Smallpox vaccine and stem cells for treatment of disease

Abstract

To provide methods and compositions for treating an inflammatory disease or infectious disease.SOLUTION: Provided is a method of treating an inflammatory or infectious disease in a subject in need of treatment by administering a poxvirus and stem cells to the subject, the disease being not cancer. The disease may be, for example, a chronic inflammatory disease (e.g., an autoimmune disease). Also provided is a composition comprising stem cells and a poxvirus, the poxvirus comprising a recombinant polynucleotide, and the recombinant polynucleotide encoding a therapeutic molecule.SELECTED DRAWING: None

Description

Technical Field

[0001]

Background Art

[0002]

[0001] Inflammatory diseases, such as autoimmune diseases, are caused by chronic inflammation in a subject. These diseases can cause symptoms ranging from mild discomfort to severe reactions and even death.

[0003]

[0002] Infectious diseases are caused by organisms such as bacteria, viruses, fungi, or parasites. Infections are often treated with antibiotics, antiviral drugs, antifungal drugs, antiprotozoal drugs, and anthelmintics, while pathogens are becoming increasingly resistant to these drugs. Other pathogens have no known treatment.

Summary of the Invention

Problems to be Solved by the Invention

[0004]

[0003] New methods for treating these diseases are needed.

Means for Solving the Problems

[0005]

[0004] A method and composition for treating a disease in a subject in need of treatment by administering a poxvirus and a stem cell to the subject are described herein, where the disease is not cancer.

[0006]

[0005] In one aspect, a method for treating a chronic inflammatory disease in a subject is provided. The method includes administering a poxvirus to the subject, where the disease is not cancer. In certain embodiments, the poxvirus is administered in a therapeutically effective amount, e.g., an amount sufficient to treat the chronic inflammatory disease.

[0007]

[0006] In one aspect, a method for treating an infectious disease and / or its symptoms in a subject is provided. The method includes administering a poxvirus to the subject. In certain embodiments, the poxvirus is administered in a therapeutically effective amount, e.g., an amount sufficient to treat the infectious disease and / or its symptoms. In certain embodiments, the disease is not cancer.

[0008]

[0007] In one aspect, a method for preventing an infectious disease and / or its symptoms in a subject is provided. The method includes administering a poxvirus to the subject. In certain embodiments, the poxvirus is administered in a therapeutically effective amount, e.g., an amount sufficient to prevent the infectious disease and / or its symptoms. In certain embodiments, the disease is not cancer.

[0009]

[0008] In one aspect, a method for preventing a cytokine storm in a subject is provided. The method includes administering a poxvirus to the subject. In certain embodiments, the poxvirus is administered in a therapeutically effective amount, e.g., an amount sufficient to prevent the cytokine storm. In certain embodiments, the cytokine storm is caused by an infection / infectious disease.

[0010]

[0009] In one aspect, a method for treating a disease characterized by chronic inflammation is provided. The method includes administering a poxvirus and stem cells to the subject, wherein the disease is not cancer. In certain embodiments, the poxvirus is administered in a therapeutically effective amount, e.g., an amount sufficient to treat the disease. In certain embodiments, the stem cell(s) is / are administered in a therapeutically effective amount, e.g., an amount sufficient to treat the disease. In certain embodiments, the poxvirus and the stem cells are present in the same (single) composition.

[0011] In one aspect, a method is provided for converting chronic inflammation into acute inflammation in a subject in need of such conversion. The method includes administering a poxvirus to the subject, where the disease is not cancer. In certain embodiments, the poxvirus is administered in a therapeutically effective amount, e.g., an amount sufficient to convert chronic inflammation into acute inflammation. In certain embodiments, the method further includes treating the acute inflammation. In certain embodiments, treating the acute inflammation includes administering to the subject a known treatment for acute inflammation.

[0012] In one aspect, a composition is provided that includes stem cells and optionally a poxvirus, where the poxvirus includes a recombinant polynucleotide that encodes a therapeutic molecule. In certain embodiments, the therapeutic molecule treats a chronic inflammatory disease. In certain embodiments, the therapeutic molecule is an anti-inflammatory molecule.

[0013] In certain embodiments, the disease is a chronic inflammatory disease. In certain embodiments, the chronic inflammatory disease is an autoimmune disease. In certain embodiments, the chronic inflammatory disease is asthma, chronic peptic ulcer, tuberculosis, arthritis, periodontitis, ulcerative colitis, Crohn's disease, rhinitis, active hepatitis, atherosclerosis, dermatitis, inflammatory bowel disease (IBD), systemic lupus, fibromyalgia, type I diabetes, psoriasis, multiple sclerosis, Addison's disease, Graves' disease, Sjogren's syndrome, Hashimoto's thyroiditis, myasthenia gravis, vasculitis, pernicious anemia, or celiac disease.

[0014]

[0013] In certain embodiments, the inflammatory disease is transplant rejection, Dupuytren's contracture, Peyronie's disease, periodontitis, endometriosis, hepatitis, glomerunephritis, atherscleroisis, cardiovascular disease, arthritis (e.g., osteoarthritis, rheumatoid arthritis, or psoriatic arthritis), inflammatory brain disease (including post-stroke encephalitis), atherosclerosis, traumatic injury, infection, and / or a shock state. In certain embodiments, the inflammatory disease is chronic obstructive pulmonary disease (COPD), e.g., emphysema, chronic bronchitis, or refractory (irreversible) asthma.

[0015]

[0014] In certain embodiments, the inflammatory disease is an enterocutaneous fistula, chronic radiation damage (which causes inflammatory tissue defects such as radiation cystitis or radiation enteritis), duodenal ulcer, or a chronic inflammatory disease of the central nervous system, e.g., post-stroke neuroinflammation, schizophrenia, autism, poisoning, chronic traumatic encephalopathy, or vaccine-induced neurotoxicity.

[0016]

[0015] In certain embodiments, the autoimmune disease is myasthenia gravis (MG), Hashimoto's thyroiditis, vasculitis, Graves' disease, psoriasis, chronic inflammatory demyelinating polyneuropathy (CIDP), Guillain - Barré syndrome, type 1 diabetes mellitus, lupus, multiple sclerosis, rheumatoid arthritis, Addison's disease, Sjogren's syndrome, celiac disease, myositis, ankylosing spondylitis, or scleroderma.

[0017]

[0016] In certain embodiments, the infectious disease is caused by bacteria, viruses, or fungi. In certain embodiments, the infectious disease is caused by viruses. In certain embodiments, the virus is rhinovirus, coronavirus, influenza, or respiratory syncytial virus. In certain embodiments, the coronavirus is severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).

[0018]

[0017] In certain embodiments, the inflammatory disease causes, or is capable of causing, a cytokine storm in the subject. In certain embodiments, the inflammatory disease causes, or is capable of causing, a cytokine storm in the subject. and is capable of causing a cytokine storm.

[0019]

[0018] In certain embodiments, the stem cells contain a recombinant polynucleotide. In certain embodiments, the recombinant polynucleotide encodes a therapeutic molecule. In certain embodiments, the poxvirus contains a recombinant polynucleotide. In certain embodiments, the recombinant polynucleotide encodes a therapeutic molecule.

[0020]

[0019] In certain embodiments, the therapeutic molecule treats the disease. In certain embodiments, the therapeutic molecule is a cytokine, a therapeutic antibody, a therapeutic fusion protein, RNA, a peptide, or a polypeptide. In certain embodiments, the cytokine is an anti-inflammatory cytokine. In certain embodiments, the cytokine is selected from interleukin (IL)-1 receptor antagonist, IL-4, IL-6, IL-10, IL-11, IL-13, IFN-alpha, and transforming growth factor-beta. In certain embodiments, the therapeutic molecule is selected from abatacept (Orencia), adalimumab (Humira), anakinra (Kineret), certolizumab (Cimzia), etanercept (Enbrel), golimumab (Simponi), infliximab (Remicade), ixekizumab (Taltz), natalizumab (Tysabri), rituximab (Rituxan), secukinumab (Cosentyx), tocilizumab (Actemra), ustekinumab (Stelara), vedolizumab (Entyvio), basiliximab (Simulect), daclizumab (Zinbryta), and muromonab (Orthoclone OKT3).

[0021]

[0020] In certain embodiments, the therapeutic molecule improves the treatment of the disease. For example, the therapeutic molecule can be a receptor that promotes the uptake of a therapeutic agent by cells expressing the therapeutic molecule. In another example, the therapeutic molecule can be an antigen recognized by a therapeutic agent. In another example, the therapeutic molecule can be an enzyme used by cells to produce a therapeutic agent (e.g., a steroid). In certain embodiments, the therapeutic agent is an agent that treats the disease.

[0022]

[0021] In certain embodiments, the method further comprises administering a therapeutic agent to the subject. In certain embodiments, the therapeutic agent is present in the same composition as the poxvirus and the stem cells. In certain embodiments, the therapeutic agent is administered separately from the poxvirus and the stem cells. In certain embodiments, the therapeutic agent is an agent that treats the disease. In certain embodiments, the therapeutic agent is selected from abatacept (Orencia), adalimumab (Humira), anakinra (Kineret), certolizumab (Cimzia), etanercept (Enbrel), golimumab (Simponi), infliximab (Remicade), ixekizumab (Taltz), natalizumab (Tysabri), rituximab (Rituxan), secukinumab (Cosentyx), tocilizumab (Actemra), ustekinumab (Stelara), vedolizumab (Entyvio), basiliximab (Simulect), daclizumab (Zinbryta), and muromonab (Orthoclone OKT3).

[0023]

[0022] In certain embodiments, the poxvirus, and optionally the stem cells, are administered to the subject by intravenous, intraperitoneal, intrathecal, intracardiac, intra-articular, intraventricular, intrapleural, intrasubstantial, or intraocular injection. In certain embodiments, the poxvirus, and optionally the stem cells, are administered directly to the region affected by the disease. In certain embodiments, the poxvirus, and optionally the stem cells, are administered by guided delivery, e.g., MRI-guided delivery.

[0024]

[0023] In certain embodiments, the stem cells are autologous to the subject. In certain embodiments, the stem cells are allogeneic to the subject. In certain embodiments, the subject is human. In certain embodiments, the subject is a non-human animal. In certain embodiments, the subject is a livestock animal. In certain embodiments, the subject is a pet.

[0025]

[0024] In certain embodiments, the poxvirus is vaccinia virus. In certain embodiments, the vaccinia virus is selected from the Dryvax strain, ACAM1000 strain, ACAM2000 strain, Lister strain, EM63 strain, LIVP strain, Tian Tan strain, Copenhagen strain, Western Reserve strain, Modified Vaccinia Ankara (MVA) strain, New York City Board of Health strain, Dalian strain, Ikeda strain, LC16M8 strain, Western Reserve Copenhagen strain, Tashkent strain, Tian Tan strain, Wyeth strain, IHD-J strain, and IHD-W strain, Brighton strain, Dalian I strain, and Connaught strain. In certain embodiments, the vaccinia virus is ACAM1000 or ACAM2000. In certain embodiments, the vaccinia virus is the New York City Board of Health strain. In certain embodiments, the poxvirus is an attenuated virus.

[0026]

[0025] In certain embodiments, the stem cells are selected from adult stem cells, embryonic stem cells, fetal stem cells, mesenchymal stem cells, neural stem cells, totipotent stem cells, pluripotent stem cells, multipotent stem cells, oligopotent stem cells, unipotent stem cells, adipose stromal cells, endothelial stem cells, induced pluripotent stem cells, bone marrow stem cells, umbilical cord blood stem cells, adult peripheral blood stem cells, myoblast stem cells, small juvenile stem cells, dermal fibroblast stem cells, and combinations thereof. In certain embodiments, the stem cells are derived from the subject to be treated with the composition.

[0027]

[0026] In certain embodiments, the stem cells are modified stem cells. In certain embodiments, the modification The stem cells express (are modified to express) a heterologous protein. In certain embodiments, the heterologous protein is a therapeutic molecule, a receptor that facilitates uptake of a therapeutic agent, an antigen recognized by a therapeutic agent, or an enzyme involved in the production of a therapeutic agent. In certain embodiments, the therapeutic agent is an agent that treats the disease.

DETAILED DESCRIPTION OF THE INVENTION

[0028]

[0027] After reading this description, it will be apparent to those skilled in the art how to implement the present invention in various alternative embodiments and alternative applications. However, not all of the various embodiments of the present invention will be described herein. The embodiments presented herein are presented by way of example only and should be understood to be non-limiting. Accordingly, this detailed description of the various alternative embodiments should not be construed as limiting the scope or breadth of the present invention as described below.

[0029]

[0028] Prior to disclosing and describing the present invention, it will be understood that the aspects described below are not limited to a particular composition, a method of preparing such a composition, or their use. This is because they can of course be diverse. Also, it will be understood that the terminology used herein is for the purpose of describing particular aspects only and is not intended to be limiting.

[0030]

[0029] The detailed description of the present invention is divided into various sections for the sole purpose of the reader's convenience, and the disclosure found in any section may be combined with the disclosure in another section. Headings or subheadings may be used in the specification for the reader's convenience and are not intended to affect the scope of the present invention. I. Definitions Unless otherwise noted, all technical and scientific terms used in this specification have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In this specification, as well as in the claims below, reference will be made to a number of terms that are defined to have the following meanings.

[0031] The technical terms used in this specification are for the purpose of describing particular embodiments only and are not intended to limit the present invention. As used in this specification, the singular forms "a", "an", and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise.

[0032] "Optional" or "optionally" means that the subsequent described event or circumstance may or may not occur, and that the description includes both the case where the event or circumstance occurs and the case where it does not.

[0033] The term "about", when used before a numerical representation that includes a range, such as temperature, time, amount, concentration, and the like, indicates an approximation that can vary by (+) or (-) 10%, 5%, 1%, or any sub-range or sub-value existing therebetween. Preferably, the term "about", when used with respect to the amount of a dosage, means that the dosage can vary by ±10%.

[0034]

[0034] "comprising" or "including" is intended to mean that the compositions and methods include the recited elements but do not exclude other things. "Consisting essentially of", when used to define compositions and methods, means excluding other elements that have any essential significance for the combination for the stated purpose. Thus, a composition consisting essentially of the elements as defined herein does not exclude other materials or steps that do not substantially affect the basic and novel feature(s) of the claimed invention. "Consisting of" means excluding other components and substantial method steps, other than trace elements. The embodiments defined by each of these transitional terms are within the scope of the present invention.

[0035]

[0035] The terms "disease" or "condition" refer to the existing or health state of a patient or subject that can be treated with the compounds or methods provided herein. The disease can be an autoimmune disease. The disease can be an inflammatory disease. The disease can be an infectious disease.

[0036]

[0036] As used herein, the term "inflammatory disease" refers to a disease or condition characterized by abnormal inflammation (e.g., increased levels of inflammation compared to controls such as healthy individuals without the disease). The term "chronic inflammatory disease" refers to an inflammatory disease that is persistent or recurrent. Examples of chronic inflammatory diseases include autoimmune diseases, arthritis, rheumatoid arthritis, psoriatic arthritis, juvenile idiopathic arthritis, multiple sclerosis, systemic lupus erythematosus (SLE), myasthenia gravis, juvenile-onset diabetes, type 1 diabetes mellitus, Guillain-Barré syndrome, Hashimoto's encephalopathy, Hashimoto's thyroiditis, ankylosing spondylitis, psoriasis, Sjögren's syndrome, vasculitis, glomerulonephritis, autoimmune thyroiditis, Behçet's disease, Crohn's disease, ulcerative colitis, bullous pemphigoid, sarcoidosis, ichthyosis, Graves' ophthalmopathy, inflammatory bowel disease, Addison's disease, vitiligo, asthma, allergic asthma, acne vulgaris, celiac disease, chronic prostatitis, inflammatory bowel disease, pelvic infectious disease, reperfusion injury, ischemia-reperfusion injury, stroke, sarcoidosis, transplant rejection, interstitial cystitis, atherosclerosis, scleroderma, and atopic dermatitis.

[0037]

[0037] In certain embodiments, the inflammatory disease is an enterocutaneous fistula, chronic radiation damage (which causes inflammatory tissue defects such as radiation cystitis or radiation enteritis), duodenal ulcer, or a chronic inflammatory disease of the central nervous system, such as neuroinflammation after stroke, schizophrenia, autism, poisoning, chronic traumatic encephalopathy, or vaccine-induced neurotoxicity.

[0038]

[0038] In certain embodiments, the inflammatory disease is transplant rejection, Dupuytren's contracture, Peyronie's disease, periodontitis, endometriosis, hepatitis, glomerulonephritis, arteriosclerosis, cardiovascular disease, arthritis (e.g., osteoarthritis, rheumatoid arthritis, or psoriatic arthritis), an inflammatory brain disease (including post-stroke encephalitis), atherosclerosis, traumatic injury, infection, and / or a shock state. In certain embodiments, the inflammatory disease is chronic obstructive pulmonary disease (COPD), such as emphysema, chronic bronchitis, or refractory (irreversible) asthma.

[0039] As used herein, the term "autoimmune disease" refers to a disease or condition in which the immune system of a subject has an abnormal immune response to a substance that does not normally induce an immune response in a healthy subject. Examples of autoimmune diseases that can be treated with the compounds, pharmaceutical compositions, or methods described herein include acute disseminated encephalomyelitis (ADEM), acute necrotizing hemorrhagic leukoencephalitis, Addison's disease, agammaglobulinemia, alopecia areata, amyloidosis, ankylosing spondylitis, anti-GBM / anti-TBM nephritis, antiphospholipid antibody syndrome (APS), autoimmune angioedema, autoimmune aplastic anemia, autoimmune autonomic neuropathy, autoimmune hepatitis, autoimmune hyperlipidemia, autoimmune immunodeficiency, autoimmune inner ear disease (AIED), autoimmune myocarditis, autoimmune oophoritis, autoimmune pancreatitis, autoimmune retinopathy, autoimmune thrombocytopenic purpura (ATP), autoimmune thyroid disease, autoimmune urticaria, axonal or neural type neuropathy, Baló disease, Behçet's disease, bullous pemphigoid, cardiomyopathy, Castleman disease, celiac disease, Chagas disease, chronic fatigue syndrome, chronic inflammatory demyelinating polyneuropathy (CIDP), chronic recurrent multifocal osteomyelitis (CRMO), Churg-Strauss syndrome, cicatricial pemphigoid / benign mucous membrane pemphigoid, Crohn's disease, Cogan syndrome, cold agglutinin disease, congenital heart block, coxsackievirus myocarditis, CREST disease, essential mixed cryoglobulinemia, demyelinating neuropathy, dermatitis herpetiformis, dermatomyositis, Devic's disease (neuromyelitis optica), discoid lupus, Dressler syndrome, endometriosis, eosinophilic esophagitis, eosinophilic fasciitis, erythema nodosum, experimental allergic encephalomyelitis, Evans syndrome, fibromyalgia, fibrosing alveolitis, giant cell arteritis (temporal arteritis), giant cell myocarditis, glomerulonephritis, Goodpasture syndrome, granulomatosis with polyangiitis (GPA) (formerly called Wegener's granulomatosis), Graves' disease, Guillain-Barré syndrome, Hashimoto's encephalopathy, Hashimoto's thyroiditis, hemolytic anemia, Henoch-Schönlein purpura, herpes gestationis, hypogammaglobulinemia, idiopathic thrombocytopenic purpura (ITP), IgA nephropathy, IgG4-related sclerosing disease, immunomodulatory lipoprotein, inclusion body myositis, interstitial cystitis, juvenile arthritis, juvenile diabetes (type 1 diabetes), Kawasaki syndrome, Lambert-Eaton syndrome, leukocytoclastic vasculitis, lichen planus, lichen sclerosus, ligneous conjunctivitis,Linear IgA disease (LAD), lupus (SLE), Lyme disease, chronic Meniere's disease, microscopic polyangiitis, mixed connective tissue disease (MCTD), Mooren ulcer, Mucosa-Herxheimer disease, multiple sclerosis, myasthenia gravis, myositis, narcolepsy, neuromyelitis optica (Devic's disease), neutropenia, ocular cicatricial pemphigoid, optic neuritis, relapsing rheumatism, PANDAS (pediatric autoimmune neuropsychiatric disorders associated with streptococcus), paraneoplastic cerebellar degeneration, paroxysmal nocturnal hemoglobinuria (PNH), Parry-Romberg syndrome, Personege-Turner syndrome, pars planitis (peripheral uveitis), pemphigus, peripheral neuropathy, perivenous encephalomyelitis, pernicious anemia, POEMS syndrome, polyarteritis nodosa, type I, type II, and type III polyglandular autoimmune syndrome, polymyalgia rheumatica, polymyositis, post-myocardial infarction syndrome, post-pericardiotomy syndrome, progesterone dermatitis, primary biliary cirrhosis, primary sclerosing cholangitis, psoriasis, psoriatic arthritis, idiopathic pulmonary fibrosis, pyoderma gangrenosum, erythroderma, Raynaud's phenomenon, reactive arthritis, reflex sympathetic dystrophy, Reiter's syndrome, relapsing polychondritis, restless legs syndrome, retroperitoneal fibrosis, rheumatic fever, rheumatoid arthritis, sarcoidosis, Schmidt syndrome, scleritis, scleroderma, Sjogren's syndrome, sperm & testicular autoimmunity, stiff-man syndrome, subacute bacterial endocarditis (SBE), Suzaku syndrome, sympathetic ophthalmia, Takayasu arteritis, temporal arteritis / giant cell arteritis, thrombotic thrombocytopenic purpura (TTP), Tolosa-Hunt syndrome, transverse myelitis, type 1 diabetes, ulcerative colitis, undifferentiated connective tissue disease (UCTD), uveitis, vasculitis, vesiculobullous skin disease, vitiligo, or Wegener's granulomatosis (i.e., granulomatosis with polyangiitis (GPA)).

[0040]

[0040] In certain embodiments, the inflammatory disease is an enteric fistula, chronic radiation damage (which causes inflammatory tissue defects such as radiation cystitis or radiation enteritis), duodenal ulcer, or a chronic inflammatory disease of the central nervous system, such as neuroinflammation after a stroke, schizophrenia, autism, poisoning, chronic traumatic encephalopathy, or vaccine-induced neurotoxicity.

[0041] In certain embodiments, the autoimmune disease is myasthenia gravis (MG), Hashimoto's thyroiditis, vasculitis, Graves' disease, psoriasis, chronic inflammatory demyelinating polyneuropathy (CIDP), Guillain-Barré, type 1 diabetes mellitus, lupus, multiple sclerosis, rheumatoid arthritis, Addison's disease, Sjögren's syndrome, celiac disease, myositis, ankylosing spondylitis, or scleroderma.

[0042] As used herein, the term "treating" or "treatment" refers to any indication of success of a therapy or improvement of an injury, disease, disorder or condition, including any objective or subjective parameter, e.g., alleviation; remission; decrease in symptoms or making the injury, disorder or condition more tolerable to the patient; slowing the rate of degeneration or decay; making the end point of degeneration less debilitating; or any indication of improving the physical or mental well-being of the patient. Treatment or amelioration of symptoms can be based on objective or subjective parameters including the results of a physical examination, neuropsychological tests, and / or psychiatric evaluations. The term "treating" and its conjugations can include preventing an injury, disorder, condition, or disease. In certain embodiments, treating is preventing. In certain embodiments, treating does not include preventing.

[0043]

[0043] Also, "treating" or "treatment", as used herein (and as is well understood in the art), broadly encompasses any approach for obtaining a beneficial or desired result in the condition of a subject, including clinical outcomes. Beneficial or desired results can include, but are not limited to, reduction or improvement of one or more symptoms or conditions, whether partial or total, and whether detectable or undetectable, decrease in the degree of a disease, stabilization of the disease condition (i.e., not worsening), prevention of the spread or dissemination of a disease, delay or slowing of disease progression, improvement or alleviation of the disease condition, reduction of disease recurrence, and remission. In other words, "treatment", as used herein, encompasses any cure, improvement, or prevention of a disease. Treatment can prevent a disease from occurring, inhibit its spread, reduce disease symptoms (e.g., eye pain, perception of blurred light around light, bloodshot eyes, very high eye pressure), completely or partially remove the underlying cause of the disease, shorten the duration of the disease, or a combination of these perspectives.

[0044]

[0044] "Treating" and "treatment", as used herein, encompass prophylactic treatment. The treatment method includes the step of administering to the subject a therapeutically effective amount of an active agent. The administering step may consist of a single administration or may include a series of administrations. The length of the treatment period depends on various factors such as the severity of the condition, the age of the patient, the concentration of the active agent, the activity of the composition used for treatment, or a combination thereof. Also, it will be understood that the effective dosage of the agent used for treatment or prophylaxis can increase or decrease over a particular treatment or prophylaxis regimen. Dose variations may occur and can be revealed by standard diagnostic assays known in the art. In some cases, chronic administration may be required. For example, the composition is administered to the subject in an amount sufficient to treat the patient and for a sufficient duration. In certain embodiments, treating or treatment is not prophylactic treatment.

[0045]

[0045] The term "prevent" refers to a reduction in the occurrence of disease symptoms in a patient. As noted above, prevention can be complete (undetectable symptoms) or partial, such that fewer symptoms are observed than would likely occur in the absence of treatment.

[0046]

[0046] "Patient," "subject," or "subject in need thereof" refers to any of the compounds provided herein. A patient refers to a living organism suffering from or prone to a disease or condition that can be treated by administration of such a pharmaceutical composition. Non-limiting examples include humans, other mammals, cows, rats, mice, dogs, monkeys, goats, sheep, dairy cows, deer, and other non-mammals. In some embodiments, the patient is a human. In some embodiments, the human is a pediatric patient. In some embodiments, the patient is a farm animal (e.g., goats, sheep, dairy cows, horses, etc.). In some embodiments, the patient is a pet, including, but not limited to, canines, felines, rodents (e.g., mice, rats, gerbils, hamsters, guinea pigs, chinchillas, etc.), rabbits, ferrets, etc.

[0047]

[0047] An "effective amount" is an amount of a compound sufficient to accomplish a stated purpose (e.g., to achieve the effect for which the compound is administered, to treat a disease, or to reduce one or more symptoms of a disease or condition) relative to the absence of the compound. An example of an "effective amount" is an amount sufficient to contribute to the treatment, prevention, or reduction of a symptom or symptoms of a disease, and an "effective amount" may also be referred to as a "therapeutically effective amount." A "reduction" of a symptom or symptoms (and grammatical equivalents of this phrase) means a decrease in the severity or frequency of the symptom or symptoms, or the elimination of the symptom or symptoms. The exact amount will depend on the purpose of the treatment and can be ascertained by one of ordinary skill in the art using known techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms (Vols. 1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999); and Remington: The Science and Practice of Pharmacy, 20th Edition, 2003, edited by Gennaro, Lippincott, Williams & Wilkins).

[0048]

[0048] As is well known in the art, a therapeutically effective amount for use in humans can also be determined from animal models. For example, a dosage for humans can be formulated to achieve a dosage known to be effective in animals. Dosages in humans can be adjusted by monitoring effectiveness and by adjusting the dosage up or down as described herein. Based on the methods described herein and other methods, adjusting the dosage to achieve maximum effectiveness in humans is well within the ability of one of ordinary skill in the art.

[0049]

[0049] As used herein, the term "therapeutically effective amount" refers to an amount of a therapeutic agent sufficient to ameliorate a disorder, as described above. For example, with respect to a given parameter, a therapeutically effective amount shows at least a 5%, 10%, 15%, 20%, 25%, 40%, 50%, 60%, 75%, 80%, 90%, or at least 100% increase or decrease. Therapeutic effectiveness can also be expressed as an "X-fold" increase or decrease. For example, a therapeutically effective amount can have at least a 1.2-fold, 1.5-fold, 2-fold, 5-fold, or higher effect than a control.

[0050]

[0050] The dosage can be varied according to the requirements of the patient and the composition used. The dosage administered to the patient should be sufficient to produce a beneficial therapeutic response in the patient over time, in the context of the present disclosure. The magnitude of the dosage is also determined by the presence, nature, and extent of any adverse side effects. Determining the dosage appropriate for a particular situation is within the skill of the physician. Generally, treatment is initiated at a lower, lesser amount of the dosage than the optimal dosage of the composition. Thereafter, the dosage is increased incrementally until the optimal effect is achieved in some circumstances. The amount and interval of the dosage can be adjusted individually to provide a dosage composition at an effective level for the particular clinical indication being treated. This provides a treatment regimen appropriate for the severity of the individual disease state.

[0051]

[0051] As used herein, the term "administering" means administering to a subject by oral administration, suppository, topical contact, intravenous, parenteral, intraperitoneal, intramuscular, intralesional, intrathecal, intracerebroventricular, intrapleural, parenchymal, intranasal, or subcutaneous administration, or implantation of a sustained release device, such as a mini-osmotic pump. Administration can be by any route, including parenteral and transmucosal (e.g., buccal, sublingual, palatal, gingival, nasal, vaginal, rectal, or transdermal). Examples of parenteral administration include intravenous, intramuscular, intraarterial, intradermal, subcutaneous, intraperitoneal, intracardiac, and intracranial. Other delivery modes include, but are not limited to, the use of liposomal formulations, intravenous infusion, etc. Administration also includes, for example, direct administration directly to the site of inflammation. Direct administration can be by guided delivery, such as magnetic resonance imaging (MRI) guided delivery. In some embodiments, administering does not include administering any active agent other than the recited active agents.

[0052] "Co-administered" is intended to mean that the compositions described herein are administered either simultaneously with, immediately prior to, or immediately following the administration of one or more additional therapies. The compositions provided herein may be administered to a patient either alone or co-administered. Co-administration is intended to include the simultaneous or sequential administration of the individual or combined (more than one composition) compositions. Thus, the preparation may also be combined with other active substances, if desired.

[0053] "Stem cells" are cells characterized by their ability to self-renew through mitotic cell division and their potential to differentiate into tissues or organs. Among mammalian stem cells, embryonic stem cells (ES cells) and somatic stem cells (e.g., HSC, iPSC) can be particularly excellent. Embryonic stem cells are present in the blastocyst and give rise to embryonic tissues, whereas somatic stem cells are present in adult tissues for the purpose of tissue regeneration and repair.

[0054]

[0054] The term "infection" or "infectious disease" refers to a disease or condition that can be caused by organisms such as bacteria, viruses, fungi, or any other pathogenic microbial pathogen. In certain embodiments, the infectious disease is caused by pathogenic bacteria. Pathogenic bacteria are bacteria that cause disease (e.g., in humans). In certain embodiments, the infectious disease is a bacteria-related disease (e.g., tuberculosis caused by Mycobacterium tuberculosis). Non-limiting examples of bacteria-related diseases include pneumonia that can be caused by bacteria such as Streptococcus or Pseudomonas, or food poisoning that can be caused by bacteria such as Shigella, Campylobacter, and Salmonella. Also included as bacteria-related diseases are tetanus, typhoid fever, diphtheria, syphilis, and Hansen's disease. In certain embodiments, the infectious disease is bacterial vaginitis (i.e., a bacterium that causes a change in the vaginal microbiota by overgrowth of bacteria that displace Lactobacilli species that maintain a healthy vaginal microbiota population) (e.g., yeast infection, or vaginal trichomonas); bacterial meningitis (i.e., bacterial inflammation of the meninges); bacterial pneumonia (i.e., bacterial infection of the lungs); urinary tract infection; bacterial gastroenteritis; or bacterial skin infection (e.g., impetigo, or cellulitis). In certain embodiments, the infectious disease is Campylobacter jejuni infection, Enterococcus faecalis infection, Haemophilus influenzae infection, Helicobacter pylori infection, Klebsiella pneumoniae infection, Legionella pneumophila infection, Neisseria gonorrhoeae infection, Neisseria meningitides infection, Staphylococcus aureus infection, Streptococcus pneumonia infection, Or it is an infection with Vibrio cholera. In certain embodiments, the infection is caused by a spirochete or borrelia, such as may be associated with Lyme disease.

[0055]

[0055] Terms such as "immune response" refer, in their ordinary sense, to the response by an organism that defends against a disease. The response can be enhanced by the innate immune system or by the adaptive immune system, as is well known in the art.

[0056]

[0056] The term "vaccine", as used herein, refers to any type of biological preparation that contributes to, or solicits, an active immune response against a particular disease or pathogen. Such biological preparations can include, but are not limited to, an antigen derived from an agent that causes a disease or a portion of an antigen derived from an agent that causes a disease. Such biological preparations can also exist in the form of a live attenuated preparation, including a live agent or pathogen that causes a disease, a debilitated agent or pathogen that causes a disease, or a modified agent or pathogen that causes a disease, or in the form of an inactivated or killed agent or pathogen that causes a disease. Further, alternative forms of such biological preparations include, but are not limited to, the form of a subunit, toxoid, conjugate, DNA, and recombinant vector, or any suitable form that can be developed or may be available in the future for soliciting an active immune response against them.

[0057]

[0057] In some embodiments, the term "vaccine" is used herein, but it should be noted that a vaccine does not necessarily provide significant immunity against smallpox (or any other pathogen), as long as it is effective against the disease as described herein. For example, a vaccine can be any immunogenic or infectious composition that treats a disease. In some cases, the term is used to identify a particular material or composition, and for example, it is not necessary to use it to identify the ability of the material or composition to provide immunity against smallpox. The virus can be derived from any strain of virus listed below, and elsewhere in this specification, including those that are not part of an approved or planned vaccine.

[0058]

[0058] As used herein, "virus" refers to any of a large group of entities referred to as viruses. Viruses typically contain a protein coat surrounding an RNA or DNA core of genetic material, do not contain a semipermeable membrane, and can grow and replicate only in living cells. Viruses for use in the methods provided herein include, but are not limited to, poxvirus, adenovirus, herpes simplex virus, Newcastle disease virus, vesicular stomatitis virus, mumps virus, influenza virus, measles virus, reovirus, human immunodeficiency virus (HIV), hantavirus, myxoma virus, cytomegalovirus (CMV), lentivirus, and any plant or insect virus.

[0059]

[0059] As used herein, "heterologous nucleic acid" refers to nucleic acid, DNA or RNA, that has been introduced into a virus or cell (or a progenitor of a cell). Such heterologous nucleic acid can include sequences related to genes and operable regulatory elements. For example, the heterologous nucleic acid can include a selectable marker gene, a suicide gene, or a gene that expresses a useful protein product that is not expressed endogenously or is expressed at a low endogenous level.

[0060] As used herein, the term "concurrently," when referring to administration of poxvirus and cells, refers to administration within 48 hours of each other. In some embodiments, the poxvirus and cells are administered within 36 hours of each other and within 24 hours of each other. within 12 hours of each other, within 10 hours of each other, within 8 hours of each other, within 6 hours of each other, within 4 hours of each other, within 2 hours of each other, within 1 hour of each other.

[0061]

[0061] The terms "autologous," "autologous cells," or "autologous transplant," when used herein in connection with cell transplantation, indicate that the donor and recipient of cells are the same individual. The terms "allogeneic," "allogeneic cells," or "allogeneic transplant," when used herein in connection with cell transplantation, indicate that the donor and recipient of cells are different individuals of the same species.

[0062]

[0062] The term "adipose tissue" as used herein refers to body adipose tissue, which is a loose connective tissue composed primarily of adipocytes. In addition to adipocytes, adipose tissue contains preadipocytes, fibroblasts, vascular endothelial cells, various immune cells, as well as the stromal vascular fraction (SVF) of cells that include regenerative stem cells.

[0063]

[0063] The term "adipose stem cells" or "adipose-derived adult stem cells (ADASC)" as used herein refers to stem cells that can be isolated from adipose tissue, which are pluripotent stem cells capable of differentiation into multiple cell types. Adipose stem cells have been shown to be comparable, if not superior, to bone marrow stem cells in terms of cell differentiation potential, angiogenesis, and anti-inflammatory effects.

[0064]

[0064] "Pharmaceutically acceptable excipients" and "pharmaceutically acceptable carriers" refer to substances that can be included in the compositions of the present disclosure to facilitate the administration of the active agent to the subject and / or its absorption by the subject without causing significantly harmful toxicological effects to the patient. Non-limiting examples of pharmaceutically acceptable excipients include water, NaCl, physiological saline, lactated Ringer's solution, standard sucrose, standard glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavors, salt solutions (e.g., Ringer's solution), alcohols, oils, gelatin, carbohydrates such as lactose, amylose or starch, fatty acid esters, hydroxymethylcellulose, polyvinylpyrrolidone, and pigments, among others. Such preparations may be sterilized and, if desired, may be mixed with auxiliaries such as lubricants, preservatives, stabilizers, wetting agents, emulsifying agents, salts that affect osmotic pressure, buffer solutions, colorants, and / or fragrances that do not react detrimentally with the compounds of the present disclosure. It will be understood by those skilled in the art that other pharmaceutical excipients are useful in the present disclosure. II. Poxvirus

[0065] Variola virus is the cause of smallpox. Unlike variola virus, vaccinia virus usually does not cause a systemic disease in immunocompetent individuals and is therefore used as a live vaccine for immunization against smallpox. Due to successful worldwide vaccination with vaccinia virus, smallpox has been eradicated as a natural disease. Routine smallpox vaccination has been interrupted for many years, except for people at higher risk of poxvirus infection (e.g., laboratory researchers). The United States discontinued routine childhood immunization against smallpox in 1972, but the use of smallpox vaccine is generally considered safe for use in children.

[0065]

[0066] Attenuated strains derived from pathogenic viruses can be used in the production of live vaccines. Non-limiting examples of virus strains that have been used as smallpox vaccines include the Lister strain (also known as Elstree), the New York City Board of Health strain (the "NYCBH strain"), the Dalian strain, the Ikeda strain, the LC16M8 strain, the Western Reserve (WR) strain, the Copenhagen strain, the Tashkent strain, the Tian Tan strain, the Wyeth strain, the IHD-J strain, and the IHD-W strain, the Brighton strain, the Ankara strain, the MVA strain, the Dalian I strain, the LIVP strain, the LC16MO strain, the LIVP strain, the WR 65-16 strain, the EM63 strain, and the Conn aught strain, but are not limited thereto. In some embodiments, the smallpox vaccine as disclosed herein is the attenuated New York City Board of Health (NYCBOH) strain of vaccinia virus. In some embodiments, the NYCBOH strain of vaccinia virus can be ATCC VR-118 or CJ-MVB-SPX.

[0066]

[0067] In some embodiments, the smallpox vaccine is not attenuated.

[0068] In some embodiments, the smallpox vaccine is selected from the Dryvax strain, the ACAM1000 strain, the ACAM2000 strain, the Lister strain, the EM63 strain, the LIVP strain, the Tian Tan strain, the Copenhagen strain, the Western Reserve strain, or the modified vaccinia Ankara (MVA) strain. In some embodiments, the smallpox vaccine does not lack any genes present in one or more of these strains.

[0067]

[0069] In some embodiments, the smallpox vaccine is a replication-competent virus. In some embodiments, the smallpox vaccine is replication-deficient. III. Stem Cells

[0070] The virus undergoes significant elimination and / or neutralization after systemic administration. Stem cells act as a vehicle to protect the disclosed smallpox vaccine from elements of humoral and cellular immunity in the bloodstream. See, for example, U.S. Patent No. 10,105,436, which is hereby incorporated by reference in its entirety.

[0068]

[0071] Accordingly, stem cells can be used as a vehicle for in vivo delivery of the poxvirus to the site of disease in a subject. In some embodiments, the poxvirus is mixed with stem cells to avoid the immune system eliminating the virus before it reaches the desired site. Accordingly, in some embodiments, a method of treating a disease in a subject, the method comprising administering to the subject the poxvirus in parallel with stem cells, is disclosed herein. In some embodiments, the poxvirus does not contain heterologous nucleic acids. In some embodiments, the method further comprises administering to the subject in parallel a genetically engineered virus. Some embodiments relate to methods of preparing a stem cell and poxvirus composition prior to administration.

[0069]

[0072] In some embodiments, the vehicle stem cells are autologous stem cells. In other embodiments, the vehicle stem cells are allogeneic or xenogeneic.

[0073] In some embodiments, the stem cells are selected from adult stem cells, embryonic stem cells, fetal stem cells, mesenchymal stem cells, neural stem cells, totipotent stem cells, pluripotent stem cells, multipotent stem cells, oligopotent stem cells, unipotent stem cells, adipose stromal cells, endothelial stem cells, induced pluripotent stem cells, bone marrow stem cells, umbilical cord blood stem cells, adult peripheral blood stem cells, myoblast stem cells, juvenile stem cells, dermal fibroblast stem cells, and combinations thereof. In some embodiments, the modified stem cells are umbilical cord-derived mesenchymal-like cells. In some embodiments, the umbilical cord-derived mesenchymal-like cells are IMMSTEM™ cells. In some embodiments, the stem cells are adipose stromal cells. One or more of the cells listed above can be specifically excluded from the compositions and methods of some embodiments.

[0070]

[0074] In some embodiments, the stem cells are modified. In particular, in some embodiments, the modified stem cells are adult stem cells (ASCs). In some embodiments, the modified stem cells are transformed with a viral vector. In some embodiments, the modified stem cells are transformed with a lentivirus or a retrovirus. In some embodiments, the modified stem cells are transformed with a recombinant virus. In some embodiments, the modified stem cells are transiently transfected with an artificial chromosome, a virus, or plasmid DNA. In some embodiments, the virus is an oncolytic virus. In some embodiments, the virus is a vaccinia virus. In some embodiments, the virus is a replication-competent oncolytic vaccinia virus (VACV). In some embodiments, the modified stem cells are capable of localizing to the site of disease in a subject. In some embodiments, the modified stem cells are autologous. In some embodiments, the modified stem cells are allogeneic.

[0071]

[0075] IMMSTEM™ cells are mesenchymal-like cells derived from the umbilical cord, and IMMSTEM™ cells possess pluripotent differentiation ability and are characterized by specific surface markers and growth factor production. IMMSTEM™ cells possess a number of advantages including ease of collection, a faster rate of proliferation, very low immunogenicity, and the ability to differentiate into tissues representing all three germ layer components compared to other stem cell sources. Compared to other mesenchymal stem cell (MSC) subtypes, IMMSTEM™ cells exhibit upregulation of anti-inflammatory and migratory abilities due to a "cytokine priming" step performed prior to administration. IMMSTEM™ cells are generated from the human umbilical cord obtained from women with normal full-term deliveries immediately after childbirth. To stimulate the stress response, the cells are cultured with interferon gamma for about 48 hours. In some embodiments, the culture with IFN gamma can be from 1 hour to 72 hours or any value or sub-range therein.

[0072]

[0076] In some embodiments, stem cells are infected with a poxvirus. Without being bound by theory, infection of the stem cells with the poxvirus is thought to enable further production of the poxvirus by cells that can target the diseased area.

[0073]

[0077] In some embodiments, stem cells are not infected with a poxvirus. In certain embodiments, stem cells that have not been infected with the poxvirus are engineered to express a therapeutic molecule. In some embodiments, stem cells infected with the poxvirus and stem cells that have not been infected and have been engineered to express a therapeutic molecule are administered to a subject. Without being bound by theory, the infected stem cells are thought to target the diseased area with the poxvirus, and the stem cells expressing the therapeutic molecule are thought to target the diseased area with the therapeutic molecule.

[0074]

[0078] U.S. Patent No. 10,105,436, which is hereby incorporated by reference in its entirety, describes poxviruses including smallpox vaccines that can be used in the methods and compositions described herein. Adipose stromal vascular cell group

[0079] In some embodiments, a method of treating a disease in a subject is disclosed, the method comprising administering to the subject a poxvirus in parallel with an adipose-derived stromal vascular fraction (SVF), wherein the disease is not cancer. In some embodiments, the adipose-derived SVF is autologous. In some embodiments, the adipose-derived SVF is administered to the subject within about 24 hours of the adipose tissue being harvested from the subject. In some embodiments, the adipose-derived SVF can be administered at any time after collection and up to about 48 hours after collection, or at any point or sub-range of time therebetween. In some embodiments, the smallpox vaccine is administered by intravenous, intraperitoneal, intrathecal, intracardiac, intra-articular, intraventricular, intrapleural, intrasubstantial, or intraocular injection or intradermal injection, or by any suitable method for delivering them.

[0075]

[0080] Adipose tissue is an alternative to bone marrow as a source of stem cells, including that: a) the extraction of adipose-derived cells is a simpler and less invasive procedure than bone marrow extraction; b) adipose tissue contains a higher content of mesenchymal stem cells (MSCs) compared to bone marrow; c) MSCs derived from adipose tissue do not decrease in number with aging and can thus function as an autologous cell source for all patients; and d) adipose tissue is also a source of a unique cell population, including endothelial cells, regulatory T cells, and monocytes / macrophages, in addition to the therapeutically capable MSCs, providing numerous benefits.

[0076]

[0081] MSCs have poor immunogenicity and possess immunomodulatory activity, a characteristic conserved among MSCs from various tissues. This weak immunogenicity is thought to permit the survival and activity of allogeneic MSCs when administered therapeutically.

[0077]

[0082] The SVF derived from the entire aspirated adipose tissue reduces the need for extensive processing of the internal cells, thereby minimizing the number of steps in which contamination can be introduced (Kurita et al., Plast. Reconstr. Surg. 2008, 121: 1033-1041; discussion 1042-1033; Yoshimura et al., Aesthetic Plast. Surg. 2008, 32: 48-55; discussion 56-47). The safety of adipose-derived cell administration is supported by autologous fat transplantation, a common practice in aesthetic surgery (Hang-Fu et al., Aesthetic Plast. Surg. 1995, 19: 427-437). Each of the above references is hereby incorporated by reference in its entirety.

[0078]

[0083] In some embodiments, adipose-derived SVF is obtained by means and knowledge known to those skilled in the art. In some embodiments, adipose-derived SVF is removed from a subject via a TIMEMACHINE (trademark) device. In some embodiments, adipose-derived SVF is removed from a subject using a 2.5 - 3 mM cannula. In some embodiments, one or more of the following devices can be utilized: PNC’s Multi Station, CHA Biotech Cha-Station, Cytori Celution 800 / CRS System, and Medi-Khan’s Lipokit with MaxStem.

[0079]

[0084] In some embodiments, the poxvirus and adipose-derived SVF are administered in parallel. In some embodiments, the poxvirus and adipose-derived SVF are administered simultaneously. In some embodiments, the poxvirus and adipose-derived SVF are administered simultaneously via one administration vehicle. In some embodiments, the poxvirus and adipose-derived SVF are administered simultaneously via one container, such as a syringe, by intravenous, intraperitoneal, intrathecal, intracerebroventricular, intra-articular, intraocular, intraventricular, intrapleural, intrasubstantial, or intradermal injection, or by any suitable method for delivering them.

[0080]

[0085] U.S. Patent No. 10,105,436, which is hereby incorporated by reference in its entirety, describes stem cells and SVF, including methods for obtaining / generating stem cells and SVF that can be used in the methods and compositions described herein. IV. Methods of Use Inflammatory Diseases

[0086] In one aspect, provided herein is a method of treating a chronic inflammatory disease in a subject in need of treatment, the method comprising administering to the subject a poxvirus (e.g., smallpox, e.g., smallpox vaccine) and stem cells, where the disease is not cancer. In certain embodiments, the chronic inflammatory disease is an autoimmune disease. In certain embodiments, the chronic inflammatory disease is asthma, chronic peptic ulcer, tuberculosis, arthritis, periodontitis, ulcerative colitis, Crohn's disease, rhinitis, active hepatitis, atherosclerosis, dermatitis, inflammatory bowel disease (IBD), systemic lupus, fibromyalgia, type 1 diabetes, psoriasis, multiple sclerosis, Addison's disease, Graves' disease, Sjogren's syndrome, Hashimoto's thyroiditis, myasthenia gravis, vasculitis, pernicious anemia, celiac disease. In certain embodiments, the inflammatory disease is transplant rejection, Dupuytren's contracture, Peyronie's disease, periodontitis, endometriosis, hepatitis, glomerulonephritis, arthritis (e.g., osteoarthritis, rheumatoid arthritis, or psoriatic arthritis), inflammatory brain disease (including post-stroke encephalitis), atherosclerosis. In certain embodiments, the autoimmune disease is myasthenia gravis (MG), Hashimoto's thyroiditis, vasculitis, Graves' disease, psoriasis, chronic inflammatory demyelinating polyneuropathy (CIDP), Guillain - Barré, type 1 diabetes mellitus, lupus, multiple sclerosis, rheumatoid arthritis, Addison's disease, Sjogren's syndrome, celiac disease, myositis, ankylosing spondylitis, or scleroderma.

[0081]

[0087] In certain embodiments, the inflammatory disease is enterocutaneous fistula, chronic radiation injury (which causes inflammatory tissue defects such as radiation cystitis or radiation enteritis), duodenal ulcer, chronic inflammatory disease of the central nervous system, e.g., post - stroke neuroinflammation, schizophrenia, autism, poisoning, chronic traumatic encephalopathy, or vaccine - induced neurotoxicity.

[0082]

[0088] In certain embodiments, the chronic inflammatory disease is transplant rejection, Dupuytren's contracture, Peyronie's disease, periodontitis, endometriosis, hepatitis, glomerulonephritis, arteriosclerosis, cardiovascular disease, arthritis (e.g., osteoarthritis, rheumatoid arthritis, or psoriatic arthritis), inflammatory brain disease (including post-stroke encephalitis), atherosclerosis, traumatic injury, infection, and / or shock state. In certain embodiments, the inflammatory disease is chronic obstructive pulmonary disease (COPD), e.g., emphysema, chronic bronchitis, or refractory (irreversible) asthma.

[0083]

[0089] In certain embodiments, the autoimmune disease is myasthenia gravis (MG), Hashimoto's thyroiditis, vasculitis, Graves' disease, psoriasis, chronic inflammatory demyelinating polyneuropathy (CIDP), Guillain - Barré, type 1 diabetes mellitus, lupus, multiple sclerosis, rheumatoid arthritis, Addison's disease, Sjögren's syndrome, celiac disease, myositis, ankylosing spondylitis, or scleroderma.

[0084]

[0090] In certain embodiments, the inflammatory disease is enterocutaneous fistula, chronic radiation damage (which causes inflammatory tissue defects such as radiation cystitis or radiation enteritis), duodenal ulcer, chronic inflammatory disease of the central nervous system, e.g., post - stroke neuroinflammation, schizophrenia, autism, poisoning, chronic traumatic encephalopathy, or vaccine - induced neurotoxicity.

[0085]

[0091] In certain embodiments, the disease is an infectious disease, a traumatic injury, and / or a shock state.

[0092] In certain embodiments, the therapeutic molecule (therapeutic agent) is administered to a subject. Preferably, the therapeutic molecule (therapeutic agent) treats the disease. The therapeutic molecule (therapeutic agent) can be administered as part of the poxvirus / stem cell composition and / or separately. When the therapeutic molecule (therapeutic agent) is administered as part of the poxvirus / stem cell composition, the therapeutic molecule (therapeutic agent) can be a separate component of the composition. Alternatively (or additionally), the therapeutic molecule may be expressed by the stem cells and / or encoded by the poxvirus.

[0086]

[0093] In certain embodiments, the therapeutic molecule (therapeutic agent) is a therapeutic agent listed in Table 1.

[0087]

Table 1

[0088]

[0094] In certain embodiments, the therapeutic molecule (therapeutic agent) is selected from abatacept (Orencia), adalimumab (Humira), anakinra (Kineret), certolizumab (Cimzia), etanercept (Enbrel), golimumab (Simponi), infliximab (Remicade), ixekizumab (Taltz), natalizumab (Tysabri), rituximab (Rituxan), secukinumab (Cosentyx), tocilizumab (Actemra), ustekinumab (Stelara), vedolizumab (Entyvio), basiliximab (Simulect), daclizumab (Zinbryta), and muromonab (Orthoclone OKT3).

[0089]

[0095] In certain embodiments, the therapeutic molecule comprises TGFβ, HGF, LIF, VEGF, EGF, BDNF, and / or NGF, or fragments thereof.

[0096] In certain embodiments, the therapeutic molecule is an antibiotic. Antibiotics are well known in the art. The antibiotic can be any antibiotic. A skilled clinician can determine which antibiotic should be used based on the type of infection and other standard determinations. Non-limiting examples of antibiotics are actinomycin, bacitracin, colistin, polymyxin B, gramicidins, polymyxins, bacitracins, glycopeptides, etc.

[0090]

[0097] In one aspect, a method is provided for converting chronic inflammation into acute inflammation in a subject in need of treatment. The method includes administering a poxvirus to the subject, where the disease is not cancer. In certain embodiments, the poxvirus is administered in a therapeutically effective amount, e.g., an amount sufficient to convert chronic inflammation into acute inflammation. In certain embodiments, the method further includes treating the acute inflammation. In certain embodiments, treating the acute inflammation includes administering to the subject a known treatment for acute inflammation.

[0091]

[0098] In certain embodiments, the disease is interstitial cystitis. Without being bound by theory, antiproliferative factor (APF) is thought to inhibit bladder cell proliferation by regulating cell adhesion proteins and growth factor production. In certain embodiments, the therapeutic agent expressed by the virus and / or stem cells is a polypeptide that blocks APF expression and / or activity. In certain embodiments, the therapeutic agent expressed by the virus and / or stem cells is a polypeptide that blocks NF-κB activity. In certain embodiments, the therapeutic agent expressed by the virus and / or stem cells is a polypeptide that blocks abnormal NF-κB activity. In certain embodiments, the therapeutic agent is HB-EGF or a fragment or variant thereof. See, for example, Kim et al., BJU Int. February 2009; 103(4):541-546, which is incorporated herein by reference in its entirety.

[0092]

[0099] In certain embodiments, the disease is atherosclerosis. Without being bound by theory, it is expected that induction of eNOS on the endothelial surface can alleviate atherosclerosis and even reverse plaque formation. In certain embodiments, the therapeutic agent expressed by the virus and / or stem cells is a polypeptide that induces eNOS.

[0093]

[0100] In certain embodiments, the infectious disease causes, or is capable of causing, a cytokine storm in a subject. In certain embodiments, the poxvirus, and optionally the administration of stem cells, treats the cytokine storm. Infectious disease

[0101] In certain aspects, a method of treating an infectious disease in a subject in need of treatment is provided. The method includes administering a poxvirus to the subject. In certain embodiments, the disease is not cancer.

[0094]

[0102] The infectious disease can be caused by any organism known to infect a subject. In certain embodiments, the infectious disease is caused by bacteria, viruses, parasites, protozoa, helminths, or fungi. In certain embodiments, the infectious disease is caused by bacteria. In certain embodiments, the infectious disease is caused by viruses. In certain embodiments, the infectious disease is caused by parasites. In certain embodiments, the infectious disease is caused by fungi. In certain embodiments, the infectious disease can be caused by one or more of the agents listed in U.S. Patent No. 8,715,677 and / or U.S. Patent Application Publication No. 2019 / 0381160, each of which is hereby incorporated by reference in its entirety.

[0095]

[0103] In certain embodiments, the virus is a virus that infects humans. In certain embodiments, the virus causes a respiratory infection, a gastrointestinal infection, food poisoning, a skin infection, or a sexually transmitted infection. In certain embodiments, the virus is a rhinovirus, a coronavirus, influenza, or a respiratory syncytial virus. In certain embodiments, the virus is a coronavirus. In certain embodiments, the coronavirus is severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).

[0096]

[0104] In certain embodiments, the stem cells are administered to a subject. The stem cells can be any type of stem cells as described herein, for example. In certain embodiments, the stem cells are mesenchymal stem cells. In certain embodiments, the stem cells are adipose stem cells.

[0097]

[0105] In certain embodiments, a therapeutic molecule (therapeutic agent) is administered to a subject. Preferably, the therapeutic molecule (therapeutic agent) treats a disease or a symptom of a disease. The therapeutic molecule (therapeutic agent) can be administered as part of the poxvirus / stem cell composition and / or separately. When the therapeutic molecule (therapeutic agent) is administered as part of the poxvirus / stem cell composition, the therapeutic molecule (therapeutic agent) can be a separate component of the composition. Alternatively (or further), the therapeutic molecule may be expressed by the stem cells and / or encoded by the poxvirus.

[0098]

[0106] In certain embodiments, the therapeutic molecule is an antibiotic. Antibiotics are well known in the art. The antibiotic can be any antibiotic. A skilled clinician can determine which antibiotic should be used based on the type of infection and other standard determinations. Non-limiting examples of antibiotics are actinomycin, bacitracin, colistin, polymyxin B, gramicidins, polymyxins, bacitracins, glycopeptides, etc.

[0099]

[0107] In certain embodiments, the therapeutic molecule is an antiviral molecule. In certain embodiments, the therapeutic molecule is an antifungal molecule. In certain embodiments, the therapeutic molecule is an antiparasitic molecule. In certain embodiments, the therapeutic molecule is a vaccine. In certain embodiments, the therapeutic molecules include molecules having anti-inflammatory and / or trophic activity. In certain embodiments, the therapeutic molecule is TGFβ, HGF, LIF, VEGF, EGF, BDNF, and / or NGF, or fragments thereof. In certain embodiments, the therapeutic molecule is a molecule that treats or prevents a cytokine storm, for example, leukemia inhibitory factor (LIF).

[0100]

[0108] In certain embodiments, the infectious disease causes or can cause a cytokine storm in a subject. In certain embodiments, the poxvirus, and optionally the administration of stem cells, treat the cytokine storm. Cancer patient

[0109] In certain aspects, provided is a method of treating or preventing an inflammatory disease or an infectious disease in a subject having cancer. Cancer patients are often immunocompromised, for example, as a result of treatment for cancer (chemotherapy, radiation, immunotherapy, etc.). Immunocompromised patients are more likely to be infected by pathogens that cause infectious diseases.

[0101]

[0110] In certain embodiments, the method includes administering a poxvirus to the subject. In certain embodiments, the inflammatory disease is treated. In certain embodiments, the inflammatory disease is prevented. In certain embodiments, the infectious disease is treated. In certain embodiments, the infectious disease is prevented. In certain embodiments, stem cells are administered. The subject can be treated using any method or composition described herein.

[0102]

[0111] Without being bound by theory, as described herein, administering a poxvirus, optionally together with stem cells, is thought to result in poxvirus infection of tumor cells in a patient. In certain embodiments, the poxvirus encodes a therapeutic molecule. In certain embodiments, the stem cells express a therapeutic molecule. In certain embodiments, the therapeutic molecule treats or prevents an inflammatory disease. In certain embodiments, the therapeutic molecule treats or prevents an infectious disease. In certain embodiments, the therapeutic molecule is a vaccine for a disease. In certain embodiments, the therapeutic molecule is an antibiotic. In certain embodiments, the therapeutic molecule is an antiviral molecule. In certain embodiments, the therapeutic molecule is an antifungal molecule. In certain embodiments, the therapeutic molecule is an antiparasitic molecule.

[0103]

[0112] In certain embodiments, infection of tumor cells with a poxvirus results in the expression of a therapeutic molecule by the tumor cells. Without being bound by theory, it is believed that the expression of the therapeutic molecule by the tumor cells results in an increase in the amount of therapeutic molecule delivered to the subject, enabling the treatment or prevention of disease. Administration

[0113] In certain embodiments, the poxvirus and / or stem cells are administered to a subject by intravenous, intraperitoneal, intrathecal, intracerebroventricular, intra-articular, intraventricular, intrapleural, intrasubstantial, or intraocular injection. In certain embodiments, the poxvirus, and optionally stem cells, are administered directly to the region affected by the disease. In certain embodiments, the poxvirus, and optionally stem cells, are administered by direct injection. In certain embodiments, the poxvirus, and optionally stem cells, are administered by MRI-guided delivery.

[0104]

[0114] In certain embodiments, the stem cells are autologous to the subject. That is, the stem cells are derived from the patient to be treated. For example, adipose-derived stromal vascular cell populations may be harvested from the patient, and stem cells may be collected from them or otherwise derived therefrom.

[0105]

[0115] In certain embodiments, the stem cells are allogeneic to the subject. In certain embodiments, the allogeneic stem cells are derived from a subject other than the patient. In certain embodiments, the allogeneic stem cells are derived from a cell line.

[0106]

[0116] In certain embodiments, the subject is human.

[0117] In some embodiments, the poxvirus and stem cells are administered simultaneously. In some embodiments, the poxvirus and stem cells are administered simultaneously via one administration vehicle. In some embodiments, the poxvirus and stem cells are administered simultaneously via one container, such as a syringe, by intravenous, intraperitoneal, intrathecal, intracerebroventricular, intra-articular, intraocular, intraventricular, intrapleural, intrasubstantial, or intradermal injection, or any suitable method for delivering them.

[0107]

[0118] The effective dosage for each of the treatment modalities disclosed herein can vary depending on a variety of factors including, but not limited to, the particular treatment, compound or pharmaceutical composition used, the mode of administration, the condition being treated, and / or the severity of the condition being treated. Thus, the dosage regimen of the combinations of the present invention is selected according to various factors including the route of administration and the renal and hepatic functions of the patient. A physician, clinician or veterinarian of ordinary skill in the art can readily determine and prescribe the effective amount of the single active ingredient required to prevent the progression of a condition, counteract the progression of a condition, or arrest the progression of a condition. Optimal precision to achieve a concentration of the active ingredient within a range that provides effectiveness without toxicity requires a regimen based on the kinetics of the availability of the active ingredient at the target site.

[0108]

[0119] The amount of poxvirus administered to an average-sized adult can be, for example, 1×10 2 ~1×10 10 plaque-forming units, 1×10 3 ~1×10 8 plaque-forming units, 1×10 4 ~1×10 6 plaque-forming units, or any value or subrange therebetween. As a specific example, about 2.5×10 5 plaque-forming units can be used.

[0109]

[0120] The embodiments described herein are not limited to vaccination or essentially vaccination, and it should be understood that they are also related to generating an immune response or a response to an antigen associated with a disease. While the terms "vaccine," "vaccination," or other similar terms are used for convenience, such embodiments are understood to be related to immune compositions, immunogenic compositions, immune response generation, immunization, etc. even when absolute prophylactic immunity is not required or generated. For example, embodiments referring to vaccination may also relate to creating or assisting in the creation of an immunogenic or immune response to an antigen, regardless of whether the response results in absolute eradication or immunization against the disease to be treated.

[0110]

[0121] U.S. Patent No. 10,105,436, which is hereby incorporated by reference in its entirety, describes methods of administering, making, storing, and using a composition comprising a poxvirus and stem cells that can be used in the methods and compositions described herein. V. Composition

[0122] In one aspect, a composition comprising a poxvirus (e.g., variola, e.g., variola vaccine) and optionally stem cells is provided herein, where the poxvirus comprises a recombinant polynucleotide that encodes a therapeutic molecule.

[0111]

[0123] In one aspect, a composition comprising a poxvirus (e.g., variola, e.g., variola vaccine) and stem cells is provided herein, where the stem cells comprise a recombinant polynucleotide that encodes a therapeutic molecule.

[0112]

[0124] In certain embodiments, the therapeutic molecule treats a chronic inflammatory disease. In certain embodiments, the therapeutic molecule treats an autoimmune disease. In certain embodiments, the therapeutic molecule treats an infectious disease, a traumatic injury, or a shock state. In certain embodiments, the therapeutic molecule is a cytokine, a therapeutic antibody, a therapeutic fusion protein, an antibiotic, RNA, a nucleotide, a peptide, or a polypeptide. In certain embodiments, the cytokine is selected from interleukin (IL)-1 receptor antagonist, IL-4, IL-6, IL-10, IL-11, IL-13, IFN-alpha, and transforming growth factor-beta.

[0113]

[0125] In one aspect, a composition comprising a poxvirus (e.g., variola, e.g., variola vaccine), optionally a stem cell, and a therapeutic agent is provided herein. In certain embodiments, the poxvirus comprises a recombinant polynucleotide that encodes a therapeutic molecule. In certain embodiments, the stem cell comprises a recombinant polynucleotide that encodes a therapeutic molecule.

[0114]

[0126] In certain embodiments, the therapeutic agent is a therapeutic agent listed in Table 1. In certain embodiments, the therapeutic agent is abatacept (Orencia), adalimumab (Humira), anakinra (Kineret), certolizumab (Cimzia), etanercept (Enbrel), golimumab (Simponi), infliximab (Remicade), ixekizumab (Taltz), natalizumab (Tysabri), rituximab (Rituxan), secukinumab (Cosentyx), tocilizumab (Actemra), ustekinumab (Stelara), vedolizumab (Entyvio), basiliximab ( Simulect), daclizumab (Zinbryta), or muromonab (Orthoclone OKT3).

[0115]

[0127] In certain embodiments, the RNA is antisense RNA, siRNA, an RNA vaccine, miRNA, or RNA interference (RNAi).

[0128] Antibiotics are well known in the art. A skilled clinician can determine which antibiotics should be used, as well as other standard determinations, based on the type of infection.

[0116]

[0129] The stem cells can be any stem cells, particularly stem cells as described herein.

[0130] The poxvirus can be any poxvirus, particularly a poxvirus as described herein. In certain embodiments, the poxvirus is an attenuated virus.

[0117]

[0131] Methods for preparing pharmaceutical compositions comprising the related treatments disclosed herein are known in the art and are apparent from known standard references such as Remington’s Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa., 18th Edition, (1990), which is hereby incorporated by reference in its entirety.

[0118]

[0132] In some embodiments, the compositions disclosed herein include a pharmaceutically acceptable carrier. As used herein, the term "pharmaceutically acceptable carrier" refers to solvents, diluents, preservatives, dispersion or suspension aids, isotonic agents, thickening or emulsifying agents, solid binders, and lubricants suitable for a particular dosage form. Those skilled in the art are aware of the various different carriers that can be used in formulating pharmaceutical compositions and are familiar with the techniques for their preparation (see Remington’s Pharmaceutical Sciences, edited by Gennaro, Mack Publishing Co., Easton, Pa., 1995, which is incorporated herein by reference in its entirety). Pharmaceutically acceptable carriers include, but are not limited to, Ringer's solution, isotonic saline, starch, potato starch, sugars, glucose, powdered tragacanth, malt, gelatin, talc, cellulose and its derivatives, ethyl cellulose, sodium carboxymethyl cellulose, cellulose acetate excipients, cocoa butter, suppository wax, agar, alginic acid, oils, cottonseed oil, peanut oil, safflower oil, sesame oil, olive oil, soybean oil, corn oil, glycols, propylene glycol, esters, ethyl laurate, ethyl oleate, buffer agents, aluminum hydroxide, magnesium hydroxide, phosphate buffer solutions, pyrogen-free water, ethyl alcohol, other non-toxic compatible lubricants, sodium lauryl sulfate, magnesium stearate, coloring agents, release agents, coating agents, sweetening agents, flavoring agents, and fragrances. Pharmaceutically acceptable carriers may also include preservatives and antioxidants. One or more of the foregoing materials may be specifically excluded from the compositions and methods of some embodiments.

[0119]

[0133] Compositions disclosed herein that include poxvirus may include an adjuvant. Optionally, one or more compounds having adjuvant activity may be included in the composition. An adjuvant is a non-specific stimulator of the immune system that enhances the host immune response to a vaccine. Examples of adjuvants known in the art are Freund's complete and incomplete adjuvants, vitamin E, nonionic block polymers, muramyl dipeptide, ISCOM (immunostimulating complex), saponin, mineral oil, vegetable oil, and carbopol. Adjuvants particularly suitable for mucosal application are, for example, Escherichia coli (E. coli) heat-labile toxin (LT) or cholera toxin (CT). Other suitable adjuvants are, for example, aluminum hydroxide, aluminum phosphate, or aluminum oxide, oil-emulsions (e.g., Bayol F® or Marcol 52®, saponin, or vitamin E solubilizing solution). One or more of the above-described materials may be specifically excluded from the compositions and methods of some embodiments.

[0120]

[0134] U.S. Patent No. 10,105,436, which is hereby incorporated by reference in its entirety, describes methods of administering, making, storing, and using compositions comprising poxvirus and stem cells that can be used in the methods and compositions described herein.

[0121]

[0135] The examples and embodiments described herein are for illustrative purposes only, and various modifications or changes in light of them will be suggested to those skilled in the art and should be included within the spirit and scope of this application, as well as the appended claims. All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety for all purposes.

Examples

[0122] Example 1: Preparation of ACAM2000 vaccine

[0136] The vaccine vial should be removed from cold storage, brought to room temperature, and then reconstituted. Remove the flip cap seals from the vaccine vial and the diluent vial, wipe each rubber stopper with an isopropyl alcohol swab, and allow to dry completely.

[0123]

[0137] Using aseptic technique and a sterile 1 mL syringe fitted with a 25-gauge x 5 / 8” needle, transfer 0.3 mL of diluent to the vaccine vial. Gently swirl the vial, preferably without touching the product to the rubber stopper, to mix. The reconstituted vaccine should be a clear to slightly turbid, colorless to pale yellow liquid free of foreign matter. The reconstituted vaccine is visually verified for particulate matter and discoloration prior to administration. If particulate matter or discoloration is observed, the vaccine is not used and the vial is disposed of safely.

[0124]

[0138] After reconstitution of the lyophilized preparation, each vial contains approximately 2.5 - 12.5 x 10 7 plaque-forming units (pfu) of live vaccinia virus. After reconstitution, ACAM2000 vaccine can be used within 6 to 8 hours if held at room temperature (20°C - 25°C, 68°F - 77°F). Unused reconstituted ACAM2000 vaccine may be stored in a refrigerator (2°C - 8°C, 36°F - 46°F) for up to 30 days, after which it should be discarded as biohazardous material. Persons preparing and administering the vaccine should wear surgical or protective gloves to avoid contact of the vaccine with skin, eyes, or mucous membranes. The vaccine vial, its stopper, the diluent syringe, the vent needle used for reconstitution, the needle used for administration, and any material that has come into contact with the vaccine should be discarded in a leak-proof, non-punctured biohazard container. Subsequently, these containers should be disposed of appropriately.

[0125]

[0139] For vaccine application, gently swirl and mix the vaccine vial, preferably without attaching the product to the rubber stopper. Using aseptic technique and a sterile 1 mL syringe fitted with a 25-gauge × 1.5875 cm (5 / 8 inch) needle, transfer the entire contents of the vial to a labeled 20 mL (20 cc) syringe containing the SVF fraction. Gently swirl the SVF vaccine mixture to mix well and incubate at 37 °C for 2 to 4 hours. Example 2: Recovery and preparation of adipose stromal vascular cell group

[0140] Administer a local anesthetic composed of lidocaine 0.5%, epinephrine (1:400,000), and HCO3 (8.4%) titrated to pH 7.4 (generally 5 mL (5 cc) of HCO3 in a total volume of 60 mL (60 cc)) to the patient and apply a sterile preparation. Next, administer a liposuction procedure to the patient using, for example, a TIME-MACHINE (trademark) device, a fat processing unit knit (syringe) and a 2.5 mm to 3 mm cannula. Apply bacitracin ointment and a band-aid and secure them over the wound with a compression dressing.

[0126]

[0141] Prepare SVF (ADSC) in a closed system according to the following protocol: a. Collect fat into a 60 mL (60 cc) single-use sterile fat processing syringe using a TIME-MACHINE (registered trademark).

[0127] b. Centrifuge the fat at 2800 rpm for 3 minutes. c. Remove free fatty acids and debris (local / blood) through a TP-109 closed system. d. Transfer 25 mL (25 cc) of concentrated fat to an SVF processing syringe.

[0128] e. Add 25 mL (25 cc) of pre-warmed (38 °C) T-MAX (registered trademark) Time Machine Accelerator (GMP-grade collagenase; Roche), containing 12.5 Wunsch units.

[0129] f. Incubate at 38 °C for 30 to 45 minutes. g. Centrifuge at 200×g for 4 minutes. h. Remove the supernatant liquid, except for 3 mL (3 cc) to 10 mL (10 cc) at the bottom.

[0130] i. Add 50 mL (50 cc) of D5LR as a washing solution to remove the collagenase residue and centrifuge at 200×g for 4 minutes. j. Repeat two more times for a total of three washes.

[0131] k. Leave 3 mL (3 cc) to 10 mL (10 cc) of the pellet collection and remove all of the supernatant liquid - this is the stromal vascular fraction cells. l. Filter the SVF through a 100 micron filter into a 20 mL (20 cc) syringe. m. Collect the SVF sample, identify it with respect to cell count and viability, and confirm that there is no aggregation or debris.

[0132] n. Aliquot each cell suspension for endotoxin experiments and sterility staining. The SVF is shipped for injection only after confirmation of endotoxin assay results of less than or equal to 5 EU / kg / hour and Gram stain negative results.

[0133] o. Resuspend the cells in 20 ml of saline. Aspirate the cell suspension into a syringe through an 18-gauge needle for injection. A maximum of 100 million viable cells are used for injection. p. Subsequently, place the syringe into a sealed specimen bag labeled with the patient's name and medical record number for transport to the injection treatment room. Example 3: Administration of adipose SVF with smallpox vaccine Development method:

[0142] Intravenous: A non-proliferating autologous stromal vascular fraction (SVF) extracted from a maximum of 500 ml of liposuction tissue, purified by collagenase digestion and a series of washing steps, containing a maximum of 100 million cells, is incubated with vaccinia virus and delivered by intravenous injection with a volume of 20 mL.

Claims

1. 11. A method of treating a chronic inflammatory disease in a subject in need thereof, comprising administering to the subject a poxvirus, wherein the disease is not cancer.

2. 1. A method of treating a disease characterized by chronic inflammation in a subject in need thereof, comprising administering to the subject a poxvirus, wherein the disease is not cancer.

3. 11. A method of converting chronic inflammation to acute inflammation, comprising administering to a subject a poxvirus, wherein the disease is not cancer.

4. 11. A method of treating an infectious disease in a subject in need thereof, comprising administering to the subject a poxvirus, wherein the disease is not cancer.

5. A method of treating a cytokine storm in a subject in need thereof, comprising administering to the subject a poxvirus, wherein the disease is not cancer.

6. The method of claim 4 , wherein the subject has an inflammatory disease.

7. The method of claim 4 , wherein the subject has an infectious disease.

8. 2. A method of treating or preventing an inflammatory or infectious disease in a subject having cancer, comprising administering to the subject a poxvirus.

9. The method of any one of claims 1 to 7, wherein the stem cells are administered together with a poxvirus.

10. 9. The method of any one of claims 1, 2, 7 and 8, wherein the chronic inflammatory disease is an autoimmune disease.

11. 10. The method according to any one of claims 1 to 9, wherein the chronic inflammatory disease is selected from asthma, chronic peptic ulcer, tuberculosis, arthritis, periodontitis, ulcerative colitis, Crohn's disease, sinusitis, active hepatitis, atherosclerosis, dermatitis, inflammatory bowel disease (IBS), systemic lupus, fibromyalgia, type I diabetes, psoriasis, multiple sclerosis, Addison's disease, Graves' disease, Sjogren's syndrome, Hashimoto's thyroiditis, myasthenia gravis, vasculitis, pernicious anemia, or celiac disease.

12. 10. The method of claim 9, wherein the autoimmune disease is myasthenia gravis (MG), Hashimoto's thyroiditis, vasculitis, Graves' disease, psoriasis, chronic inflammatory demyelinating polyneuropathy (CIDP), Guillain-Barre, type 1 diabetes mellitus, lupus, multiple sclerosis, rheumatoid arthritis, Addison's disease, Sjogren's syndrome, celiac disease, myositis, ankylosing spondylitis, or scleroderma.

13. 10. The method of any one of claims 1 to 9, wherein the chronic inflammatory disease is transplant rejection, Dupuytren's contracture, Peyronie's disease, periodontitis, endometriosis, hepatitis, glomerulonephritis, arteriosclerosis, cardiovascular disease, arthritis (e.g. osteoarthritis, rheumatoid arthritis, or psoriatic arthritis), inflammatory brain disease (including post-stroke encephalitis), atherosclerosis, traumatic injury, infection, chronic obstructive pulmonary disease (COPD), and / or shock states.

14. The inflammatory disease may be intestinal fistula, chronic radiation injury (which causes inflammatory tissue defects such as radiation cystitis or radiation enteritis), duodenal ulcer, or chronic inflammatory diseases of the central nervous system, such as neuroinflammation after stroke, schizophrenia, autism, poisoning, chronic traumatic encephalopathy, or vaccine-induced The method according to any one of claims 1 to 9, wherein the neurotoxicity is

15. 13. The method of claim 12, wherein the COPD is emphysema, chronic bronchitis, or refractory (irreversible) asthma.

16. The method according to any one of claims 3 to 8, wherein the infectious disease is caused by a bacterium, a virus, or a fungus.

17. 16. The method of claim 15, wherein the infectious disease is caused by a virus.

18. 17. The method of claim 16, wherein the virus is a rhinovirus, coronavirus, influenza, or respiratory syncytial virus.

19. The coronavirus is severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).

20. The method of claim 18, wherein:

20. The method according to any one of claims 1 to 19, wherein the poxvirus is a vaccinia virus.

21. 21. The method of claim 20, wherein the vaccinia virus is selected from Dryvax, ACAM1000, ACAM2000, Lister, EM63, LIVP, Tian Tan, Copenhagen, Western Reserve, Modified Vaccinia Ankara (MVA), New York City Board of Health, Dalian, Ikeda, LC16M8, Western Reserve Copenhagen, Tashkent, Tian Tan, Wyeth, IHD-J, and IHD-W, Brighton, Dalian I, and Connaught strains.

22. 22. The method of claim 21, wherein the vaccinia virus is ACAM1000 or ACAM2000.

23. 22. The method of claim 21, wherein the vaccinia virus is the New York City Board of Health strain.

24. The method according to any one of claims 1 to 22, wherein the poxvirus is an attenuated virus.

25. The method of any one of claims 1 to 24, further comprising administering stem cells to a subject.

26. 26. The method of claim 25, wherein the stem cells comprise a recombinant polynucleotide, said recombinant polynucleotide encoding a therapeutic molecule.

27. The method of any one of claims 1 to 26, wherein the poxvirus comprises a recombinant polynucleotide, said recombinant polynucleotide encoding a therapeutic molecule.

28. 28. The method of claim 26 or 27, wherein the therapeutic molecule treats a disease.

29. 29. The method according to any one of claims 26 to 28, wherein the therapeutic molecule is a cytokine, a therapeutic antibody, a therapeutic fusion protein, an antibiotic, RNA, a receptor that facilitates uptake of a therapeutic agent, an antigen recognized by a therapeutic agent, or an enzyme used by the cell to produce a therapeutic agent. How to.

30. The therapeutic molecule is selected from an anti-TNF antibody, a T cell receptor-directed antibody, an IL-2 receptor-directed antibody, and an interferon, and the therapeutic molecule is selected from abatacept (Orencia), adalimumab (Humira), anakinra (Kineret), certolizumab (Cimzia), etanercept (Enbrel), golimumab (Simponi), infliximab (Remicil), 30. The method of any one of claims 26-29, wherein the therapeutic agent is selected from the group consisting of ade, ixekizumab (Taltz), natalizumab (Tysabri), rituximab (Rituxan), secukinumab (Cosentyx), tocilizumab (Actemra), ustekinumab (Stellara), vedolizumab (Entyvio), basiliximab (Simulect), daclizumab (Zinbryta), and muromonab (Orthoclone OKT3).

31. The method of any one of claims 1 to 30, further comprising administering a therapeutic agent to the subject.

32. 32. The method of claim 31, wherein the therapeutic agent is an agent that treats a chronic inflammatory disease.

33. 33. The method of claim 31 or 32, wherein the therapeutic agent is selected from 5-aminosalicylic acid, corticosteroids, azathioprine, mercaptopurine, cyclosporine, bronchodilators, roflumilast, statins, fibrates, beta blockers, ACE inhibitors, diuretics, aspirin, calcium channel blockers, collagenase, verapamil, topical antiseptics, penicillins, antibiotics, hormones, hepatitis A vaccine, hepatitis B vaccine, immunosuppressants, cyclophosphamide, NSAIDs, analgesics, anesthetics, steroids, proton pump inhibitors, antivirals, anticonvulsants, anticoagulants, antihypertensives, and epinephrine.

34. The therapeutic agents include abatacept (Orencia), adalimumab (Humira), anakinra (Kineret), certolizumab (Cimzia), etanercept (Enbrel), golimumab (Simponi), infliximab (Remicade), ixekizumab (Taltz), natalizumab (Tysabri), rituximab (Rituxan), secukinumab (Cosentyx), tocilizumab (Actemra), ustekinumab (Stellara), vedolizumab (Entyvio), basiliximab (Simulect), daclizumab (Zinbryta), and muromonab (Orthoclone).

33. The method of claim 31 or 32, wherein the compound is selected from the group consisting of methyl ketone, ...

35. 32. The method of claim 31, wherein the therapeutic agent is an agent that treats a viral infection or a symptom thereof.

36. 36. The method of any one of claims 25 to 35, wherein the stem cells are selected from adult stem cells, embryonic stem cells, fetal stem cells, mesenchymal stem cells, neural stem cells, totipotent stem cells, pluripotent stem cells, multipotent stem cells, oligopotent stem cells, unipotent stem cells, adipose stromal cells, endothelial stem cells, induced pluripotent stem cells, bone marrow stem cells, umbilical cord blood stem cells, adult peripheral blood stem cells, myoblast stem cells, juvenile stem cells, skin fibroblast stem cells, and combinations thereof.

37. 37. The method of claim 36, wherein the stem cells are adipose stem cells.

38. The method of any one of claims 25 to 37, wherein the stem cell is or is derived from a stem cell lineage.

39. The method of any one of claims 25 to 38, wherein the stem cells are modified stem cells.

40. 40. The method of claim 39, wherein the modified stem cell expresses a heterologous protein.

41. 41. The method of claim 40, wherein the heterologous protein is a therapeutic molecule.

42. 42. The method of claim 41, wherein the therapeutic molecule treats the disease.

43. 43. The method of claim 41 or 42, wherein the therapeutic molecule is a cytokine, a therapeutic antibody, a therapeutic fusion protein, an antibiotic, RNA, a receptor that facilitates uptake of a therapeutic agent, an antigen recognized by a therapeutic agent, or an enzyme used by the cell to produce a therapeutic agent.

44. The therapeutic molecule is selected from an anti-TNF antibody, a T cell receptor-directed antibody, an IL-2 receptor-directed antibody, and an interferon, and the therapeutic molecule is selected from abatacept (Orencia), adalimumab (Humira), anakinra (Kineret), certolizumab (Cimzia), etanercept (Enbrel), golimumab (Simponi), infliximab (Remicil), 44. The method of any one of claims 41 to 43, wherein the therapeutic agent is selected from the group consisting of ade, ixekizumab (Taltz), natalizumab (Tysabri), rituximab (Rituxan), secukinumab (Cosentyx), tocilizumab (Actemra), ustekinumab (Stellara), vedolizumab (Entyvio), basiliximab (Simulect), daclizumab (Zinbryta), and muromonab (Orthoclone OKT3).

45. 45. The method of any one of claims 1 to 44, wherein the poxvirus and / or stem cells are administered to the subject by intravenous, intraperitoneal, intrathecal, intracerebroventricular, intrapleural, intraparenchymal, intraventricular, intraarticular, or intraocular injection.

46. The method according to any one of claims 1 to 44, wherein the poxvirus and / or the stem cells are administered directly to the diseased area.

47. 47. The method of claim 46, wherein the poxvirus and / or stem cells are administered by MRI-guided delivery.

48. 48. The method of any one of claims 1 to 47, wherein the stem cells are autologous to the subject.

49. 49. The method of any one of claims 1 to 48, wherein the stem cells are allogeneic to the subject.

50. The method of any one of claims 1 to 49, wherein the subject is a human.

51. 50. The method of any one of claims 1 to 49, wherein the subject is a livestock animal.

52. The method of any one of claims 1 to 49, wherein the subject is a pet.

53. 53. The method of claim 52, wherein the subject is a canine.

54. A composition comprising stem cells and a poxvirus, wherein the poxvirus comprises a recombinant polynucleotide, said recombinant polynucleotide encoding a therapeutic molecule.

55. The composition of claim 54, wherein the therapeutic molecule treats an inflammatory disease.

56. 56. The composition of claim 54 or 55, wherein the therapeutic molecule is a cytokine, a therapeutic antibody, a therapeutic fusion protein, RNA, an antibiotic, a receptor that facilitates uptake of a therapeutic agent, an antigen recognized by a therapeutic agent, or an enzyme used by the cell to produce a therapeutic agent.

57. 56. The composition of claim 54 or 55, wherein the therapeutic molecule is selected from an anti-TNF antibody, a T cell receptor-directed antibody, an IL-2 receptor-directed antibody, and an interferon.

58. The therapeutic molecules include abatacept (Orencia), adalimumab (Humira), anakinra (Kineret), certolizumab (Cimzia), etanercept (Enbrel), golimumab (Simponi), infliximab (Remicade), ixekizumab (Taltz), natalizumab (Tysabri), rituximab (Rituxan), secukinumab (Cosentyx), tocilizumab (Actemra), ustekinumab (Stellara), vedolizumab (Entyvio), basiliximab (Simulect), daclizumab (Zinbryta), and muromonab (Orthoclone).

56. The composition of claim 54 or 55, wherein the compound is selected from the group consisting of benzoyl peroxidase (BPO) and benzoyl peroxidase (OKT3).

59. The composition of any one of claims 54 to 58, wherein the poxvirus is a vaccinia virus.

60. 60. The composition of claim 59, wherein the vaccinia virus is selected from Dryvax, ACAM1000, ACAM2000, Lister, EM63, LIVP, Tian Tan, Copenhagen, Western Reserve, Modified Vaccinia Ankara (MVA), New York City Board of Health, Dalian, Ikeda, LC16M8, Western Reserve Copenhagen, Tashkent, Tian Tan, Wyeth, IHD-J, and IHD-W, Brighton, Dalian I, and Connaught strains.

61. 61. The composition of claim 60, wherein the vaccinia virus is ACAM1000 or ACAM2000.

62. 61. The composition of claim 60, wherein the vaccinia virus is a New York City Board of Health strain.

63. The composition according to any one of claims 54 to 62, wherein the poxvirus is an attenuated virus.

64. The composition of any one of claims 54 to 63, wherein the stem cell comprises a recombinant polynucleotide, said recombinant polynucleotide encoding a second therapeutic molecule.

65. The second therapeutic molecule may be selected from the group consisting of abatacept (Orencia), adalimumab (Humira), anakinra (Kineret), certolizumab (Cimzia), etanercept (Enbrel), golimumab (Simponi), infliximab (Remicade), ixekizumab (Taltz), natalizumab (Tysabri), rituximab (Rituxan), secukinumab (Cosentyx), tocilizumab (Actemra), ustekinumab (Stellara), vedolizumab (Entyvio), basiliximab (Simulect), daclizumab (Zinbryta), and muromonab (Orthoclone).

65. The composition of claim 64, wherein said compound is selected from the group consisting of benzoyl peroxidase (BPO) and benzoyl peroxidase (OKT3).

66. Stem cells include adult stem cells, embryonic stem cells, fetal stem cells, mesenchymal stem cells, neural stem cells, totipotent stem cells, pluripotent stem cells, multipotent stem cells, oligopotent stem cells, unipotent stem cells, adipose stromal cells, 66. The composition of any one of claims 54 to 65, selected from endothelial stem cells, induced pluripotent stem cells, bone marrow stem cells, umbilical cord blood stem cells, adult peripheral blood stem cells, myoblast stem cells, juvenile stem cells, dermal fibroblast stem cells, and combinations thereof.

67. The composition of any one of claims 54 to 66, wherein the stem cells are modified stem cells.

68. 68. The composition of claim 67, wherein the modified stem cell expresses a heterologous protein.

69. The composition of any one of claims 54 to 68, wherein the stem cells are derived from the subject to be treated with the composition.

70. The composition of any one of claims 54 to 69, wherein the stem cells are derived from a human.

71. The composition of any one of claims 54 to 69, wherein the stem cells are derived from a livestock animal.

72. The composition of any one of claims 54 to 69, wherein the stem cells are derived from a pet.

73. 73. The composition of claim 72, wherein the stem cells are derived from a canine subject.

74. The composition of any one of claims 54 to 73, wherein the therapeutic molecule treats an inflammatory or infectious disease.

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