68 results about "Artificial antigen presenting cells" patented technology

The invention relates to novel artificial antigen presenting cells (aAPCs). The aAPC comprises at least one stimulatory ligand and at least one co-stimulatory ligand where the ligands each specifically bind with a cognate molecule on a T cell of interest, thereby mediating expansion of the T cell. The aAPC of the invention can further comprise additional molecules useful for expanding a T cell of interest. The aAPC of the invention can be used as an “off the shelf” APC that can be readily designed to expand a T cell of interest. Also, the aAPC of the invention can be used identify the stimulatory, co-stimulatory, and any other factors that mediate growth and expansion of a T cell of interest. Thus, the present invention provides powerful tools for development of novel therapeutics where activation and expansion of a T cell can provide a benefit.

The invention provides an artificial antigen presenting cell (AAPC) comprising a eukaryotic cell expressing an antigen presenting complex comprising a human leukocyte antigen (HLA) molecule of a single type, at least one exogenous accessory molecule and at least one exogenous T cell-specific epitope. Methods of use for activation of T lymphocytes are also provided.

The invention relates to novel artificial antigen presenting cells (aAPCs). The aAPC comprises at least one stimulatory ligand and at least one co-stimulatory ligand where the ligands each specifically bind with a cognate molecule on a T cell of interest, thereby mediating expansion of the T cell. The aAPC of the invention can further comprise additional molecules useful for expanding a T cell of interest. The aAPC of the invention can be used as an “off the shelf” APC that can be readily designed to expand a T cell of interest. Also, the aAPC of the invention can be used identify the stimulatory, co-stimulatory, and any other factors that mediate growth and expansion of a T cell of interest. Thus, the present invention provides powerful tools for development of novel therapeutics where activation and expansion of a T cell can provide a benefit.

T cell responses are often diminished in humans with a compromised immune system. We have developed a method to isolate, stimulate and expand naïve cytotoxic T lymphocyte precursors (CTLp) to antigen-specific effectors, capable of lysing tumor cells in vivo. This ex vivo protocol produces fully functional effectors. Artificial antigen presenting cells (AAPCs; Drosophila melanogaster) transfected with human HLA class I and defined accessory molecules, are used to stimulate CD8+ T cells from both normal donors and cancer patients. The class I molecules expressed to a high density on the surface of the Drosophila cells are empty, allowing for efficient loading of multiple peptides that results in the generation of polyclonal responses recognizing tumor cells endogenously expressing the specific peptides. The responses generated are robust, antigen-specific and reproducible if the peptide epitope is a defined immunogen. This artificial antigen expression system can be adapted to treat most cancers in a significant majority of the population.

Carbon nanotube (CNT)-based compositions for activating cellular immune responses are provided. The CNTs function as high surface area scaffolds for the attachment of T cell ligands and / or antigens. The CNT compositions function as artificial antigen-presenting cells (aAPCs) or as modular vaccines. The disclosed CNT aAPCs are efficient at activating T cells and may be used to activate T cells ex vivo or in vivo for adoptive or active immunotherapy.

Carbon nanotube (CNT)-based compositions for activating cellular immune responses are provided. The CNTs function as high surface area scaffolds for the attachment of T cell ligands and / or antigens. The CNT compositions function as artificial antigen-presenting cells (aAPCs) or as modular vaccines. The disclosed CNT aAPCs are efficient at activating T cells and may be used to activate T cells ex vivo or in vivo for adoptive or active immunotherapy.

An artificial antigen presenting cell includes a liposome having at least one recombinant soluble MHC-peptide complex incorporated therein. The artificial antigen presenting cell may also include at least one additional signal molecule incorporated therein for manipulating the intensity and quality of the immune response. The recombinant soluble MHC molecule is obtained by a method utilizing PCR amplification of gDNA or cDNA, and a tag is attached thereto for anchoring the recombinant soluble MHC molecule to the liposome.

The invention provides an in vitro amplification method for efficient and highly cytotoxic natural killer (NK) cells. Novel artificial antigen presenting cells 4-1BBL-mIL-21-aAPC, such as 4-1BBL-mIL-21-K562 cells and the like, are constructed through stably expressing 4-1BB ligands (4-1BBL) and membrane immobilized interleukin 21 (mIL-21) on the surfaces of cell membranes, and by using the novel artificial antigen presenting cells as feeder cells for amplification, the NK cells are directly amplified from peripheral blood lymphocytes. Flow cytometry, cytotoxicity test and the like suggest that the amplified cells NK have high purity and strong cytotoxicity and have obvious killing effect on tumor cells.

The invention discloses a method for in vitro induction of specific cytotoxic T lymphocytes of hepatitis B virus (HBV). The method comprises the following steps of: 1, preparing artificial antigen presenting cells (aAPC) capable of loading any polypeptides; 2, loading aAPC on epitope peptides; and 3, loading aAPC in vitro induction HBV antigen specific CTL of antigen peptides; measuring the number of obtained antigen specific CTL by a Tetramer decoration method, namely combining streptavidin marked by fluorescein and four MHC-peptide compounds marked by biotin to form the Tetramer, namely MHC-peptide tetramer; and after the MHC-peptide tetramer is combined with TCR on the antigen specific CTL, measuring by a flow cytometry. The method can be applied to the amplification of T lymphocytes resisting the HBV.

The invention provides an artificial antigen presenting cell as well as a preparation method and an application thereof. Wherein the artificial antigen presenting cell expresses membrane proteins MICA, IL-12 and CD80, the artificial antigen presenting cell is used for culturing NK cells or peripheral blood mononuclear cells containing the NK cells, the number of the NK cells can be remarkably increased, the service life of the NK cells is prolonged, the activity of the NK cells is improved, and compared with an existing artificial antigen presenting cell for expressing MICA, the artificial antigen presenting cell has a stronger effect of amplifying and activating the NK cells.

T cell responses are often diminished in humans with a compromised immune system. We have developed a method to isolate, stimulate and expand naïve cytotoxic T lymphocyte precursors (CTLp) to antigen-specific effectors, capable of lysing tumor cells in vivo. This ex vivo protocol produces fully functional effectors. Artificial antigen presenting cells (AAPCs; Drosophila melanogaster) transfected with human HLA class I and defined accessory molecules, are used to stimulate CD8+ T cells from both normal donors and cancer patients. The class I molecules expressed to a high density on the surface of the Drosophila cells are empty, allowing for efficient loading of multiple peptides that results in the generation of polyclonal responses recognizing tumor cells endogenously expressing the specific peptides. The responses generated are robust, antigen-specific and reproducible if the peptide epitope is a defined immunogen. This artificial antigen expression system can be adapted to treat most cancers in a significant majority of the population.

The present invention is directed to methods for the identification and isolation of antigen-specific T cells using a novel class of artificial antigen presenting cells. The resulting T cells may be used to produce expanded T cell populations as well as for modulating T cell responses. In general, the artificial antigen presenting cells useful in such methods are liposomes that contain MHC:peptide complexes presented on the outer surface of the liposome. Such artificial antigen presenting cells may also include accessory molecules, co-stimulatory molecules, adhesion molecules, and other molecules irrelevant to T cell binding or modulation that are used in the binding of artificial antigen presenting cells to solid support systems that may be used in the retrieval and identification of antigen-specific T cells.

The invention discloses a method for carrying out in-vitro efficient amplification on natural killer (NK) cells, and application of the method, aiming at solving the problems that the NK cells amplified by the traditional local method are small in amplification times, have cytokine dependency and cell aging, and the like. The method is characterized by comprising the following steps: 1, separating peripheral blood mononuclear cells (PBMCs) from human peripheral blood by using the traditional Ficoll-Paque density gradient centrifugation method; 2, carrying out 100 Grays irradiation treatment on artificial antigen presenting cells, and then storing by means of liquid nitrogen cryopreservation; 3, carrying out co-culture on the PBMCs and the artificial antigen presenting cells, subjected to the irradiation treatment, in a cell culture flask T75 of an RPMI 1640 medium, wherein the mass ratio of the PBMCs to the artificial antigen presenting cells is equal to 1: 2, and 50IU / mL of interleukin 2 is added into the RPMI 1640 medium; replacing the medium with a fresh medium every 2 to 3 days; 4, calculating the number of total cells in the cell culture flask T75 every 7 days, adding the same number of artificial antigen presenting cells subjected to the irradiation treatment into the cell culture flask T75 for stimulating again, and continuously culturing for 21 days to obtain the amplified NK cells. Therefore, after the method is adopted, a great deal of high-purity and high-quality NK cells can be obtained.

The present invention provides compositions and methods for immunotherapy, which include shelf-stable pharmaceutical compositions for inducing antigen-specific T cells. Such compositions are employed as components of an artificial antigen presenting cell (aAPC), to provide a patient with complexes for presentation of an antigen (e.g., a tumor antigen) and / or a T cell co-stimulatory molecule.

Carbon nanotube (CNT)-based compositions for activating cellular immune responses are provided. The CNTs function as high surface area scaffolds for the attachment of T cell ligands and / or antigens. The CNT compositions function as artificial antigen-presenting cells (aAPCs) or as modular vaccines. The disclosed CNT aAPCs are efficient at activating T cells and may be used to activate T cells ex vivo or in vivo for adoptive or active immunotherapy.

In some embodiments, compositions and methods relating to isolated artificial antigen presenting cells (aAPCs) are disclosed, including aAPCs comprising a myeloid cell transduced with one or more viral vectors, such as a MOLM-14 or a EM-3 myeloid cell, wherein the myeloid cell endogenously expresses HLA-AB / C, ICOS-L, and CD58, and wherein the one or more viral vectors comprise a nucleic acid encoding CD86 and a nucleic acid encoding 4-1BBL and / or OX40L and transduce the myeloid cell to express CD86 and 4-1BBL and / or OX40L proteins. In some embodiments, methods of expanding tumor infiltrating lymphocytes (TILs) with aAPCs and methods of treating cancers using TILs after expansion with aAPCs are also disclosed.

The invention relates to the field of immune medicine, in particular to a cell culture method which comprises the following steps: co-culturing an inactivated artificial antigen presenting cell with high expression of mIL15 and CD137L and PBMC in a culture medium, wherein the culture medium comprises a basic culture medium and additional components, the additional components comprise IL2 with the concentration of 100 IU / mL to 1200 IU / mL and zoledronic acid with the concentration of 3 mu M to 7 mu M. The ratio of NK cells to gamma delta T cells cultured by the method is high, and the cells have a more excellent anti-tumor effect.

The invention relates to CD8 cytotoxic T-lymphocyte (CTL) for treating lung cancer and a preparation method thereof. The preparation method is characterized by comprising the steps of loading antigen peptide on 668 strains of artificial antigen presenting cell (aAPC), high efficiently activating the artificial antigen presenting cell in vitro, separating CD8T cell from peripheral blood of an HLA-A2 positive tumor patient, activating the CD8T cell to be split into the CD8 cell (CTL) capable of killing the tumor cells, secreting IFN-gamma from the induced CTL, an obvious effect for killing the human tumor cell strains in vitro and self tumor cell in vivo can be realized, the CTL is returned to the patient to treat the tumor cancer or prolong the life of the patient.

The invention relates to the technical field of cell culture, particularly to an NK (natural killer) cell culture composition and a culture method. The composition comprises an artificial antigen presenting cell, IL-2, IL-15, pseudo-ginseng saponin, blood serum and a basal culture medium. The NK cell culture composition can promote mass proliferation of NK cells, improves the killing activity of the NK cells, avoids extrinsic protein and virus pollution, and reduces production cost; the obtained NK cells are relatively high in purity.

Carbon nanotube (CNT)-based compositions for activating cellular immune responses are provided. The CNTs function as high surface area scaffolds for the attachment of T cell ligands and / or antigens. The CNT compositions function as artificial antigen-presenting cells (aAPCs) or as modular vaccines. The disclosed CNT aAPCs are efficient at activating T cells and may be used to activate T cells ex vivo or in vivo for adoptive or active immunotherapy.

The present disclosure relates to artificial antigen presenting cells (aAPCs), in particular engineered erythroid cells and enucleated cells (e.g. enucleated erythroid cells and platelets), that are engineered to activate or suppress T cells.

The invention provides an artificial antigen presenting cell applied to efficiently amplifying NK and a construction method thereof. Specifically, the invention relates to fusion protein. The fusion protein is prepared from IL-15, IL-21, MICA protein and optional label sequence and / or signal peptide sequence. Experimental results show that cells of the fusion protein disclosed by the invention canvery well induce NK cells to proliferate, differentiate and activate, and the prepared NK cells has the advantages of large quantity, high purity and strong killing activity. According the artificialantigen presenting cell disclosed by the invention, genes are expressed in serial, so that 1 to 1 100% expression is achieved; meanwhile, operation procedures are simplified; furthermore, time and labor cost are greatly reduced. According the artificial antigen presenting cell disclosed by the invention, the construction method for the NK cell artificial antigen presenting cell is optimized, anda foundation is established for applying the NK cell in cancer therapy, cancer prevention and the like in later.

The invention discloses a secretory component vesica of a gene engineering-based aAPC (artificial Antigen Presenting Cell), as well as preparation and application of a T cell vaccine constructed by anexosome. By transfecting two kinds of eukaryotic expression plasmids of pcDNAHLA-A2 and pcDNACD80 and infecting two kinds of recombinant adenovirus carriers of AdVGag and AdV41BBL, a gene engineering-based K562A2 / Gag / CD80 / 41BBLaAPC is constructed; then exosomes are purified in K562A2 / Gag / CD80 / 41BBL culture supernatant through a differential superspeed centrifugal method, and electron microscopingand immunoblotting analysis can be carried out, wherein the exosomes are in co-incubation with nonspecific T cells of Con-A stimulated HLA-A2 transgenic mice through incubating, so that Gag-Texo vaccine having HLA-A-A2 limitation and Gag specificity is constructed. According to the secretory component vesica disclosed by the invention, the aAPCK562A2 / Gag / CD80 / 41BBL vesica in unlimited source andcontinuous growth is used for replacing a DC vesica in limited source and is used for preparing an HIV-1 (Human Immunodeficiency Viru I) Gag specific T cell vaccine. A novel method for preparing the Tcell vaccine by using the gene engineering-based aAPC has important influence on development of clinic therapeutic vaccines for a patient suffering HIV-1.

The invention discloses artificial antigen presenting cells and a preparation method and application thereof. According to the preparation method of the artificial antigen presenting cells, a polymermicrosphere surface topological structure morphological control method is established, an antigen presenting function of microspheres is achieved through signal molecule modification, and thus the microspheres can be used for stimulating immune cell amplification and activation. According to the artificial antigen presenting cells, adopted materials have biocompatibility and biodegradability, andthe preparation method of the microspheres with a rough surface structure is easy to operate, environmentally friendly and good in reproducibility.

The present invention relates to an exosome for stimulating T cells and the pharmaceutical use thereof. Immune exosomes secreted from artificial antigen-presenting cells which express HLA, CD32, and co-stimulatory molecules CD32, CD80, CD83, and 4-1BBL are used to stimulate naive CD8+ T cells whereby preventive and therapeutic effects on tumors, pathogen infections, or autoimmune diseases can be provided.

In some embodiments, compositions and methods relating to isolated artificial antigen presenting cells (aAPCs) are disclosed, including aAPCs comprising a myeloid cell transduced with one or more viral vectors, such as a MOLM-14 or a EM-3 myeloid cell, wherein the myeloid cell endogenously expresses HLA-A / B / C, ICOS-L, and CD58, and wherein the one or more viral vectors comprise a nucleic acid encoding CD86 and a nucleic acid encoding 4-lBBL and / or OX40L and transduce the myeloid cell to express CD86 and 4-lBBL and / or OX40L proteins. In some embodiments, methods of expanding tumor infiltratinglymphocytes (TILs) with aAPCs and methods of treating cancers using TILs after expansion with aAPCs are also disclosed.

The present invention concerns artificial antigen presenting cells (aAPCs) and methods of making and using the same, for example, to isolate, identify, and expand T cell populations specifically reactive against a disease-associated antigenic peptide, as well as to modulate responses of antigen-specific T cells both in vivo, ex vivo, and in vitro. Accordingly, the aAPCs of the invention can be used to treat conditions that would benefit from modulation of a T cell response, for example, autoimmune disorders, allergies, cancers, viral infections, and graft rejection. In certain preferred embodiments, the aAPCs are liposomes comprised of MHC:peptide complexes and accessory molecules. Other molecules, such as co-stimulatory molecules and adhesion molecules, can also be included in the compositions of the invention. In other embodiments, the aAPCs are comprised of a scaffold to which a plurality of MHC:peptide complexes and accessory molecules (as well as other molecules) can be attached at high density.

The invention relates to a method for in-vitro large-scale induction of NK cell expansion, and belongs to the technical field of biology. According to the invention, the constructed novel artificial antigen presenting cell CD86 CD64 mIL-15-aAPC is taken as an expanded feeder cell, NK cells are directly expanded from peripheral blood lymphocytes, and the NK cells expanded by the method has the advantages of good proliferation, high purity, strong cytotoxicity and obvious tumor cell killing effect. The invention solves the problems of limited number of cell expansion, low expression level of surface antigen CD3- / CD56+ and low cell killing activity faced by traditional culture methods, and can meet the clinical requirement on NK cells.

The present invention relates to method of amplifying a population of antigen-specific memory CD4+ T cells using artificial presenting cells expressing HLA class II molecules. In particular, the present invention relates to a method of amplifying a population of antigen-specific memory CD4+ T cells comprising the steps of i) providing a population of artificial antigen presenting cells consisting host cells that are genetically modified to stably express at least one MHC class II molecule along with at least one accessory molecule ii) loading the population of artificial antigen presenting cells of step i) with an amount of at least one antigen of interest and iii) coculturing the suitable population of a T cells with the population of artificial antigen presenting cells of step ii).

The present invention provides compositions and methods for immunotherapy, which include shelf-stable pharmaceutical compositions for inducing antigen-specific T cells. Such compositions are employed as components of an artificial antigen presenting cell (aAPC), to provide a patient with complexes for presentation of an antigen (e.g., a tumor antigen) and / or a T cell co-stimulatory molecule.