Fibroblasts and microvesicles thereof for reduction of toxicity associated with cancer immunotherapy

Abstract

To provide novel strategies to control cytokine release syndromes, and especially cytokine storms.SOLUTION: Embodiments of the disclosure include methods and compositions related to treatment and prevention of an excess of cytokines in an individual using fibroblasts or fibroblast-derived microvesicles. In particular embodiments, there are methods and compositions for treating and preventing toxicities in an individual that may be the result of cytokine release syndrome. In specific cases, an individual is treated for cytokine release syndrome with fibroblasts having one or more specific markers.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] (CROSS REFERENCE TO RELATED APPLICATIONS) This application is a continuation of U.S. Provisional Patent Application No. 62 / 793,545, filed January 17, 2019. This provisional application claims priority to US Provisional Patent Application No. 2005 / 0133636, filed on May 13, 2005, which is incorporated herein by reference in its entirety.

[0002] Embodiments of the present disclosure relate to the fields of cell biology, molecular biology, cell therapy and medicine, including cancer. At least [Background technology]

[0003] Malignant tumors have become one of the major diseases threatening human health and survival. Surgery, radiation, Conventional cancer treatments, such as radiation therapy and chemotherapy, involve either directly removing the tumor or irradiating the tumor cells. They require an external force, including being killed by radiation or chemotherapy drugs. and radiation therapy usually cannot solve problems such as tumor metastasis and recurrence. These treatments always result in severe toxic effects and damage to normal cells. Therapy and chemotherapy target the immune system, particularly N, which plays a key role in the body's anti-tumor defenses. Impairs K-cell and T-cell mediated immunity. Interestingly, in some cases In this study, depression and fatigue associated with conventional cancer treatment were improved by immunoscoping following administration of a psychotropic antidepressant. Neuropathic role in immune suppression, as evidenced by patients recovering from hypercalcaemia. There has been increasing interest in immunotherapy-based biological treatments. This approach has the potential to specifically kill tumor cells while sparing non-malignant cells. It has the advantage of not causing the horrific side effects usually associated with conventional cancer treatments. Unfortunately, despite recent successes with immunotherapy, checkpoint inhibitors Even with cellular approaches such as chimeric antigen receptor (CAR)-T cell approaches, However, a new type of adverse effect to immunotherapy was observed. In contrast to the side effects of chemotherapy or radiation therapy, which occur due to cell killing, immunotherapy The side effects are due to excessive cytokine production, which has a clinical name called "cytokine release syndrome." Related.

[0004] Cytokine release syndrome is an adverse effect of various immunotherapies and in certain cases An example of cytokine release syndrome is the chimeric antigen receptor agonist (CAAR) syndrome. This occurs in some patients who have received T cell immunotherapy, such as CAR-T cells. When these cells are activated, they cause a systemic inflammatory response and are released into the bloodstream. In response to the immune system, cytokines are released rapidly and in large quantities, resulting in serious symptoms such as severe hypotension, high fever, and tremors. In more severe cases of cytokine release syndrome, the interaction between cytokines and white blood cells This creates a positive feedback loop that leads to extremely high levels of cytokines. Cytokine storm (also known as cytokine cascade or hypercytokinemia) ) patients experience adverse reactions, such as cardiac dysfunction, adult respiratory distress syndrome, neurotoxicity, renal potentially life-threatening complications, such as hepatic and / or liver failure, pulmonary edema, and disseminated intravascular coagulation. This may lead to potential complications.

[0005] One devastating example of a cytokine storm is the induction of cytokine storms by antibodies that bind to the CD28 receptor on T cells. This was observed in a Phase I trial of the anti-cancer drug TGN1412, which was administered intravenously once. After 90 minutes of exposure, all six volunteers reported headache, muscle pain, nausea, diarrhea, erythema, vasodilation and A systemic inflammatory response characterized by rapid induction of proinflammatory cytokines accompanied by hypotension. Within 12 to 16 hours of injection, the volunteers developed pulmonary infiltrates and damage, renal failure, and The patient was in critical condition with severe lymphocyte and monocyte hyperplasia and disseminated intravascular coagulation. The unexpected depletion occurred within 24 hours of infusion. All six patients were in the intensive care unit of a public hospital. The patient was transferred to a hospital for treatment at the ER, where he was given intensive cardiopulmonary support (including dialysis), high-dose methylprednisolone, and The patient received chemotherapy and anti-interleukin-2 receptor antagonist antibody. Two patients developed prolonged cardiovascular shock and acute respiratory distress syndrome over 8 and 16 days. Intensive organ support was required throughout the course of the disease. Evidence of multiple cytokine release syndromes Despite these complications, all six patients survived the crisis.

[0006] The usual treatment for cytokine release syndrome is biologic therapy such as corticosteroids and anti-IL6 therapy. Treatment consists of chemotherapy, chemotherapy alone ... may affect the activity and / or proliferation of A cytokine storm involves a very large number of cytokines, Because of the limited ability to inject anti-inflammatory drugs into the bloodstream, anti-inflammatory drugs are not effective in preventing cytokine release syndrome or or may not be effective in managing cytokine storms. Cytotoxicity and immunosuppression are important factors in the safe use of certain treatments, including T cell therapy and other immunotherapies. Novel strategies for managing cytokine release syndrome and, in particular, cytokine storm are needed. It is considered essential. Summary of the Invention

[0007] In certain embodiments, the present disclosure relates to inhibiting or preventing toxicity of a treatment in an individual. The present invention relates to methods and compositions for treating and preventing toxicity of any of the treatments. In some cases, the methods relate to inhibiting the toxicity of immunotherapy. and a monoclonal antibody or a functional fragment thereof. In some embodiments, the method may be used as part (or all) of an immunotherapy. Any type of therapeutic toxicity can be treated or prevented by the methods of the present disclosure. and compositions. Their toxicity can be at least partially ameliorated by immunotherapy, radiation therapy, and It may be radiation, drug toxicity, oxygen therapy, endocrine therapy, gene therapy, and the like.

[0008] The present disclosure provides, in certain embodiments, a method for treating, for example, systemic inflammatory response syndrome, cytokines, Cytokine release syndromes such as storm, cytokine cascade, or hypercytokinemia This includes the treatment or prophylaxis of a disease or any form of the condition.

[0009] Embodiments of the present disclosure are directed to treating, for example, systemic inflammatory response syndrome, cytokine storm, cytokine Cytokine release syndrome, such as in-cascade or hypercytokinemia, or any of its The present invention relates to a method for determining cytokine levels, including deleterious levels, in an individual, including an individual having a morphology of The harmful level is determined by quantitative measurement of a sample from an individual. The levels can be measured and / or inferred from one or more symptoms. The individual may be screened for symptoms that have directly or indirectly resulted in cytokine release syndrome. Individuals may or may not have been treated. Suffering from an infectious or non-infectious condition that has directly or indirectly resulted in insulin release syndrome They may or may not be affected.

[0010] In a specific embodiment of the present disclosure, the individual is administered an effective amount of fibroblasts and / or fibroblasts. Fibroblast-derived microvesicles are provided for any purpose, and in certain cases, the Fibroblasts are dedifferentiated fibroblasts. Fibroblasts can be dedifferentiated in any manner. However, in specific embodiments, by way of example, one or more histone deacetylase inhibitors Exposure to the agent.

[0011] An embodiment of the present disclosure provides an effective amount of fibroblasts, including dedifferentiated fibroblasts, and / or By providing fibroblast-derived microvesicles to an individual, Methods for decreasing one or more inflammatory cytokines (eg, TNF, etc.) are included.

[0012] In a specific embodiment, the methods of the present disclosure include an effective amount of dedifferentiated fibroblasts. By providing fibroblasts and / or fibroblast-derived microvesicles to an individual, This reduces or eliminates cachexia in the individual. Cachexia in an individual can be caused, for example, by cancer, chemotherapy, or other conditions. It can be caused by any number of reasons, including chronic renal failure, HIV and multiple sclerosis.

[0013] Certain embodiments include an effective amount of fibroblasts, including dedifferentiated fibroblasts, and / or By administering fibroblast-derived microvesicles to an individual, it is possible to The treatment can include, for example, enhancing the response of a recipient individual to one or more treatments. These drugs are prone to toxicity in individuals, such as toxicity associated with excessive cytokine production in It may be any type of treatment, including a medical treatment.

[0014]

[0013] Embodiments of the present disclosure include a method for reducing toxicity of a treatment to an individual, the method comprising: and administering an effective amount of fibroblasts and / or fibroblast-derived microvesicles in combination with the treatment. and / or before and / or after administering the treatment to the individual. The treatment includes immunotherapy, radiation, drug toxicity, oxygen therapy, endoscopy, and the like. The treatment may be one or more of the following: cancer, infectious disease, or gene therapy. and / or treatment for autoimmunity. When the treatment is immunotherapy, the immune The therapeutic may include an antibody or a functional fragment thereof, such as a monoclonal antibody. Any antibody of the present invention may be used. Functional antibody fragments may include, by way of example, scFv. In some cases, treatment (of any kind, including immunotherapy) may be Immunotherapy cells may include stem cells, T cells, NK cells, NKT cells, macrophages, and the like. lymphocytes, B cells, lymphokine-activated cells, tumor-infiltrating lymphocytes, and mixtures thereof. Such cells can express synthetic and / or foreign proteins (e.g., recombinant proteins). The cells may be engineered to express a specific receptor, cytokine, or both. In a specific embodiment, the immunotherapy comprises one or more engineered T A tumor necrosis factor receptor (TCR) or one or more chimeric antigen receptors (CARs) or The TCR or CAR may be one, two, three or more If a cell expresses more than one CAR, the CAR may target two cancer antigens. It may or may not contain the above costimulatory domains.

[0015] In a specific embodiment, fibroblasts are used in the methods and compositions of the present disclosure. The method of the present disclosure further comprises a step of dedifferentiating fibroblasts. In a specific embodiment, the fibroblasts are exposed to a sufficient amount of one or more dedifferentiation agents. When exposed to a dedifferentiating agent, the cells are dedifferentiated or dedifferentiated. In one embodiment, one or more histone deacetylase (HDAC) inhibitors (e.g., valproate) ), exposure to one or more DNMT inhibitors, hypoxia, and / or stem cells or fractions thereof. When valproic acid is used, the valproic acid may be in the range of 1 to 100 micrograms. The fibroblasts may be exposed to the solution at a concentration of 1000 mg / ml for 1 to 72 hours. In some embodiments, fibroblasts are derived from tissues that contain regenerative properties. Examples of tissues include umbilical cord, placenta or a mixture thereof.

[0016] In a specific embodiment, any type of fibroblast used is capable of expressing CD105, CD 117 and / or CD34. In addition, or alternatively, fibroblasts contain expression of rhodamine 123 efflux activity. Such markers and activities may be present in fibroblasts before and / or after dedifferentiation.

[0017] When using microvesicles, the microvesicles are used to The microvesicles may comprise an anion, an exosome-like particle, or a mixture thereof. Ion-exchange chromatography, high performance liquid chromatography (HPLC), or both In a specific embodiment, the method may be used to generate the fibroblasts from a culture of dedifferentiated fibroblasts. Microvesicles are a) CD63; b) CD9; c) MHC I; d) CD56; and and e) express one or more markers selected from the group consisting of: In a specific embodiment, fibroblasts and / or fibroblast-derived microvesicles The antibody has been modified to reduce activation of macrophages. Microvesicles derived from fibroblasts or fibroblasts were incorporated into polymer-reinforced liposomes. obtain.

[0018] In any embodiment, the individual may be provided with an effective amount of activated protein C.

[0019] Embodiments of the present disclosure include determining whether cytokine release in an individual in need of or having been treated is associated with The present invention includes a method of treating or preventing atherosclerosis, the method comprising administering to the patient an effective amount of fibroblasts and / or or fibroblast-derived microvesicles in combination with and / or prior to the treatment. The method includes providing the subject with a cytokine, a cytokine agonist, and / or a cytokine agonist after administering treatment to the subject. The insulin release syndrome may be due to, by way of example, medical treatment, infectious disease or non-infectious disease. In rare cases, non-infectious diseases include graft-versus-host disease (GVHD), acute respiratory distress syndrome (ARDS), and RDS), sepsis, septicemia, pancreatitis, burns, trauma, or hemophagocytic lymphohistiocytosis. Infectious diseases include Ebola, influenza (e.g., avian influenza), and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In the case of influenza, it may be type A, type B or type C. may be influenza C. If cytokine release syndrome is treatment-induced In some cases, the treatment may be an immunotherapy, such as an antibody or a functional fragment thereof. In embodiments, the cytokine release syndrome is systemic inflammatory response syndrome, cytokine release syndrome, The individual may be further defined as having is associated with one or more corticosteroids, one or more biologic therapies, and / or one or more Anti-inflammatory agents may further be provided. In a specific embodiment, the biological therapy is one or more which may include one or more anti-IL6 antibodies.

[0020] Embodiments of the present disclosure include methods for reducing cytokine levels of one or more cytokines in an individual. The method includes a method of reducing the levels of fibroblasts and / or fibroblast-derived fibroblasts, comprising administering an effective amount of fibroblasts and / or fibroblast-derived fibroblasts to the subject. providing the microvesicles to an individual in need of reduction in one or more cytokines. The individual may be suffering from systemic inflammatory response syndrome, cytokine storm, cytokine cascade, may have cytokine release syndrome, which may be further defined as hypercytokinemia or hypercytokinemia. .

[0021]

[0013] Embodiments of the present disclosure include a method of treating cachexia in an individual, the method comprising administering to the individual an effective amount of providing fibroblasts and / or fibroblast-derived microvesicles to the individual. Includes the process.

[0022]

[0013] Embodiments of the present disclosure include a method of increasing the efficacy of a treatment in an individual, the method comprising administering to the individual A body-effective amount of fibroblasts and / or fibroblast-derived microvesicles is administered to the individual. The method includes the step of providing.

[0023] The foregoing has been a summary of the features and techniques of the present disclosure in order to better understand the detailed description that follows. The technical advantages have been outlined quite broadly. Additional features and advantages which form the subject of the claims herein. The disclosed concepts and specific embodiments are within the scope of the present invention. It can be readily used as a basis for modifying or designing other structures to achieve the intended purpose. Those skilled in the art will recognize that such equivalent constructions are set forth in the appended claims. It will be understood by those skilled in the art that such disclosure does not depart from the spirit and scope of the present disclosure. The concepts disclosed herein, both as to organization and method of operation, together with objects and advantages, are The novel features believed to be distinctive will become apparent from the following description when considered in conjunction with the accompanying drawings. Each of these drawings, however, is for purposes of illustration and description only and will be better understood from the following drawings. It is expressly understood that these are provided for illustrative purposes only and are not intended as a definition of the limits of this disclosure. It should be understood.

[0024] In order to provide a more complete understanding of this disclosure, the following description, taken in conjunction with the accompanying drawings, in which: Mention Ming. [Brief description of the drawings]

[0025] [Figure 1] Figure 1 shows the reduction of systemic TNF-alpha by valproic acid treated fibroblasts following PD-1 antibody. Balb / c mice were administered saline (control), antibody to PD-1 (PD-1), or antibody to PD-1 along with Balb / c fibroblasts (100,000 cells intraperitoneally) cultured for 24 hours in 5 micrograms / milliliter valproic acid. Cells and antibodies were injected every other day. Serum TNF-alpha was measured by ELISA.

[0026] [Diagram 2] Figure 2 shows the reduction in lymphokine-activated cell (LAK) lethality by administration of fibroblasts. The left bar is the control, the second bar from the left is LAK, the second bar from the right is LAK+MSC, and the right bar is LAK+fibroblasts. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0027] I. Definition Examples

[0028] In accordance with longstanding patent law practice, the words "a" and "an" are used in conjunction with the term "including." As used herein (including the claims), the term "one or more" refers to the term "one or more" as used herein. Some embodiments include one or more elements, method steps and / or methods of the present disclosure. Any method or composition described herein may consist of or consist essentially of The method may be performed in accordance with any other method or composition described herein. It is contemplated that different embodiments may be combined. Unless the context requires otherwise, the words "comprise" and "contain" "Comprising" and "comprising" refer to the steps or processes described. or elements or steps or elements, but may also include any other steps or is understood to imply no exclusion of any element or step or group of elements. "Consists of" means "including, and including", regardless of what precedes the phrase "consists of." Thus, the phrase "consisting of" means "limited to" the enumerated elements. elements are essential or mandatory and other elements may not be present. "Consisting essentially of" means that any of the elements listed preceding this phrase, and without interfering with the activity or function set forth in this disclosure for the recited elements. "Contributing to" means including elements that are limited to other elements that do not contribute to the invention or to the subject matter. Thus, the phrase "consisting essentially of" means that the recited elements are essential or required. However, other elements are not optional and depend on the activity or May or must not be present depending on whether it affects the function or action of the substance. Throughout this specification, the terms "one embodiment," "an embodiment," and "specific embodiment" are used. "embodiment," "related embodiment," "certain embodiment," "additional embodiment," or " Any reference to a "further embodiment" or combination thereof in relation to that embodiment shall A particular feature, structure, or characteristic described is included in at least one embodiment of the present invention. Thus, the preceding phrases in various places throughout this specification The appearances are not necessarily all referring to the same embodiment. Certain features, structures, or characteristics may be combined in any suitable manner in one or more embodiments. It can be done.

[0029] As used herein, the term "allogeneic" refers to cells of the same species that are genetically distinct from the cells of the host. This refers to cells of the .

[0030] As used herein, the term "autologous" refers to cells derived from the same individual. The term "engraft" as used herein refers to the process by which stem cells join with existing cells of a target tissue. This refers to the process by which a cell is integrated into tissue in vivo via contact.

[0031] As used herein, the terms "about" or "approximately" refer to a quantity. ity, level, value, number, frequency, percentage, dimension, size, amount ), 30, 25, 20, 25, 10, 9, 8, 7, 6, 5, 4, for weight or length 3. Quantity, level, value, number, frequency, or performance that varies by as much as 2 or 1% -Percentage, dimension, size, amount, weight or length. In some embodiments, the terms "about" or "approximately" when used before a numerical value refer to the place of that value. or minus 15%, 10%, 5% or 1%. With respect to processes, the term refers to a range within an order of magnitude, preferably within 5-fold, and more preferably within 10-fold, of a given value. Unless otherwise stated, the term "about" refers to an acceptable range for a particular value. This means that the error is within the allowable error range.

[0032] As used herein, the term "activated fibroblast" refers to a cell that is Changes in metabolic factors, immunological factors, secretion of growth factors, expression of surface markers, and / or or fibers treated with one or more stimuli capable of inducing the production of microvesicles. This refers to blast cells.

[0033] As used herein, the term "activated immune cell" refers to a cell that has one or more Changes in metabolic factors, immunological factors, secretion of growth factors, expression of surface markers, and myelin Immune cells that have been treated with one or more stimuli capable of inducing the production of clonal vesicles Refers to...

[0034] The term "administered" or "administering" as used herein means that a composition is administered to Any method of providing the composition to an individual so as to have the intended effect on the patient. For example, one method of administration is through a medical device (e.g., a catheter, application Indirect mechanisms using, but not limited to, a needle gun, syringe, etc. A second exemplary method of administration is by local tissue administration, oral ingestion, transdermal patch, topical application, or inhalation. These methods involve direct mechanisms such as oral administration or suppositories.

[0035] A "cell culture" may be quiescent, senescent, or (actively) dividing. In cell culture, cells are grown in an artificial in vitro system containing living cells at a suitable temperature. At a temperature of 37° C., typically 37° C., and in an atmosphere typically containing oxygen and CO2 Culture conditions can vary widely from cell type to cell, but the most appropriate culture conditions for a particular cell type are: Different conditions for incubation can result in different phenotypes. The variable is the growth medium, which contains nutrients, growth factor concentrations and other components. The growth factors used to supplement the medium are derived from animal blood. The medium is often, for example, calf serum. The medium may be changed periodically.

[0036] As used herein, the term "dedifferentiated" refers to a cell that has undergone high differentiation after exposure to certain conditions. Refers to cells that have markers of pluripotency and plasticity. For example, induced pluripotent cells are , a dedifferentiated form of fibroblast.

[0037] As used herein, the term "isolated" refers to a substance that is outside the body (e.g., the body (isolated from or derived from a biological sample from the body), stem cells in a naturally occurring state or daughter stem cell population. Biological samples include synovial fluid, blood (e.g., peripheral The tissue may include blood.

[0038] As used herein, the term "microvesicle" refers to a membrane-enclosed intracellular particle. Microvesicles include exosomes, apoptotic bodies, and These include cell parts that have been shed from the cell but remain encapsulated in a membrane.

[0039] The terms "pharmacologically" and "pharmacologically acceptable" as used herein mean , or cause side effects, allergic reactions or other untoward reactions when administered to animals or humans. The term refers to molecular entities and compositions that are not unique to the individual.

[0040] The term "pharmaceutically acceptable carrier" as used herein includes water, ethanol, and the like. polyols (e.g., glycerol, propylene glycol and liquid polyethylene glycols, etc.), suitable mixtures thereof, as well as vegetable oils, coatings, isotonicity agents and and absorption delaying agents, liposomes, commercially available detergents, etc. The supplemental biologically active ingredient may also be used in such a carrier. It can be incorporated into the

[0041] By "population of cells" is meant a collection of at least 10 cells. The population may comprise at least 20 cells, more preferably at least 100 cells, and most preferably at least 100 cells. More preferably, it consists of at least 1000, or even 1 million cells. Because Ming stem cells exhibit the ability to self-renew, they can be expanded in culture to produce billions of cells. A population of can be generated.

[0042] As used herein, the term "purified" as in "purified cells" refers to the presence of other cell types isolated from the subject's body but obtained from that subject's body. "Substantially purified" refers to cells that are still intact, such that the desired cells are at least At least 20%, more preferably at least 50%, and even more preferably at least 7 5%, and most preferably at least 90%, or even 95% concentrated. It means.

[0043] The terms "reduce," "inhibit," "reduce," "suppress," "reduce," "Prevent" and grammatical equivalents (including "lower," "smaller," etc.) are used to describe the procedure. The treated subjects were compared with the untreated subjects with respect to the expression of any symptoms. The amount and / or magnitude of symptoms in the subjects is more pronounced in the medically trained than in untreated subjects. by any amount that is recognized as clinically relevant by any person In one embodiment, the amount of symptoms and / or or the magnitude is at least 10% greater than the amount and / or magnitude of symptoms in untreated subjects. Less / smaller, at least 25% less / smaller, at least 50% less / smaller, At least 75% less / smaller and / or at least 90% less / smaller.

[0044] The term "subject" or "individual" as used herein refers to a person admitted to a medical facility. may be institutionalized or uninstitutionalized and may be treated as an outpatient in a medical facility. An individual may be a person, human or animal. An individual may purchase one or more medical compositions via the internet. An individual may include a human or non-human animal of any age. In the present specification, an individual includes adults and juveniles (i.e., children) and infants. No indication of need for medical treatment is intended, and individuals should not assume any responsibility for clinical or basic scientific research. They may be voluntarily or involuntarily part of an experiment, whether in support of research or for the purposes of that research. A "subject" or "individual" refers to any biological or animal subject that is the subject of a method or material. These include mammals, e.g., humans, experimental animals (e.g., primates, rats, mice, rabbits), livestock (e.g., cattle, sheep, goats, pigs, turkeys) and chickens), household pets (e.g., dogs, cats, and rodents), horses and Transgenic non-human animals are also included.

[0045] "Therapeutic Agents" Have a "Therapeutic Effect" in Modulating Angiogenesis and / or Wound Healing and the amount of therapeutic agent is such that administration of the amount of therapeutic agent is required to modulate angiogenesis. When administered to a subject (e.g., an animal model or a human patient) that has If it is sufficient to cause significant modulation (i.e., increase or decrease), it is considered to be an "angiogenesis It is said to be a "control amount."

[0046] As used herein, the term "therapeutically effective amount" means "an effective amount," "a therapeutically effective "dosage" and / or "effective dose" and are used interchangeably with the term ... It refers to the amount of a compound that elicits the desired biological, cosmetic or clinical response.

[0047] As used herein, the term "toxicity" refers to a pathological change in health, e.g., the physiology of an organ. It refers to a decrease in metabolic activity or damage to cells or tissues.

[0048] II. EXAMPLES OF METHOD EMBODIMENTS OF THE DISCLOSURE

[0049] The present disclosure relates to methods and compositions relating to making therapeutics safe for use in mammals. In certain cases, the methods and compositions relate at least in part to In some cases, this may result in adverse effects in the patient. The treatment includes inhibiting the toxicity of immunotherapy using a novel cellular approach to treat tumors. It may include cancer immunotherapy, such as immunotherapy.

[0050] In one embodiment, a sufficient amount of fibroblasts and / or fibroblast-derived microvesicles is providing a drug to an individual who has toxicity or is susceptible to having or acquiring toxicity; thereby inhibiting or reducing the incidence of toxicity from the treatment in an individual. Disclosed herein is a method for

[0051] Methods, procedures and compositions useful for reducing toxicity associated with immunotherapy, including anti-cancer immunotherapy For example, those that do not substantially impair the therapeutic effect of immunotherapy are disclosed. In the present invention, the individual receiving immunotherapy may be administered one or more immunotherapy treatments prior to and / or after one or more immunotherapy treatments. or simultaneously with and / or following the treatment, fibroblasts and / or fibroblast-derived In some embodiments, the fibroblasts and / or Microvesicles derived from fibroblasts or fibroblasts are capable of plasticity in response to the cytokine environment. These are generated from allogeneic cells with immature phenotypes. Plasticity is a key factor in the activity of tumor-specific immunotherapy. The therapeutic effects of the present invention may include the ability to inhibit systemic antigen-nonspecific inflammatory activity while retaining the therapeutic efficacy of the present invention. In embodiments, the tumor-specific immunotherapeutic activity that is preserved includes, for example, T cells, NKT cells, and the like. These include inflammatory cell and / or NK cell activity.

[0052] In certain embodiments, the toxicity of one or more immunotherapies is determined by fibroblast and / or fibroblast proliferation and / or proliferation. The use of fibroblast-derived microvesicles results in total or partial inhibition. In some cases, the intensity of the toxicity may be reduced. long enough to provide the individual with a sufficient amount of one or more other treatments for sexual or other reasons The above method may be used prophylactically in an individual, or may be used in combination with other methods to slow the onset of toxicity. In a specific embodiment, an individual in need of one or more immunotherapies may include: A sufficient amount of fibroblasts and / or fibroblast-derived microvesicles may be provided. Prior to, during, and / or after, the individual may receive one or more immunotherapies. is provided.

[0053] Fibroblasts or fibroblast-derived microvesicles contain toxicity for immunotherapy It may be provided to an individual for any type of therapeutic toxicity. The toxicity of the method is cytokine release syndrome, and in some cases, individuals develop Cytokine storm (sometimes called cytokine cascade or hypercytokinemia) have or are prone to have severe cases of cytokine release syndrome, including Individuals with insulin release syndrome may have low blood pressure and / or fever. In some cases, cytokine release syndrome is characterized by increased production of one or more cytokines. In a specific embodiment, the cytokines are: a) TNF-alpha; b) IL-1 Beta;c)IL-6;d)IL-33;e)CRP;f)IL-17;g)IL-2; h) IL12; i) IL-18; j) HMGB-1; k) interferon gamma; l) interferon alpha; and m) combinations thereof. .

[0054] In certain embodiments, the individual is diagnosed with a cancer that has been induced by administration of one or more cancer immunotherapeutic agents. have or are predisposed to cytokine release syndrome, which is a common cause of cancer immunotherapy The administration may or may not be new to the individual. In one embodiment, the cancer immunotherapy comprises a chimeric antigen receptor (CAR)-specific immune cell. Such CAR-specific immune cells can be CAR-specific NK, NKT or T cells. The CAR can be directed to any antigen, including any cancer antigen, such as any tumor antigen. AR is 1st generation type, 2nd generation type, 3rd generation type or later In a specific embodiment, the generation type may be C ARs are composed of one, two or more co-stimulatory receptors, including at least CD28, 4-1BB, etc. Includes intense domains.

[0055] In some embodiments, the individual is diagnosed with sarcoma caused by one or more infectious agents. have or are susceptible to influenza A release syndrome. b) Avian influenza; c) Severe acute respiratory syndrome (SARS); d) Epstein-Barr virus Barr virus-associated hemophagocytic lymphohistiocytosis (HLH); e) bacterial sepsis; f) Gram Negative sepsis; g) Dengue virus; h) Malaria; i) Ebola virus; j) Smallpox virus k) systemic gram-negative bacterial infections; and l) combinations thereof. It may or may not be so.

[0056] In certain embodiments, the individual has one or more non-infectious causes. Having or prone to having cytokine release syndrome. Non-limiting examples include: a) blood cell Phagocytic lymphohistiocytosis (HLH); b) Macrophage activation syndrome, which is a sporadic form of HLH (MAS), c) chronic arthritis; d) systemic juvenile idiopathic arthritis (sJIA); e) still disease; f) cryopyrin-associated periodic syndrome (CAPS); g) familial cold autoinflammatory syndrome (FCAS); h) familial cold urticaria (FCU); i) Muckle-Wells syndrome (MW S);j) Chronic Infantile Neuropathic Cutaneous and Articular Disease (CINCA) syndrome;k) Inherited in the NLRP3 gene Cryopyrinopathies including multiple or novel gain-of-function mutations hy);l) inherited autoinflammatory disorders;m) acute pancreatitis;n) severe burn injury;o) acute radiation p) trauma; q) acute respiratory distress syndrome; r) systemic inflammatory response syndrome; and s) and combinations thereof.

[0057] In certain embodiments, the individual is diagnosed with cytokine release syndrome or cytokine stomatitis. The individual may have or develop one or more immunomodulatory cancer therapies, e.g., CA Treatments containing immune cells expressing R (including at least NK, NKT or T cells) May or may not have received (and / or have received and / or In certain embodiments, the method includes the step of: The method includes administering one or more compositions comprising fibroblast-derived microvesicles, Administration of the compound inhibits the development of cytokine release syndrome or cytokine storm in an individual. or to impair, or to reduce the incidence of, or to delay the occurrence of, or Prevents the development of fibroblasts and / or fibroblast-derived microvesicles. The compositions can be administered prior to, concurrently with, or after an immunomodulatory therapy, such as a CAR T cell therapy. In some embodiments of the present disclosure, the method may include administering a fibroblast cell therapy to a subject, the subject being provided with a fibroblast cell therapy, and / or the subject being provided with a fibroblast cell therapy. Microvesicles derived from cells and / or fibroblasts are described in U.S. Patent Application Publication No. US2002 / 0133663. / 0086027, which is incorporated herein by reference in its entirety. Reduces cytokine release signals, including those that enhance T cell memory responses. It is administered at the same time as immunotherapy.

[0058] Treatment with fibroblasts and / or fibroblast-derived microvesicles enhances cytokine production. Treatment-related symptoms, including cytokine storm, a severe symptom of insulin release syndrome, It may be used within the context of the present disclosure to treat any type of toxicity. Cytokine storms are also a concern following infectious or non-infectious stimuli. Numerous proinflammatory cytokines (e.g., interleukin-1 (IL-1), IL- 6. IL-8, gamma interferon (gamma IFN), macrophage inflammatory protein -1α (MIP-1α), tumor necrosis factor-alpha (TNFα), or a combination thereof The result is hypotension, hemorrhage, and ultimately multiple organ failure. H1N1 influenza pandemic and the more recent avian influenza H5N1 infections The relatively high mortality rate in young people with presumably healthy immune systems in This syndrome is caused by a storm of infection. Barr virus-associated hemophagocytic lymphohistiocytosis, gram-negative sepsis, malaria, and many other It is known to occur in advanced or terminal cases of many other infectious diseases, including Ebola infection. Cytokine storms can occur in patients with acute pancreatitis, severe burns or trauma, or acute respiratory distress. It can also result from non-infectious causes such as cytokine release syndrome, in particular Novel strategies for managing the cytokine storm are needed.

[0059] In one embodiment of the present disclosure, fibroblasts and / or fibroblast-derived microbes are Cyclin plays an important role in the immunological cascade associated with cytokine release syndrome. It is used to reduce the activation of macrophages, which play a key role in the development of In embodiments, the present disclosure provides fibroblasts and / or fibroblast-derived microvesicles. The present study proposes selective targeting of the antibody to macrophages and / or other cells of the reticuloendothelial system. In one embodiment, the fibroblasts are provided with the ability to deliver fibroblasts to the reticuloendothelial system. In one embodiment, the drug is administered in the form of a highly potent liposomal preparation. Uses of polymer-reinforced liposomes are provided. Examples of polymers useful for the preparation of glycerol include poly-l-lysine, polyamidoamine dendrimers, and and polyethereneimines. Descriptions of polymer-reinforced liposomes can be found in the literature. It is described in.

[0060] In certain embodiments, the present disclosure relates to fibroblasts and / or fibroblast-derived myeloid leukocytes. Any symptom including at least systemic inflammatory response syndrome (SIRS) caused by administration of clovesicles The present invention provides for the prevention or amelioration of at least one symptom of a form of cytokine release syndrome. In the context of the disclosure, SIRS may be caused by infectious, traumatic or other causes. is a term that characterizes an inflammatory syndrome caused by inflammation in which the patient meets the following criteria: At least two of the following: 1) body temperature less than 36°C or more than 38°C; 2) pulse rate greater than 90 beats / min 3) Tachypnea with respiratory rate of more than 20 breaths per minute or 4.3 kPa (32 m 4) Arterial partial pressure of carbon dioxide less than 4000 cells / mm 3 (4×10 9 cells / L) Less than or equal to 12,000 cells / mm 3 (12×10 9 White blood cell count > 100 cells / L; or Presence of >10% immature neutrophils (band-shaped). In sepsis, active infection is present, and SIR S is distinct from sepsis. These patients may develop acute renal failure; pulmonary failure; shock and / or The term "septic shock" refers to the inability of a patient to receive adequate fluid resuscitation. Systolic blood pressure <90mmHg despite receiving blood pressure medication and vasopressors / inotropes It refers to the state of having high blood pressure.

[0061] The main events that can occur with cytokine release syndrome are progression to SIRS and The main events in the progression to subsequent multiple organ failure, etc., within the context of this disclosure, are: It is inhibited by fibroblasts and / or fibroblast-derived microvesicles. Such events may, in certain cases, include: a) systemic activation of the inflammatory response; b) endothelial activation and proliferation, associated with consumption of anticoagulants and fibrinolytic factors and initiation of the coagulation cascade; c) activation of complement; and d) organ failure and death. These pathological events appear to be related to each other; for example, complement activation has procoagulant properties. In cancer patients, SIRS is triggered by several factors. Many patients receive immunosuppressive chemotherapy / radiation therapy that promotes opportunistic infections. Furthermore, given that approximately 40-70% of patients are cachectic, This low-grade inflammation triggers further bacterial / injury-induced inflammatory cascades. Finally, tumors, both by themselves and through interactions with host factors, may enhance S. Systemic effects such as IL-1, IL-6, and TNF-alpha that may predispose to IRS Without wishing to be bound by theory, it has been demonstrated that the inflammatory mediators However, one of the mechanisms of action of fibroblasts is to suppress cachexia and improve response to treatment. To increase the likelihood, it may be a reduction in inflammatory cytokines such as TNF.

[0062] In one embodiment, the fibroblasts are administered X-linked fibroblast inhibitors to inhibit cytokine release syndrome. It is used in combination with igris (activated protein C (APC)). Xigris is angio Stimulation of barrier function via the poietin / Tie-2 axis mediates protection from apoptosis by activating endothelial cell protective mechanisms mediated by endothelial cell adhesion and by reducing local coagulation. The basis for Xigris' approval has been questioned by some. Additionally, it is considered to be a major concern in oncology-related sepsis, particularly in white blood cells where bleeding is a major concern. In fact, it is often counter-indicated in In the Phase III trial of Xigris, hematopoietic transplant patients were excluded.

[0063] One of the main causes of death associated with SIRS in its most advanced stages is disseminated intravascular coagulation ( DIC is a microcirculatory dysfunction that is associated with SIRS. Whether it is endotoxin, TLR agonists or damage-related agonists such as HMGB-1, All of these signals can activate the endothelium systemically. In this case, the endothelial response to such mediators is localized and serves to contain the infection. provides a useful mechanism for preventing and enabling immune attack. The fact that the response is systemic can have dire consequences, including organ failure. Characteristics of the response include: a) upregulation of tissue factor (TF), leading to a procoagulant state; and endothelial coagulation inhibitors such as protein C and the antithrombin system. b) high expression of adhesion molecules that induce neutrophil extravasation; c) low fibrinolytic capacity and d) increased vascular permeability / unresponsiveness to vasodilators and vasoconstrictors. A review of molecular signals related to SIRS-induced endothelial dysfunction has been published. One of the important factors involved is NF-kB. Nuclear translocation of NF-kB is a key factor in the formation of thrombus. Associated with endothelial upregulation of pro-fibrinolytic molecules and inhibition of fibrinolysis. In a recent study, Song et al. administered exogenous i-kappa B (an NF-kB inhibitor) to vascular By generating transgenic mice that express specific transgenic genes in the These transgenic mice were then cultured in a 3-well platelet-free, 5-HT- treated control group, and the NF-kB expression was selectively inhibited in the endothelium. In contrast to wild-type animals, endothelial cells from mice were significantly more susceptible to endothelial protein upregulation after endotoxin challenge. Substantially suppresses tissue factor expression while preserving expression of thrombomodulin and phosphokinin C receptor. Furthermore, expression of NF-B was reduced by TNF-alpha as a result of TACE activity. associated with the production of

[0064] immunotherapy

[0065] In some embodiments, the methods and compositions encompassed herein provide for immunotherapy. The toxicity of the method is addressed. This toxicity is caused by the following factors: stem cells, T cells, NK cells, NKT cells, macrophages, and Phages, B cells, lymphokine-activated cells, tumor-infiltrating lymphocytes and mixtures thereof Such cells may be relevant for any kind of cell therapy, including the delivery of synthetic proteins and and / or exogenous proteins (e.g., receptors, cytokines, or both) ), or may not be engineered to express the In some embodiments, any immunotherapy, including cell therapy, may be performed using engineered antigen receptors (e.g., The present invention may include cells having a specific receptor, such as a CAR, a TCR, a chimeric cytokine receptor, etc.

[0066] In the art, CAR-T cell administration with pentoxifylline or various formulations is Treatment of toxicity associated with CAR-expressing cells may be applied to a variety of CAR-expressing cells. In one embodiment, In this way, CAR-expressing cells can bind to a specific antigen via an antibody or antibody fragment that targets that antigen. In another embodiment, the antibody comprises a monoclonal antibody that binds to an epitope of an antigen. In another embodiment, the antibody is a polyclonal antibody. In some embodiments, the antibody fragment is a single chain variable fragment (scFv).

[0067] In certain embodiments, the CAR-expressing cell of the composition as disclosed herein comprises: Bind to one or more tumor-associated antigens (TAA). Cells can be delivered with multiple CAR molecules or a single The CAR molecule may target one antigen or may target two or more antigens. The antigen may be targeted.

[0068] Examples of CAR-expressing cells herein can be exemplified as T cells, but include at least Any immune cell, including NK cells or NKT cells, can be modified with one or more CARs. In one embodiment, the tumor-associated antigen is cell surface-associated mucin 1 (MUC1) or Polymorphic epithelial mucin (PEM), variant early intratumoral arginine-rich (Armet), and fever syndrome Heat shock protein 60 (HSP60), calnexin (CANX), methylenetetrahydrofolate Folate dehydrogenase (NADP+-dependent) 2, methenyltetrahydrofolate cyclohydrolase fibroblast activation protein (FAP), matrix metallopeptide MMP6, B-melanoma antigen-1 (BAGE-1), N-acetylglucosamine Aberrant transcripts of GnTV (Gnyltransferase V), Q5H943, and carcinoembryonic antigen ( CEA), Pmel, kallikrein-4, mammaglobin-1, MART-1, GPR1 43-OA1, prostate-specific antigen (PSA), TRP1, tyrosinase, FGP-5, NE U proto-oncogene, Aft, MMP-2, prostate-specific membrane antigen (PSMA), telomerase-related Protein-2, prostatic acid phosphatase (PAP), uroplakin II or protein Other types of CAR-T cells are known, including those that target leukemia. If you want to destroy B cells, such as those in In another embodiment, the CAR is a T cell that binds to RO In another embodiment, the CAR binds to NY-ESR1, CD22, or GD2. In another embodiment, the CAR binds to a MAGE family protein. In another embodiment, the CAR binds to mesothelin. In another embodiment, the CAR binds to c-erbB2. It binds to tumor-specific mutant antigens such as FV600E mutations and BCR-ABL translocations. In another embodiment, the CAR is directed against EBV in HD, HPV in cervical cancer, and Binds to tumor-specific viral antigens, such as polyomavirus in Merkel carcinoma. In one embodiment, the CAR T cells bind to Her2 / neu. In this study, the CAR T cells bind to EGFRvIII.

[0069] In one embodiment, the chimeric antigen receptor (CAR) T cells are directed against the CD19 antigen. In another embodiment, the CAR binds to the CD22 antigen. In another embodiment, the CAR binds to an alpha folate receptor. In another embodiment, the CAR binds to CAIX. In another embodiment, the CAR binds to CD20. In one embodiment, the CAR binds to CD23. In another embodiment, the CAR binds to In another embodiment, the CAR binds to CD24. In another embodiment, the CAR binds to CD30. In one embodiment, the CAR binds to CD33. 38. In another embodiment, the CAR binds to CD44v6. In one embodiment, the CAR binds to CD44v7 / 8. In another embodiment, the CAR binds to CD171. In another embodiment, the CAR binds to carcinoembryonic antigen (CEA). In another embodiment, the CAR binds to EGFRvIII. In another embodiment, the CAR binds to EGP-40. In one embodiment, the CAR binds to EphA2. In another embodiment, the CAR binds to In another embodiment, the CAR binds to Erb-B2, 3, 4. In another embodiment, the CAR binds to Erb-B3 / 4. In another embodiment, the CAR binds to a fetal acetylcholinesterase (FBP). In another embodiment, the CAR binds to the G.sub.D2 receptor. In another embodiment, the CAR binds to G.sub.D3. In another embodiment, the CAR binds to HER2. In another embodiment, the CAR binds to IL-11R alpha. In another embodiment, the CAR binds to IL-13Ralpha1. In one embodiment, CAR binds to KDR. In another embodiment, the CAR binds to Lewis Y. In an embodiment, the CAR binds to the L1-cell adhesion molecule. The CAR binds to MAGE-A1. In another embodiment, the CAR binds to mesothelin. In another embodiment, the CAR binds to a CMV infected cell. In another embodiment, the CAR binds to MUC1. In another embodiment, the CAR binds to an NKG2D ligand. In an embodiment, the CAR binds to NY-ESO-1 (amino acids 157-165). In another embodiment, the CAR binds to carcinoembryonic antigen (h5T4). In another embodiment, the CAR binds to PSMA. In another embodiment, the CAR binds to ROR1. In another embodiment, the CAR binds to TAG-72. In another embodiment, the CAR binds to B In another embodiment, the CAR binds to CA9. In this state, CAR binds the alpha v beta 6 integrin In another embodiment, the CAR binds to 8H9. In another embodiment, the CAR binds to fetal acetylcholine. In another embodiment, the chimeric antigen receptor (CAR) T The cells target the CD19 antigen and are involved in the development of B-cell malignancies, including ALL, follicular lymphoma, CLL, and In another embodiment, the CAR T cells target the CD22 antigen and provide therapeutic benefit to subjects with B cell malignancies. In another embodiment, the CAR T cells have an alpha folate receptor or Targeting the acid receptor alpha to provide therapeutic benefit to subjects with ovarian or epithelial cancer In another embodiment, the CAR T cell has CAIX or G250 / CAI X is targeted and has a therapeutic effect on a subject with renal cell carcinoma. CAR T cells target CD20 and are used to treat lymphomas, B-cell malignancies, and B-cell lymphomas. The present invention is directed to treating subjects with idiopathic bronchitis, mantle cell lymphoma, and indolent B-cell lymphoma. In another embodiment, the CAR T cells target CD23 and inhibit CL In another embodiment, the CAR T cells have a therapeutic effect on a subject having L. The present invention targets CD24 and has a therapeutic effect on subjects with pancreatic adenocarcinoma. In one embodiment, CAR T cells target CD30 and treat lymphoma or Hodgkin's lymphoma. In another embodiment, the CAR T cells have a therapeutic effect on a subject having In another embodiment, the method targets CD33 and has a therapeutic effect on subjects with AML. In this study, CAR T cells target CD38 and are administered to subjects with non-Hodgkin's lymphoma. In another embodiment, the CAR T cells target CD44v6. In another embodiment, the present invention targets the tumor and has a therapeutic effect on subjects with some malignant tumors. In this study, CAR T cells were targeted to CD44v7 / 8 and administered to subjects with cervical cancer. In another embodiment, the CAR T cells target CD123 and have a therapeutic effect. and has a therapeutic effect on a subject with a myeloid malignancy. CAR T cells target CEA and have therapeutic effects in subjects with colorectal cancer. In another embodiment, the CAR T cells target EGFRvII and inhibit glioblastoma. In another embodiment, the CAR T cells have a therapeutic effect on a subject having a tumor. The present invention targets EGP-2 and has therapeutic effects on subjects with multiple malignancies. In one embodiment, the CAR T cells target EGP-40 and are In another embodiment, the CAR T cells have a therapeutic effect on the subject. 2 and has a therapeutic effect on a subject with glioblastoma. In this study, CAR T cells target Erb-B2 or ErbB3 / 4 to treat advanced cancers, including breast cancer. The present invention has a therapeutic effect on subjects with prostate cancer, colon cancer, and various tumors. In this study, CAR T cells target Erb-B 2, 3, and 4 and treat patients with breast cancer, etc. In another embodiment, the CAR T cells target FBP. and has a therapeutic effect on a subject with ovarian cancer. T cells target fetal acetylcholine receptors and respond to subjects with rhabdomyosarcoma In another embodiment, the CAR T cells inhibit G.sub.D2 and have a therapeutic effect. Targeting and Treating Subjects with Neuroblastoma, Melanoma, or Ewing's Sarcoma In another embodiment, the CAR T cells target GD3 and inhibit melanoma. In another embodiment, the CAR T cells have a therapeutic effect on a subject with a rheumatoid arthritis. The present invention targets HER2 and is directed to treating patients with medulloblastoma, pancreatic adenocarcinoma, glioblastoma, osteosarcoma, or ovarian cancer. In another embodiment, the CAR T cells have a therapeutic effect on the subject. It targets AA and has a therapeutic effect on subjects with melanoma. In this study, CAR T cells target IL-11R alpha and are used in subjects with osteosarcoma. In another embodiment, the CAR T cells have a therapeutic effect against IL-13R- Targeting pha-1 and providing therapeutic benefit to subjects with glioma, glioblastoma, or medulloblastoma In another embodiment, the CAR T cells have the IL-13 receptor alpha 2 The present invention targets the IL-11 receptor agonist, IL-11 receptor agonist, and has a therapeutic effect on subjects with some malignant tumors. In this study, CAR T cells target KDR and the neovasculature of tumors. In another embodiment, the method comprises administering to a subject having a tumor a therapeutic effect. AR T cells target kappa-light chains and inhibit B-cell malignancies (B-NHL, CLL). In another embodiment, the CAR T cells have a therapeutic effect on a subject having Targeting ewis Y to treat subjects with various carcinomas or tumors of epithelial origin In another embodiment, the CAR T cells target the L1-cell adhesion molecule. and has a therapeutic effect on a subject with neuroblastoma. AR T cells target MAGE-A1 or HLA-A1 MAGE A1 and melanoma In another embodiment, the CAR T cell has a therapeutic effect on a subject having a glioma. The cells target mesothelin and have a therapeutic effect on subjects with mesothelioma. In embodiments, the CAR T cells target CMV-infected cells and are administered to subjects with CMV. In another embodiment, the CAR T cells target MUC1. and has a therapeutic effect on a subject with breast cancer or ovarian cancer. In this study, CAR T cells targeted MUC16 and demonstrated therapeutic efficacy in subjects with ovarian cancer. In another embodiment, the CAR T cells target an NKG2D ligand, It has therapeutic effects on subjects with myeloma, ovarian and other tumors. In this study, CAR T cells were targeted to NY-ESO-1(157-165) or HLA-A 2 Targeting NY-ESO-1 and having therapeutic effect on subjects with multiple myeloma In another embodiment, the CAR T cells target carcinoembryonic antigen (h5T4) and In another embodiment, the CAR T The cells target PSCA and have a therapeutic effect on subjects with prostate cancer. In embodiments, the CAR T cells target PSMA and the vasculature of prostate cancer / tumors. In another embodiment, the CAR T cells have a therapeutic effect on a subject having Targeting ROR1 to Treat Subjects with B-CLL and Mantle Cell Lymphoma In another embodiment, the CAR T cells target TAG-72 and In another embodiment, the present invention has a therapeutic effect on a subject having adenocarcinoma or gastrointestinal cancer. CAR T cells target VEGF-R2 or other VEGF receptors to induce tumor proliferation and Targeting the living vasculature has a therapeutic effect on tumor-bearing subjects. In another embodiment, the CAR T cells target CA9 and are administered to a subject with renal cell carcinoma. In another embodiment, the CAR T cells target CD171. In another embodiment, the method is directed to treating renal neuroblastoma. In this study, CAR T cells targeted NCAM and were used to treat subjects with neuroblastoma. In another embodiment, the CAR T cells have a therapeutic effect. The present invention relates to a method for treating rhabdomyosarcoma by targeting a rhabdomyosarcoma-associated protein 1 (HMP-1) in a subject having rhabdomyosarcoma. In this study, CAR binds to angiogenic factors and thereby targets the tumor vasculature. In one embodiment, the angiogenic factor is VEGFR2. In another embodiment, the angiogenic factor is endoglin. The angiogenic factors that are involved are angiogenin, angiopoietin-1, Del-1, and fibroblast growth factor receptor 1 (FGR1). Follicular growth factors: acidic (aFGF) and basic (bFGF); follistatin; granulocyte colony knee stimulating factor (G-CSF); hepatocyte growth factor (HGF) / cell scattering factor (scatter); r factor (SF); interleukin-8 (IL-8); leptin; midk In; Placental growth factor; Platelet-derived endothelial cell growth factor (PD-ECGF); Platelet-derived growth factor Cause child-BB (PDGF-BB); pleiotrophin (PTN); progranulin; pro Proliferin; Transforming growth factor-alpha (TG F-alpha); Transforming Growth Factor-beta (TGF-beta); Tumor Necrosis Factor-alpha (TNF-alpha); vascular endothelial growth factor (VEGF) / vascular permeability factor In another embodiment, the angiogenic factor is an angiogenic protein. In one embodiment, the growth factor is an angiogenic protein. In the present invention, angiogenesis inhibitors for use in the compositions and methods disclosed herein are Proteins include fibroblast growth factor (FGF); VEGF; VEGFR and neuropilin. 1 (NRP-1); angiopoietin 1 (Ang1) and Tie2; platelet-derived growth factor PDGF; BB-homodimer and PDGFR; transforming growth factor- beta (TGF-.beta.), endoglin and TGF-beta receptor; monocyte chemotaxis Chemical motility protein-1 (MCP-1); integrin alpha V beta 3, alpha VE-cadherin and CD 31;Ephrin;Plasminogen activator;Plasminogen activator inhibitor-1; Nitric oxide synthase (NOS) and COX-2; AC133; or Id1 / Id3 In one embodiment, the compositions and methods disclosed herein are used The angiogenic proteins that mediate this are the angiopoietins, which In one embodiment, angiopoietin 1, angiopoietin 3, angiopoietin 4 or angiopoietin 6. In one embodiment, endoglin is CD1 05;EDG;HHT1;ORW; or also known as ORW1. In one embodiment In another embodiment, endoglin is a TGF-beta coreceptor. AR T cells bind to antigens associated with an infectious agent. The agent is Mycobacterium tuberculosis. The Mycobacterium tuberculosis-associated antigen is an antibody Hara85B, lipoprotein IpqH, putative ATP-dependent helicase, protein of unknown nature Rv0476 / MTO4941 precursor or unknown protein Rv1334 / MT13 76 precursor. In another embodiment, the CAR is attached to an antibody. In this study, CAR T cells are "antibody-associated T cell receptors" (ACTRs). According to an embodiment, the CAR T cells are universal CAR T cells. In one embodiment, the CAR T cells having an antibody receptor are activated before the antibody is administered. The CAR T cells are then administered either after or at the same time that the antibody is administered. In another embodiment, the antibody is In another embodiment, the antibody is directed against a tumor cell antigen. In another embodiment, the antibody is rituximab.

[0070] In another embodiment, fibroblasts or preparations thereof are administered to a site following administration of a therapeutic antibody. The antibody may be of any type. The antibodies include Fab', Fab, F(ab')2, single domain antibodies (DAB), F Antibodies include fragments such as polyclonal Fv, scFv (single chain Fv), etc. In one embodiment, the antibody is directed to ERBB2. Trastuzumab (Herceptin; Genentech), a humanized IgG1 that In another embodiment, the antibody is a VEGF-directed humanized IgG1. Vacizumab (Avastin; Genentech / Roche). In the study, the antibody was Cetuximab, a chimeric human-mouse IgG1 directed against EGFR. Another implementation is Erbitux (Bristol-Myers Squibb). In one embodiment, the antibody is panitumumab (Vect), a human IgG2 directed against EGFR. In another embodiment, the antibody is directed against CTLA4. ipilimumab (Yervoy; Bristol-Myers Squibb), an IgG1 inhibitor bb). In another embodiment, the antibody is a humanized IgG1 directed against CD52. One alemtuzumab (Campath; Genzyme). , the antibody was ofatumumab (Arzerra), a human IgG1 directed against CD20; In another embodiment, the antibody is a humanized I directed against CD33. The other is gemtuzumab ozogamicin (Mylotarg; Wyeth), which is a type of gG4. In an embodiment, the antibody is Brentuximab, a chimeric IgG1 directed against CD30. Another example is buvedotin (Adcetris; Seattle Genetics). In one embodiment, the antibody is a 90Y-labeled immobilized antibody that is a murine IgG1 directed against CD20. Momab tiuxetan (Zevalin; IDEC Pharmaceuticals) In another embodiment, the antibody is a murine IgG2 directed against CD20. 13 1 I-labeled tositumomab (Bexxar; GlaxoSmithKline). In an embodiment, the antibody is directed against vascular endothelial growth factor receptor-2 (VEGFR-2). In another embodiment, the antibody is ramucirumab (Cyram za injection, Eli Lilly and Company), blinatumomab (BLI NCYTO, Amgen Inc.), pembrolizumab (KEYTRUDA, Merck K Sharp & Dohme Corp.), obinutuzumab (GAZYVA, Gene ntech, Inc.; formerly known as GA101), Pertuzumab Injection ( PERJETA, Genentech, Inc.) or denosumab (Xgeva, Am In another embodiment, the antibody is basiliximab (Simugen Inc.). lect; Novartis). In another embodiment, the antibody is daclizumab ( In another embodiment, the CAR T cell is The antibodies used herein can be any of the tumor or tumor-specific antibodies described herein and / or known in the art. In another embodiment, the CAR T cell is directed to a cancer antigen or a portion thereof. In another embodiment, the antibody that is bound to the cell is directed to a tumor-associated antigen. The antibodies bound to RT cells are directed against tumor-associated antigens, which are angiogenesis factors, or parts of such antigens. It is an elephant.

[0071] Embodiments of the present disclosure relate to fibroblasts and / or fibroblast-derived microvesicles. This includes immunotherapy preparations used together with Organizations that perform and / or use fibroblasts and / or fibroblast-derived microenvironments However, in some cases In immunotherapy, fibroblasts and / or fibroblast-derived microvesicles are prepared. In some cases, immunotherapy is obtained from organizations that do not and / or do not use Based on having one or more symptoms of a medical condition that may be effective, and / or based on the individual being Recognizing an individual as in need of immunotherapy based on having a formal diagnosis of the condition. Immunotherapy and treatment with fibroblasts and / or fibroblast-derived microvesicles , may be prepared and provided to an individual at the same time and / or at different times. Treatment with fibroblasts and / or fibroblast-derived microvesicles is an immunotherapy As another example, immunotherapy may be administered to an individual for a sufficient length of time prior to initiating immunotherapy. The method involves initiating treatment with fibroblasts and / or fibroblast-derived microvesicles. The vaccine may be provided to the individual for a sufficient length of time prior to immunization. The route of delivery of the therapy is via fibroblasts and / or fibroblast-derived microvesicles. This may or may not be the same as the route of delivery of the therapeutic. , immunotherapy and / or fibroblasts and / or fibroblast-derived microvesicles Treatment with the clotting agent may be provided to an individual once, or more than once. If administered, the second or subsequent immunotherapy administration may be of a different type and / or amount of immunotherapy. For example, the immunotherapy may include different antibodies, or in the case of CAR therapy, The CAR may be directed to a different antigen than the first CAR therapy. If treatment with fibroblast-derived microvesicles is provided more than once, the second Subsequent administration of fibroblast and / or fibroblast-derived microvesicle treatments , may contain different fibroblasts and / or fibroblast-derived microvesicles. For example, the markers may include one or more different markers, may be derived from different tissue sources, may be derived from different microorganisms, The antibodies may include vesicles, may be derived from different individuals, combinations thereof, and the like.

[0072] III. Fibroblasts and Their Creation or Manipulation

[0073] In a specific embodiment, the fibroblasts may be pretreated for any purpose prior to delivery to the individual. For example, fibroblasts can be engineered to undergo dedifferentiation. For example, fibroblasts may be pretreated with the above agents for use in the therapeutic methods of the present disclosure. Prior to use, patients should be treated with one or more histone deacetylase inhibitors (e.g., valproic acid); one or more DNA methyltransferase inhibitors; exposure to stem cells; hypoxia; and their combination A particular treatment for fibroblasts may be administered for a particular period of time (e.g. As, 1 to 72 (or 1 to 48 or 1 to 36 or 1 to 24 or 1 to 18 or 1~12 or 1~6 or 6~72 or 6~48 or 6~36 or 6~24 or is 6~18 or 6~12 or 12~72 or 12~48 or 12~36 or 1 2~24 or 12~18 or 18~72 or 18~48 or 18~36 or 1 8~24 or 24~72 or 24~48 or 24~36 or 36~72 or 3 1 to 100 (or for example 1 to 75 or 1 to 50 or 1 to 6) hours) ~25 or 10~100 or 10~75 or 10~50 or 10~25 or 2 5~100 or 50~100 or 75~100 or 25~100 or 25~75 or 25-50 or 50-100 or 50-75 or 75-100) micrograms An exemplary histone deacetylase inhibitor, Valproate, at a concentration of 1000g / milliliter Treatment with rotic acid may be used to induce dedifferentiation.

[0074] In certain embodiments, the fibroblasts are not any tissue that contains fibroblasts. They may be derived from a particular tissue. In a specific embodiment, fibroblasts have regenerative properties. Derived from one or more tissues. In one example, fibroblasts are derived from placental tissue or umbilical cord tissue. In either method, the fibroblasts are freshly cultured prior to manipulation for the disclosed method. It can be extracted.

[0075] In some cases, fibroblasts, whether derived from a particular tissue or not, One or more specific markers may be selected for inclusion. Examples include CD105, Fibroblasts selected for expression of CD117 and / or CD34 are included. In some cases, fibroblasts express CD105, CD117 and / or CD3 Alternatively or in addition to expressing 4, expression of rhodamine 123 efflux activity The selection is made according to the

[0076] In one embodiment, allogeneic fibroblasts are cultured in a non-manipulated manner (e.g., in vitro). administered to an individual without prior exposure to one or more specific agents, such as feron gamma However, they are selected from sources that are naturally characterized by immunomodulatory activity (e.g., placental fibroblasts or In another embodiment of the present disclosure, any fibroblast may be: Culture under conditions that can induce retrodifferentiation to confer an immature phenotype to fibroblasts and their immature phenotype correlates with high anti-inflammatory and / or immunomodulatory potential. For example, fibroblasts may be treated with one or more histone deacetylase inhibitors, such as valproate. The presence of acid (Moon et al., 2008; Huang et al., 2011). In addition to HDAC inhibitors, other inhibitors that induce dedifferentiation of fibroblasts can be used. Means (e.g., 8-Br-cAMP (Wang et al., 2011); M-CSF treatment (Li et al., 2016); exposure to rebelecin (Li et al., 2 016); and / or exposure to stem cell extracts (Xiong et al., 2014 )) may also be used in the context of the present disclosure. Markers (e.g., CXCR4, VEGFR-2, CD34 and / or CD 133) and intracellular markers (e.g., SOX-2, NANOG and / or OC This can be done by evaluation of the following:

[0077] Fibroblasts used in the methods and compositions, or in the production of certain methods and compositions The alveoli are composed of salivary gland mucous cells, salivary gland serous cells, von Ebner's gland cells, mammary gland cells, lacrimal gland cells, Ear canal gland cells, eccrine sweat gland dark cells, eccrine sweat gland clear cells, apocrine sweat gland cells, molar gland cells alveolar sebaceous gland cells, Bowman's gland cells, Brunner's gland cells, seminal vesicle cells, prostate cells, bulbourethral gland cells cells, Bartholin's gland cells, Little gland cells, endometrial cells, isolated goblet cells, mucosal cells of the stomach wall alveolar cells, gastric gland zymogen cells, gastric gland acid secreting cells, pancreatic acinar cells, Paneth cells, type II alveolar cells, Lara cells, growth hormone producing cells, lactogenic hormone producing cells, thyrotropin producing cells cells, gonadotropes, adrenocorticotropes, cells of the intermediate lobe of the pituitary gland, Large cell neurosecretory cells, intestinal cells, airway cells, thyroid epithelial cells, parafollicular cells, parathyroid cells, Chief parathyroid cells, eosinophilic cells, adrenal cells, chromaffin cells, Leydig cells, theca interna Cells, luteal cells, granulosa luteal cells, theca luteal cells, juxtaglomerular cells, macula densa cells, perivascular Pole cells, mesangial cells, vascular endothelial fenestrated cells and lymphatic endothelial fenestrated cells, vascular endothelial connection cells endothelial and lymphatic endothelial continuous cells, endothelial and lymphatic endothelial splenocytes, synovial cells alveoli, serosal cells (lining the peritoneal, pleural and pericardial cavities), squamous cells, columnar cells, dark cells , vestibular membrane cells (the inner wall of the endolymphatic space of the ear), basal cells of the stria vascularis, peripheral cells of the stria vascularis (the endolymphatic space of the ear) Inner lining of the lumen, Claudius cells, Boettcher cells, choroid plexus cells, pia arachnoid squamous epithelium Cells, pigmented ciliary epithelial cells, non-pigmented ciliary epithelial cells, corneal endothelial cells, PEG cells, airway Ciliated cells, oviductal ciliated cells, endometrial ciliated cells, reticulum testis ciliated cells, efferent ductal ciliated cells, ciliated Hairy ependymal cells, epidermal keratinocytes, epidermal basal cells, fingernail and toenail keratinocytes, nail Basal hair cells, medullary hair stem cells, cortical hair stem cells, cuticle hair stem cells, cuticle hair root sheath cells, Root sheath cells of the layer of Kusley, root sheath cells of the layer of Henle, outer root sheath cells, hair matrix cells, stratified squamous cell Surface epithelial cells of the skin, basal cells of the epithelium, urinary epithelial cells, inner hair cells of the organ of Corti of the ear, Outer hair cells of the organ of Corti, basal cells of the olfactory epithelium, cold-sensitive primary sensory neurons, heat-sensitive primary sensory neurons Sensory neurons, Merkel cells of the epidermis, olfactory receptor neurons, pain-sensitive primary sensory neurons, Photoreceptor rod cells, photoreceptor blue-sensitive cone cells, photoreceptor green-sensitive cone cells, photoreceptor Somatic red-sensitive cone cells, proprioceptive primary sensory neurons, tactile primary sensory neurons , type I carotid body cell, type II carotid body cell (blood pH sensor), type I vestibular apparatus of the ear Hair cells (acceleration and gravity), type II hair cells of the vestibular apparatus of the ear, type I taste bud cells Cholinergic Adrenergic neurons, peptidergic neurons, and the inner columnar cells of the organ of Corti Organ of Corti, outer column cells of the organ of Corti, inner supporting cells of the organ of Corti, outer supporting cells of the organ of Corti, border of the organ of Corti Boundary cells, Hensen's cells of the organ of Corti, vestibular apparatus supporting cells, taste bud supporting cells, olfactory epithelium supporting cells, Dewan cells, satellite cells, enteric glial cells, astrocytes, neurons, oligodendrocytes, spindle cells Pyramidal neurons, prelental epithelial cells, crystallin-containing lens fiber cells, hepatocytes, adipose cells Alveolar cells, white adipocytes, brown adipocytes, liver lipid cells, renal glomerular parietal cells, renal glomerular podocytes , brush border cells of the proximal renal tubule, cells of the thin part of the loop of Henle, cells of the distal renal tubule, cells of the renal collecting duct Cells, type I pneumocytes, pancreatic duct cells, smooth muscle duct cells, duct cells, intestinal brush border cells, exocrine gland streak cells Duct cells, gallbladder epithelial cells, non-ciliated cells of the efferent duct of the testis, epididymal principal cells, epididymal basal cells, Ameloblast epithelial cells, semilunar epithelial cells, interdental epithelial cells of the organ of Corti, loose connective tissue fibroblasts Cells, keratocytes, tendon fibroblasts, bone marrow reticular fibroblasts, non-epithelial fibroblasts, perithelium Cells, nucleus pulposus cells, cementoblasts / cementocytes entocytes, odontoblasts, odontocytes, hyaline Chondrocytes of cartilage, chondrocytes of fibrocartilage, chondrocytes of elastic cartilage, osteoblasts, osteocytes, osteoclasts alveolar cyst, osteocyte precursor cell, vitreous cell, stellate cell (ear), hepatic stellate cell (Ito cell), pancreatic stellate cell, red Muscle skeletal muscle cell, White skeletal muscle cell, Intermediate skeletal muscle cell, Muscle spindle nuclear bag cell, Muscle spindle nuclear chain cell , satellite cells, normal cardiomyocytes, nodal cardiomyocytes, Purkinje fiber cells, smooth muscle cells, iris Myoepithelial cells, myoepithelial cells of exocrine glands, reticulocytes, megakaryocytes, monocytes, connective tissue macrophages D. Epithelial Langerhans cells, dendritic cells, microglial cells, neutrophils, eosinophils, basophils , mast cells, helper T cells, suppressor T cells, cytotoxic T cells, natural killer Lar T cells, B cells, natural killer cells, melanocytes, retinal pigment epithelial cells, oocytes Follicle / oocyte, spermatid, spermatocyte, spermatogonia, sperm, follicle cell, Sertoli cell, thymus adjacent to cells selected from the group consisting of epithelial cells, interstitial kidney cells, or a mixture thereof. The cells may be fibroblasts derived from the tissue or the cells thereof.

[0078] As used within the context of this disclosure, fibroblasts and / or fibroblast-derived microorganisms The vehicle may contain one or more pharma- ceutically acceptable carriers in addition to the active ingredient for administration. The anti-cancer adjuvant of the present invention can be formulated by the above-mentioned method. Examples of excipients or diluents include lactose, dextrose, sucrose, sorbitol, etc. mannitol, xylitol, erythritol, maltitol, starch, arabic Rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, Chilled cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, hydroxybenzoic acid Methyl, propyl hydroxybenzoate, talc, magnesium stearate or mineral oil These include, but are not limited to:

[0079] When used as an oral or parenteral formulation, the fibroblasts are packaged in capsules, tablets, capsules, or capsule-shaped containers. Coating tablets, slow release tablets, granules, powders, syrups, suspensions, emulsions, Parenteral preparations may be administered as sterile aqueous solutions, non-aqueous solutions, or suppositories. Parenteral preparations can be intravenous, intravenous, suspension, emulsion and lyophilized preparations. Typically via intra-arterial, intraperitoneal, intramuscular, intrasternal, topical, rectal or intradermal routes. It can be administered.

[0080] Fibroblasts for oral administration contain diluents, preservatives, binders, lubricants, disintegrants, bulking agents, fillers, Pharmaceutically acceptable additives may typically include, but are not limited to, anti-inflammatory agents, stabilizers, and combinations thereof. The composition may be formulated with an acceptable carrier, such as a plasticizer, a colorant, a coloring agent, Components of the coating composition, which may include stabilizers and flow agents Examples of suitable coating materials include cellulose polymers, For example, cellulose acetate phthalate, hydroxypropyl cellulose, hydroxypropyl Methylcellulose, Hydroxypropyl Methylcellulose Phthalate and Hydroxypropyl propyl methylcellulose acetate succinate; polyvinyl acetate phthalate, a Acrylic acid polymers, acrylic acid copolymers, methacrylic resins, zein, shellac and Additionally, the coating material may contain, but is not limited to, a plasticizer. Typical carriers such as pigments, colorants, flow agents, stabilizers, pore formers and surfactants. Optional pharma- ceutically acceptable excipients may include diluents, binders, lubricants, These include, but are not limited to, lubricants, disintegrants, colorants, stabilizers or surfactants. .

[0081] As is known in the art, suitable formulations for compression of tablets or formation of beads and granules may be used. Diluents are usually required to increase the volume of a solid dosage form so that a desired particle size is provided. Suitable diluents include dicalcium phosphate dihydrate, calcium sulfate, lactose, , sucrose, mannitol, sorbitol, cellulose, microcrystalline cellulose, kaoli , sodium chloride, dry starch, hydrolyzed starch, pregelatinized starch, silicone Examples of suitable materials include titanium dioxide, titanium dioxide, magnesium aluminum silicate, or powdered sugar. Binders are used to impart cohesive properties to a solid dosage formulation. Therefore, it is necessary to ensure that the tablets or beads or granules remain unchanged after composition of the dosage form. Suitable binder materials include starch, pregelatinized starch, gelatin, Sugars (including sucrose, glucose, dextrose, lactose and sorbitol) , polyethylene glycols, waxes, natural and synthetic gums, e.g., acacia, tragacanthus and sodium alginate, cellulose (hydroxypropyl methylcellulose, hydroxypropyl cellulose, ethyl cellulose) and veegum, and Synthetic polymers, such as copolymers of acrylic acid and methacrylic acid, Polymers, methyl methacrylate copolymers, aminoalkyl methacrylate copolymers, poly Examples include, but are not limited to, acrylic acid / polymethacrylic acid and polyvinylpyrrolidone. In addition, lubricants are used to facilitate the preparation of tablets. Examples of the agent include magnesium stearate, calcium stearate, stearic acid, These include, but are not limited to, glycerol behenate, polyethylene glycol, talc and mineral oil. Disintegrants are used to facilitate the breakup or destruction of a dosage form after administration. These are usually starch, sodium starch glycolate, sodium Carboxymethyl starch, sodium carboxymethyl cellulose, hydroxypropyl cellulose, pregelatinized starch, clay, cellulose, arginine, rubber or cross-linked polypropylene Polymers include, but are not limited to, crosslinked PVP. Stabilizers are used to inhibit or retard drug decomposition reactions, including, for example, oxidative reactions. Suitable stabilizers include antioxidants, butyl hydroxytoluene, (BHT), ascorbic acid and its salts and esters; vitamin E, tocopherols sulfites such as sodium metabisulfite; cysteine ​​and its derivatives citric acid; propyl gallate; and butyl hydroxyanisole (BHA). These include, but are not limited to:

[0082] In some embodiments, oral dosage forms, such as capsules, tablets, liquids, and The suspension may be formulated for controlled release. For example, the suspension may be formulated to contain one or more compounds and optional and one or more further active ingredients of the formula (I) are nanoparticles, microparticles and combinations thereof. and encapsulated in soft or hard gelatin or non-gelatin capsules. or may be dispersed in a dispersion medium to form an oral suspension or syrup. These particles may be formed from a drug and a controlled release polymer or matrix. Alternatively, the drug particles may be coated with one or more controlled release coatings prior to incorporation into the finished dosage form. The composition may be coated with a penetrating agent.

[0083] In practicing the methods of the present disclosure, high plasma concentrations are obtained at the initiation of treatment to provide high plasma concentrations. It may be necessary to administer an initial dose of fibroblasts. This can be achieved by parenteral administration of the compound. Preparations for parenteral administration can be prepared as aqueous compositions using techniques known to those skilled in the art. Generally, such compositions can be prepared as injectable preparations, e.g., solutions or suspensions. in solid form, e.g., to prepare a solution or suspension by adding a reconstitution medium prior to injection; Microparticles or nanoparticles suitable for use in emulsions, e.g., water-in-oil (w / o) emulsions; emulsions or oil-in-water (o / w) emulsions and their microemulsions , liposomes or emulsomes. can be a solvent or dispersion medium, such as, for example, water, ethanol, one or more Polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol) Coal), oils (e.g., vegetable oils (e.g., peanut oil, corn oil, sesame oil, etc.) and Suitable fluidity includes, but is not limited to, the composition and combinations thereof. Use coating materials such as cellulose, and in the case of dispersions, maintain the required particle size. In addition, isotonic agents such as sugars or surfactants can be used to maintain the pH of the solution. The isotonicity agent can also include, but is not limited to, salts (e.g., sodium chloride). Solutions or preparations of the active compounds as free acids, free bases or pharma- ceutically acceptable salts are not permitted. The liquid or dispersion may be prepared by mixing water or another suitable solution with one or more pharma- ceutically acceptable excipients. The composition may be prepared in a solvent or dispersion medium. Examples of excipients include surfactants, dispersants, These include, but are not limited to, emulsifiers, pH modifiers, and combinations thereof. Suitable surfactants are anionic, cationic, amphoteric or nonionic surface active agents. Suitable anionic surfactants include carboxylate ions, sulfonate ions and and sulfate ion-containing surfactants. Examples of the antibacterial agent include sodium long-chain alkyl sulfonate and potassium long-chain alkyl sulfonate. Ammonium and long chain alkyl sulfonates, and sodium alkylaryl sulfonates Thorium, potassium alkylarylsulfonate and ammonium alkylarylsulfonate monium, for example, sodium dodecylbenzenesulfonate; dialkylsulfosuccinate Sodium, for example, sodium dodecylbenzenesulfonate; dialkyl sulfosuccinate sodium sulfosuccinate, for example, sodium bis-(2-ethylthiosuccinate); and alkyl sulfates, such as sodium lauryl sulfate. Cationic surfactants The agent may be a quaternary ammonium compound, such as benzalkonium chloride, benzethonium chloride, ammonium, cetrimonium bromide, stearyl dimethyl benzyl ammonium chloride, polyoxyethylene Non-ionic surfactants include, but are not limited to, ethylene and coconut amine. Examples of active agents include ethylene glycol monostearate, propylene glycol myristate, Recall, glyceryl monostearate, glyceryl stearate, polyglyceryl oleate Seryl-4, sorbitan acylate, sucrose acylate, PEG-150 laurate , PEG-400 monolaurate, Polyoxyethylene monolaurate, Polysorbate , Polyoxyethylene octylphenyl ether, PEG-1000 cetyl ether, poly Polyoxyethylene tridecyl ether, polypropylene glycol butyl ether, Xamer.RTM.401, stearoyl monoisopropanolamide and polyoxyethylene Examples of amphoteric surfactants include sodium N- Dodecyl-alanine, sodium N-lauryl-iminodipropionate, myristane These include foacetate, lauryl betaine and lauryl sulfobetaine. Not limited to. EXAMPLES

[0084] The following examples are included to demonstrate preferred embodiments of the disclosure. The techniques disclosed in the examples are techniques that the inventors have found to work well in the practice of the present disclosure. and therefore may be considered as a preferred mode for its implementation. However, those of ordinary skill in the art will recognize that, in light of this disclosure, Many variations can be made in the specific embodiments, and these variations are within the scope of the present disclosure. and recognize that without departing from the scope of the present invention, the same or similar results would be obtained. It is. Example 1: Systemic TNF-alpha inhibition by valproic acid-treated fibroblasts following PD-1 antibody Decrease in A

[0085] Saline (control), antibody against PD-l1 (PD-l1), or 5 μg / mL Balb / c fibroblasts cultured in 1000 mg / ml valproic acid for 24 hours ( 100,000 cells were injected intraperitoneally into the peritoneal cavity of Balb / c mice along with an antibody against PD-l1. Cells and antibodies were injected every other day. Serum TNF-alpha was measured using EL This was measured by ISA (Figure 1). Example 2: Reduction of lymphokine-activated killer (LAK) lethality by administration of fibroblasts under

[0086] 10,000 anti-CD3, anti-CD28 beads / ml with 100IU / ml LAK cells were cultured in IL-2 and C57 / BL6 splenocytes. Splenocytes were isolated by lysis of red blood cells with low-toxicity saline, and then cultured in phosphate-buffered saline. The cells were washed twice with water (PBS). The cells were cultured in a 5% CO2 atmosphere. The culture time was 96 hours, and the cell viability was 1.0 at the end of the culture. The reliability of Chromium was evaluated. 51 Cytotoxicity against K562 target cells using a release assay The cells were evaluated for activity. One million cells were administered per mouse.

[0087] Fibroblasts or bone marrow MSCs were obtained from SpinalCyte (CybroCell Cells were obtained from either Allcells or passage 3. As can be seen in Figure 2, the fibroblast-treated mice had significantly higher fibroblast-treated fibroblasts compared to MSCs. A substantial reduction in mortality was observed in mice treated with fibroblasts or MSCs. The injection was performed 4 hours after administration of AK cells.

[0088] Although the present disclosure and its advantages have been described in detail, the present invention is not limited to the disclosed subject matter, and the present invention is not limited to the disclosed subject matter. Various changes, substitutions, and alterations may be made herein without departing from the spirit and scope of the present invention. It should be understood that the scope of the present application is not limited to the processes described herein. The present disclosure is not limited to particular embodiments of the process, machine, manufacture, composition of matter, means, methods or steps. Those skilled in the art will be able to implement a method for making a substantially similar embodiment to the corresponding embodiment described herein. any currently existing or later developed technology that performs a function or achieves substantially the same result Any process, machine, article of manufacture, composition of matter, means, methods or steps utilized in accordance with the present disclosure are disclosed herein. It will be readily apparent from this disclosure that the present invention may be used in conjunction with the present invention. The present invention includes within its scope any such process, machine, manufacture, composition of matter, means, method or step. It is intended to be.

Claims

[Claim 1] 1. A method for reducing toxicity of a treatment to an individual, comprising administering to an individual an effective amount of fibroblasts and / or Fibroblast-derived microvesicles may be administered in conjunction with and / or prior to said treatment. and / or after administering said treatment to said individual, method.

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