome surface markers CD9, CD63, TSG101 were detected by Nanoparticle tracking analysis (nanomarticle TrackingAnalysis, NTA), electron microscopy, western Blot.
Example 5
Colorectal cancer tumor-bearing mouse model establishment
CT26-luc tumor cells were resuscitated and each mouse was then injected 2X 10 tumor cells by underarm injection 6 cell。
Example 6
Tumor-bearing mice for treating colorectal cancer by exosomes
Each mouse was treated 3 times (once every other day) by intratumoral injection of 50 μg per mouse.
Example 7
In vivo imaging: luciferase was injected intraperitoneally into mice and imaged in a dark box IVISTM using a chemiluminescent fluorescence imaging analysis system.
Experimental results
1 exosome isolation and identification
As shown in FIG. 1, exosomes of toxoplasma-infected DC cells were successfully obtained by ultracentrifugation. The exosome membrane structure under transmission electron microscope is cup-holder shape with diameter range of 100-200nm (figure 1A). The exosome membrane surface proteins CD63, CD9 and transmembrane protein TSG101 were detected by Westernblotting experiments and the results are shown in fig. 1B. Nanoparticle tracking analysis found that exosomes were most secreted in toxoplasma infected DC cells 12h, as shown in figure 1C.
2 exosome treatment of tumor-bearing mice
Tumor volumes were detected by in vivo imaging of mice infected with toxoplasma in DC-derived exosomes injected with colorectal cancer solid tumor mice 12, and as a result, it was found that the tumor volumes of mice treated with Me 49-infected DC-derived exosomes were significantly reduced and invisible to the naked eye (fig. 2A). The mean fluorescence intensity of Me 49-infected DC cell-derived exosomes treated mice was significantly different from that of PBS group (fig. 2B). Infection of DC cell-derived exosomes by Me49 significantly increased survival in mice (fig. 2C). The results show that the Me49 infected mouse bone marrow-derived DC cell-derived exosomes can significantly inhibit tumor growth.
It can be seen that toxoplasma Me49 infects DC cell-derived exosomes, and colorectal cancer mice are treated by tail vein injection, the tumor volume is reduced, and the survival rate of the mice is remarkably increased. The results show that exosomes produced by toxoplasma infection DC have remarkable tumor treatment effect.
In summary, the invention solves the risks associated with tumor treatment by living insects by "editing" the DC exosomes by the pathogen (toxoplasma). The method can change secretion level and exosome function of immune cell. And the exosomes can be treated by intratumoral or tail vein injection, so that the formation of tumor metastasis can be quickly and efficiently inhibited. Therefore, the invention utilizes toxoplasma to infect antigen presenting cells, the preparation quality of the exocrine thereof is controllable, and the invention can be used for large-scale preparation, thereby providing a new method for treating tumors.
The foregoing describes specific embodiments of the present invention. It is to be understood that the invention is not limited to the particular embodiments described above, and that various changes and modifications may be made by one skilled in the art within the scope of the claims without affecting the spirit of the invention.
Claims (8)
1. Use of an exosome in the preparation of a colorectal cancer treatment formulation, the exosome being prepared by a process comprising the steps of:
s1, separating and culturing mouse bone marrow source DC;
s2, adding culture medium with exosomes removed into DC to resuspend toxoplasma, inoculating toxoplasma and DC cells onto DC single-layer cells according to the ratio of 1:5-5:1, and 5% CO 2 Culturing in a 37 ℃ incubator for 12 hours;
s3, collecting cell culture supernatant, centrifuging 500g for 10min, and taking the supernatant; then 16500g is centrifuged for 30min; filtering the supernatant with a 0.22um filter membrane, centrifuging 120000g of the filtered liquid for 90min, and discarding the supernatant;
s4, washing the precipitate with PBS, and re-suspending the precipitate with a proper amount of PBS to obtain the exosomes.
2. The use according to claim 1, wherein the formulation is administered intratumorally or intravenously.
3. The use according to claim 1, wherein in step S1, the isolation culture comprises the steps of: separating femur and tibia of the mice, washing bone marrow cells with a medium injector filled with 1640, washing with PBS for 2 times, and centrifuging at 250 and g for 8min; erythrocyte lysate is lysed for 8min, counted in a blood cell counting plate, and 1640 complete medium containing GM-CSF 40ng/ml and IL-420ng/ul, 5% CO is added 2 Cell culture was carried out at 37℃for 9 days in a cell incubator to obtain bone marrow-derived DCs.
4. The use according to claim 1, wherein the toxoplasma strain comprises Me49 and RH.
5. The use according to claim 1, further comprising the step of resuscitation, collection of toxoplasma gondii before step S2.
6. The use according to claim 5, wherein the resuscitating comprises removing the cryopreserved toxoplasma strain from liquid nitrogen, resuscitating it into a flask containing 70% Vera T25 cells, and culturing it in a 37 ℃ incubator containing 5% co 2.
7. The use according to claim 5, wherein the collecting comprises: in the recovery process, when the Vera cells release 70+/-5% of toxoplasma gondii, the Vera cells infected by toxoplasma gondii are scraped off, and then the cell suspension is filtered and centrifuged, and the supernatant is discarded to obtain toxoplasma gondii.
8. The use according to claim 1, characterized in that step S4 further comprises an identification step after the resuspension precipitation; exosomes were identified by nanoparticle tracking analysis, electron microscopy, western Blot detection of exosome markers CD9, CD63 and TSG101.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110932048.6A CN113832103B (en) | 2021-08-13 | 2021-08-13 | Preparation and application of exosome of toxoplasma infection DC |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110932048.6A CN113832103B (en) | 2021-08-13 | 2021-08-13 | Preparation and application of exosome of toxoplasma infection DC |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113832103A CN113832103A (en) | 2021-12-24 |
| CN113832103B true CN113832103B (en) | 2023-08-22 |
Family
ID=78960611
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110932048.6A Active CN113832103B (en) | 2021-08-13 | 2021-08-13 | Preparation and application of exosome of toxoplasma infection DC |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113832103B (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2014179038A1 (en) * | 2013-04-29 | 2014-11-06 | Trustees Of Dartmouth College | Method for treating pancreatic cancer with toxoplasma gondii vaccine |
| CN105907715A (en) * | 2016-04-29 | 2016-08-31 | 中山大学 | Exosome of Schistosoma japonicum egg antigen (SEA) bone marrow dendritic cell origin and application thereof |
| CN107988153A (en) * | 2017-12-15 | 2018-05-04 | 英科博雅生命科技有限公司 | The method of mesenchymal stem cells derived from human umbilical blood source separation excretion body and the reagent used |
| WO2018208702A1 (en) * | 2017-05-09 | 2018-11-15 | Saint Louis University | Treatment of cancer and infectious diseases with natural killer (nk) cell-derived exosomes |
| CN109536497A (en) * | 2018-11-28 | 2019-03-29 | 同济大学 | The application of schistosoma japonicum infection and its component in human tumor prevention and treatment |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9757440B2 (en) * | 2007-10-29 | 2017-09-12 | Trustees Of Dartmouth College | Method for treating cancer with Toxoplasma gondii vaccine |
-
2021
- 2021-08-13 CN CN202110932048.6A patent/CN113832103B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2014179038A1 (en) * | 2013-04-29 | 2014-11-06 | Trustees Of Dartmouth College | Method for treating pancreatic cancer with toxoplasma gondii vaccine |
| CN105907715A (en) * | 2016-04-29 | 2016-08-31 | 中山大学 | Exosome of Schistosoma japonicum egg antigen (SEA) bone marrow dendritic cell origin and application thereof |
| WO2018208702A1 (en) * | 2017-05-09 | 2018-11-15 | Saint Louis University | Treatment of cancer and infectious diseases with natural killer (nk) cell-derived exosomes |
| CN107988153A (en) * | 2017-12-15 | 2018-05-04 | 英科博雅生命科技有限公司 | The method of mesenchymal stem cells derived from human umbilical blood source separation excretion body and the reagent used |
| CN109536497A (en) * | 2018-11-28 | 2019-03-29 | 同济大学 | The application of schistosoma japonicum infection and its component in human tumor prevention and treatment |
Non-Patent Citations (1)
| Title |
|---|
| Toxoplasma gondii GRA8-derived peptide immunotherapy improves tumor targeting of colorectal cancer;Jae-Sung Kim等;Oncotarget;第11卷(第1期);62-73 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113832103A (en) | 2021-12-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109893515B (en) | Macrophage drug-loaded microparticle preparation and preparation method thereof | |
| CA2651450C (en) | Anti-cancer medication intended for immunotherapy treatments | |
| CN110846281B (en) | Anti-tumor vaccine based on exosomes | |
| CN111265549A (en) | Surface PD-L1 molecule over-expressed mesenchymal stem cell membrane coated bionic nanoparticle and preparation and application thereof | |
| CN113456613B (en) | Construction and application of near-infrared light activated macrophage-nano prodrug targeted drug delivery system | |
| WO2012165815A2 (en) | Nano-vehicle derived from tumor tissue, and cancer vaccine using same | |
| CN107375234B (en) | Multifunctional carrier based on cell-derived vesicles in body fluid and preparation method and application thereof | |
| WO2016192451A1 (en) | Oncolytic virus formulation and preparation method thereof | |
| CN111840528A (en) | Tumor vaccine of exosome combined immune checkpoint blocking agent and preparation method thereof | |
| CN106754723B (en) | Immune cell with anti-tumor function and application thereof | |
| CN112458119A (en) | Preparation method and application of engineered bionic exosome for delivering specific protein | |
| CN109893541B (en) | Application of exosome derived from menstrual blood stem cells in preparation of medicine for treating intrauterine adhesion | |
| Nicolas-Boluda et al. | Two step promotion of a hot tumor immune environment by gold decorated iron oxide nanoflowers and light-triggered mild hyperthermia | |
| JP6235795B2 (en) | Composition for cell reprogramming | |
| CN113832103B (en) | Preparation and application of exosome of toxoplasma infection DC | |
| CN107099503A (en) | Natural killer cell high-efficient amplification in vitro cultural method with High Fragmentation power | |
| CN107693789B (en) | Compound containing cell-targeted antibody and Poly (I: C) and preparation method and application thereof | |
| CN111568931A (en) | Application of MSC (mesenchymal stem cell) in regulating and controlling human body effect CD4+ T memory cell gene expression | |
| WO2017071380A1 (en) | Tumor vaccine for use in treating liver cancer and preparation method for vaccine | |
| CN115814108A (en) | Engineered macrophage drug-loaded microparticle preparation for personalized tumor treatment and preparation method thereof | |
| KR20150020112A (en) | Mesenchymal stem cells treated metformin having immuno-modulating activity and cell therapeutic agent for preventing or treating immune disease | |
| JP2023540827A (en) | Method for producing highly pure and highly efficient naturally killed cells and their uses | |
| CN107550885B (en) | Nanoparticle carrier containing TLR3 ligand in pattern recognition receptor and preparation method and application thereof | |
| Hu et al. | Anti-tumor effects of fusion vaccine prepared by renal cell carcinoma 786-O cell line and peripheral blood dendritic cells of healthy volunteers in vitro and in human immune reconstituted SCID mice | |
| Zhang et al. | Inhibition of Cerebral Ischemia/Reperfusion Injury by MSCs-Derived Small Extracellular Vesicles in Rodent Models: A Systematic Review and Meta-Analysis |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |