WO2004014954A1 - Exosome containing exogenous antigen through gene transfection and method for utilizing the same - Google Patents
Exosome containing exogenous antigen through gene transfection and method for utilizing the same Download PDFInfo
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- WO2004014954A1 WO2004014954A1 PCT/KR2003/001575 KR0301575W WO2004014954A1 WO 2004014954 A1 WO2004014954 A1 WO 2004014954A1 KR 0301575 W KR0301575 W KR 0301575W WO 2004014954 A1 WO2004014954 A1 WO 2004014954A1
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- antigen
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/0005—Vertebrate antigens
- A61K39/0011—Cancer antigens
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
- A61K2039/515—Animal cells
- A61K2039/5152—Tumor cells
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
- A61K2039/515—Animal cells
- A61K2039/5156—Animal cells expressing foreign proteins
Definitions
- the present invention in general, relates to an exosome carrying an exogenous protein and use thereof. More particularly, the present invention relates to an exosome carrying an exogenous antigen, which is secreted into the extracellular space by cells transfected with a gene encoding the exogenous antigen and stably expressing the antigen, and use thereof.
- Exosomes are small cup-shaped membrane vesicles with diameters ranging from 50 to 90 nm, which are secreted by a multitude of cell types. In particular, during reticulocyte maturation, unnecessary proteins are eliminated via such exosomes. Under an electron microscope, exosomes were found not to be directly pinched off from the plasma membrane but to be derived from endosomal compartments, collectively called multivesicular bodies (MVBs), and to be released into the extracellular space. That is, upon fusion of multivesicular bodies to the plasma membrane, vesicles are released into the extracellular space, which are then called exosomes. A mechanism associated with the production of exosomes is not clearly identified.
- antigen presenting cells present antigenic peptides to MHC (major histocompatibility complex) class II molecules via intracellular membrane- bound compartments including multivesicular bodies, and thus APC-derived exosomes bear antigenic peptide-MHC class II complexes.
- exosomes as vehicles for immunogens, are able to present antigenic peptides to CD4 + T lymphocytes, resulting in the induction of specific immune responses, such as proliferation of T lymphocytes, h addition, since exosomes accumulate molecules capable of stimulating the immune response, such as MHC class I molecules or heat shock proteins (HSPs), they are effective in autoimmune disease cancer immunotherapy by increasing or reducing the immune response.
- MHC class I molecules or heat shock proteins (HSPs) molecules capable of stimulating the immune response, such as MHC class I molecules or heat shock proteins (HSPs)
- Fig. 1 is a diagram schematically showing a process of preparing exosomes carrying an exogenous protein according to the present invention
- Figs. 2a and 2b show results of SDS-PAGE, in which protein composition of an established cell line according to the present invention is compared with that of exosomes derived from the cell line;
- Figs. 3a and 3b show results of FACS analysis, in which an exogenous tumor antigen is positioned on the cell surface of a target cell line transfected with a gene encoding the exogenous antigen according to the present invention;
- Figs. 4a and 4b show results of Western blotting, in which an exogenous tumor antigen is present in exosomes isolated from a target cell line transfected with a gene encoding the exogenous antigen according to the present invention
- Fig. 5 show a result of Western blotting, in which tumor antigens are present in exosomes isolated from a human carcinoma cell line naturally carrying the tumor antigens.
- the present invention provides an exosome carrying a specific antigen by introducing a gene encoding the antigen into a target cell line by a transfection method and stably expressing the antigen therein.
- the exosome according to the present invention may be utilized to isolate proteins that are difficult to isolate while maintaining their natural states without structural damage or functional inhibition.
- the exosome may be utilized as a vehicle for immunogens when intended to induce change of the immune system employing the exogenous antigen as an immunogen.
- Exosomes originate from endosomal compartments produced during the vesicular transport from the endoplasmic reticulum (ER) to the Golgi apparatus, where antigens bind to MHC molecules, and the resulting complexes are released into the extracellular space when the endosomal compartments are fused with the plasma membrane.
- ER endoplasmic reticulum
- cell surface molecules or cell-specific antigens are produced in the ER and transferred to the cell surface via the vesicular transport pathway, resulting in exposure to the extracellular space.
- antigenic proteins newly produced in the cell may be present on the cell surface via the vesicular transport pathway, or released to the extracellular space by means of exosomes.
- the present inventors investigated that the specific antigen is stably expressed in the cells and secreted by means of exosomes.
- an exosome carrying an exogenous protein is produced by a process comprising the steps of (1) cloning a human mucl gene into a BamHI site of a pLXIN plasmid to prepare a recombinant vector, (2) introducing the recombinant vector into a packaging cell line, PA317, to produce viral particles bearing the mucl gene, (3) infecting a target cell line with the viral particles, and (4) isolating exsomes by ultra-centrifuging the tansfected cells.
- the exosomes obtained by the aforementioned process were evaluated for expression of the introduced antigen by employing a fluorescent substance or electrophoresis.
- the antigenic protein was found to be stably expressed in the target cell line, present in exosomes, and released into the extracellular space by means of exosomes.
- the exosome according to the present invention of the present invention may be applied to a wide variety of areas by employing other tumor antigens as the exogenous antigen artificially introduced into the cells.
- the exogenous antigen carried in the exosome according to the present invention has its natural structure without conformational change or degradation, which is identical to the three dimensional structure of the antigen upon exposed to the immune system in the body where tumors develop. Therefore, when intended to induce change in the immune system using an exogenous antigen, the exosome according to the present invention may be useful as a vehicle for the exogenous antigen as an immunogen.
- exosome according to the present invention may be used in isolation and purification of cell surface proteins that are difficult to be isolated and purified, macro-molecular proteins, glycosylated proteins, or complex folding-undergoing proteins, without structural damage or functional inhibition.
- Example 1 Establishment of cell line stably expressing Mucl
- Human mucl gene was cloned into a BamHI site of pLXIN (Clontech), and the resulting vector was introduced into a packaging cell line, PA 317, by means of liposomes, resulting in the production of viral particles bearing the mucl gene.
- the murine colon cancer cell line CT26 was infected with the viral particles, and selected in the presence of 1200 ⁇ g/ml of geneticin. Among the formed colonies, one clone exhibiting high expression of Mucl was determined and designated "CT26 pLXL - ucl N1010".
- each protein sample was subjected to SDS-PAGE electrophoresis.
- the cell lysate from the CT26 pLXIN-mucl N1010 clone was found to differ from the exosomes isolated from the clone in protein composition (Fig. 2a). This result indicates that the introduced mucl gene is expressed in exosomes.
- 2xl0 5 of the CT26 pLXIN-mucl N1010 cells were suspended in 0.1% BSA-containing PBS (phosphate- buffered saline) and placed on ice.
- 2 ⁇ l of a primary antibody (mouse anti-mucl IgG) was added to the cell suspension, followed by incubation for 1 hr 30 min. The cells were washed with PBS to remove unbound antibodies. Then, the cells were treated with 1 ⁇ l of a secondary antibody (FITC-tagged anti-mouse IgG) on ice for 1 hr. After washing with PBS, the cells were treated with 200 ⁇ l of a cold fixing solution (2% PFA-containing PBS). Fluorescent intensity was assayed with a Coulter FACScan flow cytometer.
- the cell surface protein Mucl was found to be present on the cell surface of the CT26 pLXIN-mucl N1010 cells prepared by introducing pLXIN-Mucl into the CT26 cells (Fig. 3a).
- Exosomes were isolated from another stably transfected clone, CT26 pLSIN-mucl N1019, prepared by introducing mucl gene into CT26 cells in Example 1.
- the isolated exosomes were subjected to SDS-PAGE, FACS analysis and Western blotting, and the results are given Figs. 2a, 3a and 4a, respectively.
- Muc 1 protein was found to be present in the surface of exosomes.
- Mucl gene was introduced into TA3HA cells according to the same procedure as in
- Example 1 thus giving a stably transfected clone, TA3HA pLXIN-mucl 615.
- Exosomes were isolated from the clone, and subjected to SDS-PAGE, FACS analysis and Western blotting, and the results are given Figs. 2b, 3b and 4b, respectively. As a result, the Mucl protein was found to be present on the surface of the exosomes.
- exosomes were isolated from the culture supernatant by ultracentrifugation, and subjected to Western blotting. The results are given in Fig. 5. As shown in Fig. 5, the tumor antigens CEA and Mucl were found to be present in the exosomes.
- exosomes were isolated from the culture supernatant by ultracentrifugation, and subjected to Western blotting. The result is given in Fig. 5. As shown in Fig. 5, the tumor antigen Mucl were found to be present in the exosomes.
- the tumor antigen Mucl was found to be secreted into the extracellular space along with exosomes.
- a desired antigen such as a vaccine to be used for induction of the immune response
- the tumor antigens CEA and Mucl were released into the extracellular space via the exosomes secreted from the carcinoma cell lines naturally expressing the tumor antigens. Based on this finding, a variety of tumor antigens can be obtained from exosomes isolated from various carcinoma cell lines, and the obtained tumor antigens can be utilized for establishment of an antigen pool or antigen bank.
- the present invention provides an exosome carrying an exogenous antigen, which is secreted by tumor cells stably transfected with a gene encoding the exogenous antigen and expressing the antigen.
- antigenic proteins as vaccines should be synthesized, isolated and then purified.
- the exosome of the present invention is effective in improving such a time-consuming and uneconomical process.
- the exosome makes it possible to isolate and purify a desired protein at its natural state, thereby reducing harmful immune responses caused by modification of the desired protein and experimental errors.
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- Animal Behavior & Ethology (AREA)
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- Pharmacology & Pharmacy (AREA)
- General Health & Medical Sciences (AREA)
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Abstract
Disclosed is an exosome carrying an exogenous antigen. The exosome of the present invention is characterized by being secreted into the extracelluar space by a target cell line transfected with a gene encoding an exogenous antigen and stably expressing the antigen, leading to the release of the antigen into the extracellular space. Therefore, the exosome facilitates the production of a variety of exogenous antigen to be used as immunogens when intended to induce change in the immune system.
Description
EXOSOME CONTAINING EXOGENOUS ANTIGEN THROUGH GENE TRANSFECTION AND METHOD FOR UTILIZING THE
SAME
Technical Field
The present invention, in general, relates to an exosome carrying an exogenous protein and use thereof. More particularly, the present invention relates to an exosome carrying an exogenous antigen, which is secreted into the extracellular space by cells transfected with a gene encoding the exogenous antigen and stably expressing the antigen, and use thereof.
Background Art
Exosomes are small cup-shaped membrane vesicles with diameters ranging from 50 to 90 nm, which are secreted by a multitude of cell types. In particular, during reticulocyte maturation, unnecessary proteins are eliminated via such exosomes. Under an electron microscope, exosomes were found not to be directly pinched off from the plasma membrane but to be derived from endosomal compartments, collectively called multivesicular bodies (MVBs), and to be released into the extracellular space. That is, upon fusion of multivesicular bodies to the plasma membrane, vesicles are released into the extracellular space, which are then called exosomes. A mechanism associated with the production of exosomes is not clearly identified. However, in addition to erythrocytes, a broad range of immune cells, including B lymphocytes, T lymphocytes, dendritic cells, platelets and macrophages, and tumor cells have been reported to secrete exosomes in living states. In particular, antigen presenting cells (APCs) present antigenic peptides to MHC (major histocompatibility complex) class II molecules via intracellular membrane-
bound compartments including multivesicular bodies, and thus APC-derived exosomes bear antigenic peptide-MHC class II complexes. Therefore, exosomes, as vehicles for immunogens, are able to present antigenic peptides to CD4+ T lymphocytes, resulting in the induction of specific immune responses, such as proliferation of T lymphocytes, h addition, since exosomes accumulate molecules capable of stimulating the immune response, such as MHC class I molecules or heat shock proteins (HSPs), they are effective in autoimmune disease cancer immunotherapy by increasing or reducing the immune response.
Disclosure of Invention
It is therefore an object of the present invention to provide an exosome carrying an exogenous antigen, which is secreted into the extracellular space by cells stably transfected with a gene encoding the exogenous antigen and expressing the antigen. It is another object of the present invention to provide use of such an exosome in purification of proteins that are difficult to be purified or as a vehicle for various immunogens.
Brief Description of the Drawings
The above and other objects, features and other advantages of the present invention will be more clearly understood from the following detailed description taken in conjunction with the accompanying drawings, in which:
Fig. 1 is a diagram schematically showing a process of preparing exosomes carrying an exogenous protein according to the present invention;
Figs. 2a and 2b show results of SDS-PAGE, in which protein composition of an established cell line according to the present invention is compared with that of exosomes derived from the cell line;
Figs. 3a and 3b show results of FACS analysis, in which an exogenous tumor antigen is positioned on the cell surface of a target cell line transfected with a gene encoding the exogenous antigen according to the present invention;
Figs. 4a and 4b show results of Western blotting, in which an exogenous tumor antigen is present in exosomes isolated from a target cell line transfected with a gene encoding the exogenous antigen according to the present invention; and Fig. 5 show a result of Western blotting, in which tumor antigens are present in exosomes isolated from a human carcinoma cell line naturally carrying the tumor antigens.
Best Mode for Carrying Out the Invention
In order to achieve the aforementioned objects, the present invention provides an exosome carrying a specific antigen by introducing a gene encoding the antigen into a target cell line by a transfection method and stably expressing the antigen therein. In addition, the exosome according to the present invention may be utilized to isolate proteins that are difficult to isolate while maintaining their natural states without structural damage or functional inhibition.
Moreover, the exosome may be utilized as a vehicle for immunogens when intended to induce change of the immune system employing the exogenous antigen as an immunogen.
Exosomes originate from endosomal compartments produced during the vesicular transport from the endoplasmic reticulum (ER) to the Golgi apparatus, where antigens bind to MHC molecules, and the resulting complexes are released into the extracellular space when the endosomal compartments are fused with the plasma membrane. On the other hand, cell surface molecules or cell-specific antigens are produced in the ER and transferred to the cell surface via the vesicular transport
pathway, resulting in exposure to the extracellular space. Similarly, antigenic proteins newly produced in the cell may be present on the cell surface via the vesicular transport pathway, or released to the extracellular space by means of exosomes. In this regard, when introducing a gene encoding a specific antigen into cells and isolating exosomes from the transfected cells, the present inventors investigated that the specific antigen is stably expressed in the cells and secreted by means of exosomes.
In accordance with the present invention, an exosome carrying an exogenous protein is produced by a process comprising the steps of (1) cloning a human mucl gene into a BamHI site of a pLXIN plasmid to prepare a recombinant vector, (2) introducing the recombinant vector into a packaging cell line, PA317, to produce viral particles bearing the mucl gene, (3) infecting a target cell line with the viral particles, and (4) isolating exsomes by ultra-centrifuging the tansfected cells. The exosomes obtained by the aforementioned process were evaluated for expression of the introduced antigen by employing a fluorescent substance or electrophoresis. As a result, the antigenic protein was found to be stably expressed in the target cell line, present in exosomes, and released into the extracellular space by means of exosomes. In addition to the Mucl tumor antigen, the exosome according to the present invention of the present invention may be applied to a wide variety of areas by employing other tumor antigens as the exogenous antigen artificially introduced into the cells.
The exogenous antigen carried in the exosome according to the present invention has its natural structure without conformational change or degradation, which is identical to the three dimensional structure of the antigen upon exposed to the immune system in the body where tumors develop. Therefore, when intended to induce
change in the immune system using an exogenous antigen, the exosome according to the present invention may be useful as a vehicle for the exogenous antigen as an immunogen.
In addition, the exosome according to the present invention may be used in isolation and purification of cell surface proteins that are difficult to be isolated and purified, macro-molecular proteins, glycosylated proteins, or complex folding-undergoing proteins, without structural damage or functional inhibition.
The present invention will be explained in more detail with reference to the following example in conjunction with the accompanying drawings. However, it will be apparent to one skilled in the art that the following examples are provided only to illustrate the present invention, and the present invention is not limited to the examples.
Example 1 (1) Establishment of cell line stably expressing Mucl
Human mucl gene was cloned into a BamHI site of pLXIN (Clontech), and the resulting vector was introduced into a packaging cell line, PA 317, by means of liposomes, resulting in the production of viral particles bearing the mucl gene.
Thereafter, the murine colon cancer cell line CT26 was infected with the viral particles, and selected in the presence of 1200 μg/ml of geneticin. Among the formed colonies, one clone exhibiting high expression of Mucl was determined and designated "CT26 pLXL - ucl N1010".
(2) Assay for Mucl expression by SDS-PAGE and enzymatic reaction lxlO7 of the transfected cells were suspended in 500 μl of RD?A buffer and incubated on ice for 10 min to allow cyto lysis, followed by centrifugation, thus giving supernatant as a protein fraction. Also, exosomes were isolated from the
transfected cells by ultracentrifugation. In the supernatant from the cell lysate and exosomes, the total protein amount was measured using a BCA protein assay kit (PIERCE). 20 μg of each protein sample was incubated with mouse anti-mucl IgG as a primary antibody and then HRP (horseradish peroxidase)-conjugated anti-mouse antibody as a secondary antibody. When a HRP substrate was added into the solution, the color developed.
Also, each protein sample was subjected to SDS-PAGE electrophoresis. The cell lysate from the CT26 pLXIN-mucl N1010 clone was found to differ from the exosomes isolated from the clone in protein composition (Fig. 2a). This result indicates that the introduced mucl gene is expressed in exosomes.
(3) Assay for Mucl expression using a fluorescent substance
In order to investigate whether Mucl is present on the cell surface, 2xl05 of the CT26 pLXIN-mucl N1010 cells were suspended in 0.1% BSA-containing PBS (phosphate- buffered saline) and placed on ice. 2 μl of a primary antibody (mouse anti-mucl IgG) was added to the cell suspension, followed by incubation for 1 hr 30 min. The cells were washed with PBS to remove unbound antibodies. Then, the cells were treated with 1 μl of a secondary antibody (FITC-tagged anti-mouse IgG) on ice for 1 hr. After washing with PBS, the cells were treated with 200 μl of a cold fixing solution (2% PFA-containing PBS). Fluorescent intensity was assayed with a Coulter FACScan flow cytometer.
As a result, the cell surface protein Mucl was found to be present on the cell surface of the CT26 pLXIN-mucl N1010 cells prepared by introducing pLXIN-Mucl into the CT26 cells (Fig. 3a).
(4) Assay for Mucl expression by Western blotting The exosomes isolated from the CT26 pLXIN-mucl N1010 cells were evaluated for carrying the Mucl protein. From the cell lysate and the isolated exosomes, 20
μg of each protein sample was subjected to SDS-PAGE, and transferred onto a nylon membrane. The membrane was sequentially incubated with antibodies specifically recognizing Mucl and HSP70 proteins. The blot was developed with chemi- luminescent reagents. As a result, Mucl protein was found to be present on the surface of the exosomes isolated from the CT26 pLXIN-mucl N1010 clone (Fig. 4a).
Example 2
Exosomes were isolated from another stably transfected clone, CT26 pLSIN-mucl N1019, prepared by introducing mucl gene into CT26 cells in Example 1. In order to investigate whether Muc 1 protein is expressed in exosomes, the isolated exosomes were subjected to SDS-PAGE, FACS analysis and Western blotting, and the results are given Figs. 2a, 3a and 4a, respectively. As shown in Figs. 2a, 3a and 4a, Muc 1 protein was found to be present in the surface of exosomes.
Example 3
Mucl gene was introduced into TA3HA cells according to the same procedure as in
Example 1, thus giving a stably transfected clone, TA3HA pLXIN-mucl 615.
Exosomes were isolated from the clone, and subjected to SDS-PAGE, FACS analysis and Western blotting, and the results are given Figs. 2b, 3b and 4b, respectively. As a result, the Mucl protein was found to be present on the surface of the exosomes.
Example 4
From another stably transfected clone, TA3HA pLXIN-mucl 617, prepared in Example 3, exosomes were isolated, and subjected to SDS-PAGE, FACS analysis and
Western blotting, and the results are given Figs. 2b, 3b and 4b, respectively. As a
. _
result, the Mucl protein was found to be present on the surface of the exosomes.
Example 5
After culturing the human gastric carcinoma cell line KATO III expressing the tumor antigens CEA and Mucl, exosomes were isolated from the culture supernatant by ultracentrifugation, and subjected to Western blotting. The results are given in Fig. 5. As shown in Fig. 5, the tumor antigens CEA and Mucl were found to be present in the exosomes.
Example 6
After culturing the human breast carcinoma cell line MCF7 expressing the tumor antigen Mucl, exosomes were isolated from the culture supernatant by ultracentrifugation, and subjected to Western blotting. The result is given in Fig. 5. As shown in Fig. 5, the tumor antigen Mucl were found to be present in the exosomes.
Upon introducing Mucl as an exogenous antigen into a target cell line in Examples 1 and 4, the tumor antigen Mucl was found to be secreted into the extracellular space along with exosomes. In this way, a desired antigen, such as a vaccine to be used for induction of the immune response, can be produced easily while maintaining its natural state. In addition, in Examples 5 and 6, the tumor antigens CEA and Mucl were released into the extracellular space via the exosomes secreted from the carcinoma cell lines naturally expressing the tumor antigens. Based on this finding, a variety of tumor antigens can be obtained from exosomes isolated from various carcinoma cell lines, and the obtained tumor antigens can be utilized for establishment of an antigen pool or antigen bank.
Industrial Applicability
As described hereinbefore, the present invention provides an exosome carrying an exogenous antigen, which is secreted by tumor cells stably transfected with a gene encoding the exogenous antigen and expressing the antigen. Upon development of tumor vaccines, antigenic proteins as vaccines should be synthesized, isolated and then purified. The exosome of the present invention is effective in improving such a time-consuming and uneconomical process.
In addition, the exosome makes it possible to isolate and purify a desired protein at its natural state, thereby reducing harmful immune responses caused by modification of the desired protein and experimental errors.
The present invention has been described in an illustrative manner, and many modifications and variations of the present invention are possible in light of the above teachings. Therefore, it is to be understood that within the scope of the appended claims, the invention may be practiced otherwise than as specifically described.
Claims
1. An exosome carrying an exogenous protein, which is secreted into the extracellular space by a target cell line transfected with a gene encoding the exogenous antigen and stably expressing the exogenous antigen.
2. Use of the exosome of claim 1 in isolation of a protein that is difficult to be isolated while maintaining its natural structure and function.
3. Use of the exosome of claim 1 as a vehicle for an immunogen when intended to use an antigen as an immunogen.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| AU2003248150A AU2003248150A1 (en) | 2002-08-13 | 2003-08-06 | Exosome containing exogenous antigen through gene transfection and method for utilizing the same |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR10-2002-0047779 | 2002-08-13 | ||
| KR10-2002-0047779A KR100519384B1 (en) | 2002-08-13 | 2002-08-13 | Manufacturing method of exosomes using gene transfection and use of the same |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2004014954A1 true WO2004014954A1 (en) | 2004-02-19 |
Family
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
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| PCT/KR2003/001575 WO2004014954A1 (en) | 2002-08-13 | 2003-08-06 | Exosome containing exogenous antigen through gene transfection and method for utilizing the same |
Country Status (3)
| Country | Link |
|---|---|
| KR (1) | KR100519384B1 (en) |
| AU (1) | AU2003248150A1 (en) |
| WO (1) | WO2004014954A1 (en) |
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1322115C (en) * | 2005-07-06 | 2007-06-20 | 清华大学 | Ectobody loaded with exogenous ligand molecule and its preparation method and application |
| GB2437727A (en) * | 2006-05-04 | 2007-11-07 | Univ Open | Aptamers against MUC1 and their uses |
| WO2011080271A2 (en) | 2009-12-28 | 2011-07-07 | Centre De Recerca En Salut Internacional De Barcelona | Exosomes derived from reticulocytes infected with plasmodium sp., method for obtaining them and uses thereof |
| WO2015085096A1 (en) * | 2013-12-04 | 2015-06-11 | Board Of Regents, The University Of Texas System | Analysis of genomic dna, rna, and proteins in exosomes for diagnosis and theranosis |
| JP2017101012A (en) * | 2015-11-30 | 2017-06-08 | 義之 小山 | Immunotherapeutic formulation |
| US10233445B2 (en) | 2009-04-17 | 2019-03-19 | Oxford University Innovation Limited | Composition for delivery of genetic material |
| US10500231B2 (en) | 2013-03-13 | 2019-12-10 | University Of Miami | Method for isolation and purification of microvesicles from cell culture supernatants and biological fluids |
| US10959952B2 (en) | 2015-06-10 | 2021-03-30 | Board Of Regents, The University Of Texas System | Use of exosomes for the treatment of disease |
| US11103586B2 (en) | 2011-12-07 | 2021-08-31 | Oxford University Innovation Limited | Exosomes for delivery of biotherapeutics |
| WO2022040516A1 (en) | 2020-08-21 | 2022-02-24 | University Of Miami | Compositions and methods of treatment using microvesicles from bone marrow-derived mesenchymal stem cells |
| WO2023205158A1 (en) | 2022-04-19 | 2023-10-26 | University Of Miami | Compositions comprising microvesicles for use in the prevention and treatment of graft versus host disease |
| US11926824B2 (en) | 2013-03-15 | 2024-03-12 | Board Of Regents, The University Of Texas System | miRNA biogenesis in exosomes for diagnosis and therapy |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3356522A4 (en) | 2016-09-30 | 2019-03-27 | Cellex Life Sciences, Incorporated | Compositions containing protein loaded exosome and methods for preparing and delivering the same |
| IL307240A (en) | 2021-03-30 | 2023-11-01 | Kainos Medicine Inc | Fas-associated factor 1 (faf1)-loaded exosomes and use thereof as anti-cancer agent |
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| WO1997005900A1 (en) * | 1995-08-03 | 1997-02-20 | Rijksuniversiteit Te Leiden | Cell derived antigen presenting vesicles |
| WO1999003499A1 (en) * | 1997-07-16 | 1999-01-28 | Institut National De La Sante Et De La Recherche Medicale | Cellular vesicle called ''exosome'', preparation and use thereof in immune stimulation |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2785543B1 (en) * | 1998-11-05 | 2003-02-28 | Inst Nat Sante Rech Med | MODIFIED EXOSOMES AND USES |
| US20040241176A1 (en) * | 2000-04-27 | 2004-12-02 | Ap Cells. Inc. | Method of producing membrane vesicles |
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2002
- 2002-08-13 KR KR10-2002-0047779A patent/KR100519384B1/en active IP Right Grant
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- 2003-08-06 WO PCT/KR2003/001575 patent/WO2004014954A1/en not_active Application Discontinuation
- 2003-08-06 AU AU2003248150A patent/AU2003248150A1/en not_active Abandoned
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| WO1997005900A1 (en) * | 1995-08-03 | 1997-02-20 | Rijksuniversiteit Te Leiden | Cell derived antigen presenting vesicles |
| WO1999003499A1 (en) * | 1997-07-16 | 1999-01-28 | Institut National De La Sante Et De La Recherche Medicale | Cellular vesicle called ''exosome'', preparation and use thereof in immune stimulation |
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| WO2023205158A1 (en) | 2022-04-19 | 2023-10-26 | University Of Miami | Compositions comprising microvesicles for use in the prevention and treatment of graft versus host disease |
Also Published As
| Publication number | Publication date |
|---|---|
| KR20040015508A (en) | 2004-02-19 |
| KR100519384B1 (en) | 2005-10-06 |
| AU2003248150A1 (en) | 2004-02-25 |
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