Use of apoptotic cells ex vivo to generate regulatory t cells

Abstract

Many cell types in the body can remove apoptotic and cellular debris from tissues; however, the professional phagocyte, or antigen presenting cell (“APC”), has a high capacity to do so. The recognition of apoptotic cells (“ACs”) occurs via a series of evolutionarily-conserved, AC associated molecular-pattern receptors (“ACAMPRs”) on APCs that recognize and bind corresponding apoptotic-cell-associated molecular patterns (“ACAMPs”). These receptors recognize ligands such as phosphotidyl serine and oxidized lipids found on apoptotic cells. Savill et al. (2002); and Gregory et al. (2004).

1 - 17 . (canceled) 
     
     
         18 . A composition comprising a population of T cells with regulatory activity (T regs) obtained by incubating leukocytes with autologous apoptotic peripheral blood mononuclear cells (ACs) and at least one factor that further enhances generation or function of the T regs, wherein the factor is selected from the group consisting of hormones, proteins, drugs or antibodies. 
     
     
         19 . The composition according to  claim 18  further comprising the step of selecting leukocytes expressing CD4 to obtain CD4+ cells. 
     
     
         20 . The composition according to  claim 19  wherein the incubation is at about a 1:10 to about a 10:1 ratio CD4+:ACs. 
     
     
         21 . The composition according to  claim 20  wherein the incubation is at about a 2:1 to about a 1:2 ratio CD4+:ACs. 
     
     
         22 . The composition according to  claim 18  wherein the ACs are obtained by an apoptosis-inducing treatment. 
     
     
         23 . The composition according to  claim 22  wherein the apoptosis-inducing treatment is an ECP procedure that employs a photoactivable compound together with light of a wavelength that activates the photoactivable compound. 
     
     
         24 . The composition according to  claim 18  wherein the factor is IL-IO. 
     
     
         25 . The composition according to  claim 24  wherein the IL-10 is present at a concentration of about 1 ng/ml to about 100 ng/ml. 
     
     
         26 . The composition according to  claim 18  wherein the incubation is for about 1 to about 14 days. 
     
     
         27 . The composition according to  claim 18  further comprising adding an antigen to the incubation to generate Tregs which regulate immune response to the antigen. 
     
     
         28 . The composition according to  claim 27  wherein the antigen is an alloantigen. 
     
     
         29 . A method of treating autoimmune disorder or ameliorating one or more symptoms thereof, comprising administering to a patient in need thereof an effective amount of a composition comprising a population of T cells with regulatory activity (T regs) obtained by incubating autologous leukocytes with autologous apoptotic peripheral blood mononuclear cells (ACs). 
     
     
         30 . The method according to  claim 29  wherein the apoptosis-inducing treatment is an ECP procedure that employs a photoactivable compound together with light of a wavelength that activates the photoactivable compound. 
     
     
         31 . The method according to  claim 29  wherein the incubation further comprises adding one or more factors that further enhance generation or function of the T regs, wherein the factor is selected from the group consisting of hormones, proteins, drugs or antibodies. 
     
     
         32 . The method according to  claim 29  wherein the factors are selected from at least one of TGFβ, αMSH, anti-CD46, IL-IO, vitamin D 3 , dexamethasone, rapamycin and IL-2. 46. The method according to claim  45  wherein the factor is IL-IO. 
     
     
         33 . The method according to  claim 29  wherein the IL-10 is present at a concentration of about 1 ng/ml to about 100 ng/ml. 
     
     
         34 . The method according to  claim 29  further comprising adding an antigen to the incubation to generate Tregs which regulate immune response to the antigen. 
     
     
         35 . The method according to  claim 34  wherein the antigen is an alloantigen. 
     
     
         36 . The method according to  claim 29  wherein the autoimmune disorder is at least one of acute transverse myelitis, alopecia areata, Alzheimer's disease, amyotrophic lateral sclerosis, ankylosing spondylitis, antiphospholipid syndrome, atherosclerosis, autoimmune Addison's disease, autoimmune hemolytic anemia, Behcet's disease, bullous pemphigoid, cardiomyopathy, celiac sprue-dermatitis, Cerebellar Spinocerebellar Disorders, spinocerebellar degenerations (spinal ataxia, Friedreich's ataxia, cerebellar cortical degenerations), chronic alcoholism, alcohol-induced hepatitis, autoimmune hepatitis, chronic fatigue immune dysfunction syndrome (CFIDS), chronic inflammatory bowel disease, chronic inflammatory demyelinating polyneuropathy, Churg-Strauss syndrome, cicatricial pemphigoid, cold agglutinin disease, CResT syndrome, Creutzfeldt-Jakob disease, Crohn's disease, Dejerine-Thomas atrophy, Dementia pugilistica, diabetes mellitus, Diffuse Lewy body disease, discoid lupus, disorders of the basal ganglia, disseminated intravascular coagulation, Down's Syndrome in middle age, drug-induced movement disorders, essential mixed cryoglobulinemia, fibromyalgia-fibromyositis, graft versus host disease, Graves' disease, <3uillain-Barre syndrome, Hallerrorden-Spatz disease, Hashimoto's thyroiditis, Huntington's Chorea senile chorea, idiopathic pulmonary fibrosis, idiopathic thrombocytopenia purpura, (ITP), IgA nephropathy, infantile or juvenile spinal muscular atrophy, insulin dependent diabetes, juvenile arthritis, Kawasaki's pathology, lesions of the corticospinal system, Leukemias, Hodgkin's lymphoma, non-Hodgkin's lymphoma, lichen planus, Meniere's disease, mixed connective tissue disease, multiple sclerosis, multiple systems degenerations (Mencel, Dejerine-Thomas, Shy-Drager,
 Machado-Joseph), myasthenia gravis, neurogenic muscular, Parkinson's disease, pemphigus vulgaris, pernicious anemia, polyarteritis nodosa, polychondritis, polyglandular syndromes, polymyalgia rheumatica, polymyositis dermatomyositis, primary agammaglobulinemia, primary biliary cirrhosis, Progressive supranuclear palsy, psoriasis, Raynaud's phenomenon, Reiter's syndrome, rheumatic fever, rheumatoid arthritis, sarcoidosis, scleroderma, Senile Dementia of Lewy body type, Sjogren's syndrome, stiff-man syndrome, Subacute sclerosing 
 panencephalitis, systemic disorders (Refsum's disease, abetalipoprotemia, ataxia, telangiectasia, mitochondrial multi-system disorder), systemic lupus erythematosus (SLE), Takayasu arteritis, temporal arteritis/giant cell arteritis, thyroidosis, ulcerative colitis, uveitis, vasculitis, vitiligo, Wegener's granulomatosis, Wernicke-Korsakoff syndrome. 
 
     
     
         37 . A method of treating patient with a disorder or the predisposition for a disorder comprising testing the patient to determine whether the patient has a disorder, and administering to a patient in need thereof an effective amount of a composition comprising a population of T cells with regulatory activity (T regs) obtained by incubating autologous leukocytes with autologous apoptotic peripheral blood mononuclear cells (ACs).