Cell Population Having Immunoregulatory Activity, Method for Isolation and Uses

Abstract

The present invention provides a population of connective tissue derived cells that respond to interferon-gamma (IFN-γ) by expressing indolamine-2,3-dioxygenase (IDO) for use in preventing, treating or ameliorating one or more symptoms associated with disorders in which modulation of a subject's immune system is beneficial, including, but not limited to, autoimmune diseases, inflammatory disorders, and immunologically mediated diseases including rejection of transplanted organs and tissues.

1 . An isolated cell population comprising a cell population derived from connective tissue of a subject, wherein said cell population is characterised in that said cells:
 a) do not express markers specific for antigen-presenting cells (APC),   b) do not express indolamine 2,3-dioxygenase (IDO) constitutively,   c) express IDO upon stimulation with interferon-gamma (IFN-γ and,   d) present capacity to be differentiated into at least two cell lineages.   
     
     
         2 . Cell population according to  claim 1 , characterised in that it does not present tumorigenic activity. 
     
     
         3 . Cell population according to  claim 1 , characterised in that said cell population is negative for cell surface markers selected from the group consisting of CD11b, CD11c, CD14, CD31, CD34, CD45, CD133 and HLAII. 
     
     
         4 . Cell population according to  claim 1 , characterised in that said cell population is positive for at least one cell surface marker selected from the group consisting of CD9, CD44, CD54, CD90 and CD105. 
     
     
         5 . Cell population according to  claim 1 , characterised in that it is capable of being expanded ex vivo. 
     
     
         6 . Cell population according to  claim 1 , characterised in that said cell population it is isolated from tissues selected from the group consisting of adipose tissue, cartilaginous tissue, skin and bone marrow. 
     
     
         7 . Cell population according to  claim 1 , characterised in that said cell population is from human origin. 
     
     
         8 . Cell population according to  claim 1 , characterized in that it expresses at least one antigenic polypeptide. 
     
     
         9 . A method for isolating a cell population from connective tissue of a subject, wherein the cells of said cell population present a phenotype characterized in that (i) they do not express markers specific from APCs; (ii) they do not express IDO constitutively; (iii) they express IDO upon stimulation with IFN-γ, and (iv) they present capacity to be differentiated into at least two cell lineages, said method comprising:
 (i) preparing a cell suspension from a sample of a connective tissue from said subject;   (ii) recovering the cells from said cell suspension;   (iii) incubating said cells in a suitable cell culture medium on a solid surface under conditions which allow cells to adhere to the solid surface and proliferate;   (iv) washing said solid surface after incubation to remove non-adhered cells;   (v) selecting the cells which after being passaged at least twice in such medium remain adhered to said solid surface; and   (vi) confirming that the selected cell population presents the phenotype of interest.   
     
     
         10 . (canceled) 
     
     
         11 . Cell population according to  claim 1  for inducing transplantation tolerance, treating autoimmune diseases, or treating an inflammatory disease in a subject comprising administering to said subject an effective amount of a cell population of  claim 1 . 
     
     
         12 .- 14 . (canceled) 
     
     
         15 . Cell population according to  claim 11 , wherein said inflammatory disease is selected from the group consisting of Inflammatory Bowel Disease (IBD) and Rheumatoid Arthritis (RA). 
     
     
         16 .- 22 . (canceled) 
     
     
         23 . An isolated T-reg cell population from a subject, said T-reg cell population prepared or generated by the method comprising:
 (a) contacting a cell population according to  claim 1  with peripheral blood leukocytes, and   (b) selecting the T-reg cell population.   
     
     
         24 .- 25 . (canceled) 
     
     
         26 . A method for inducing transplantation tolerance, treating autoimmune diseases, or treating an inflammatory disease in a subject comprising administering to said subject an effective amount of a cell population of  claim 23 . 
     
     
         27 .- 29 . (canceled) 
     
     
         30 . The method according to  claim 26 , wherein said inflammatory disease is selected from the group consisting of Inflammatory Bowel Disease (IBD) and Rheumatoid Arthritis (RA). 
     
     
         31 .- 36 . (canceled) 
     
     
         37 . The method according to  claim 9  further comprising irradiating said cell population with a controlled source of ionizing radiation for an exposure time, wherein said exposure time is adjusted to impart a radiation dose that causes long term growth arrest of said cells. 
     
     
         38 . The isolated cell population according to  claim 1 , wherein said isolated cell population is irradiated with a controlled source of ionizing radiation for a exposure time, wherein said exposure time is adjusted to impart a radiation dose that causes long term growth arrest of said cell population. 
     
     
         39 . (canceled) 
     
     
         40 . A method for inducing transplantation tolerance, treating autoimmune diseases, or treating an inflammatory disease in a subject comprising administering to said subject an effective amount of a cell population of  claim 38 . 
     
     
         41 .- 43 . (canceled) 
     
     
         44 . The method according to  claim 40 , wherein said inflammatory disease is selected from the group consisting of Inflammatory Bowel Disease (IBD) and Rheumatoid Arthritis (RA). 
     
     
         45 .- 50 . (canceled) 
     
     
         51 . The method of  claim 9 , further comprising subjecting said cell population to treatment with Interferon-γ (IFN-γ). 
     
     
         52 . The isolated cell population according to  claim 1 , wherein said cell population is treated with IFN-γ. 
     
     
         53 . (canceled) 
     
     
         54 . A method for inducing transplantation tolerance, treating autoimmune diseases, or treating an inflammatory disease in a subject comprising administering to said subject an effective amount of a cell population of  claim 52 . 
     
     
         55 .- 57 . (canceled) 
     
     
         58 . The method of  claim 54 , wherein said inflammatory disease is selected from the group consisting of Inflammatory Bowel Disease (IBD) and Rheumatoid Arthritis (RA). 
     
     
         59 .- 64 . (canceled) 
     
     
         65 . The method according to  claim 9 , further comprising subjecting said cell population to (i) irradiation and (ii) stimulation with IFN-γ, wherein said treatments (i) and (ii) are carried out in any order. 
     
     
         66 . An isolated cell population according to  claim 1 , wherein said cell population is treated by (i) irradiation, and (ii) stimulation with IFN-γ, wherein treatments (i) and (ii) are carried out in any order. 
     
     
         67 . (canceled) 
     
     
         68 . A method for inducing transplantation tolerance, treating autoimmune diseases, or treating an inflammatory disease in a subject comprising administering to said subject an effective amount of a cell population of  claim 66 . 
     
     
         69 .- 71 . (canceled) 
     
     
         72 . The method of  claim 68 , wherein said inflammatory disease is selected from the group consisting of Inflammatory Bowel Disease (IBD) and Rheumatoid Arthritis (RA). 
     
     
         73 .- 78 . (canceled) 
     
     
         79 . A pharmaceutical composition comprising a cell population according to  claim 1  and an acceptable pharmaceutically carrier. 
     
     
         80 . A method for distinguishing adult multipotent cells from differentiated cells comprising
 (a) stimulating said adult multipotent stem cells with IFN-γ; and   verifying whether said adult multipotent stem cells express IDO upon stimulation with IFN-γ.   
     
     
         81 .- 84 . (canceled) 
     
     
         85 . A method for treating or ameliorating one or more symptoms associated with autoimmune diseases, inflammatory disorders, or immunologically mediated diseases, in a subject suffering from said disorders or diseases, which comprises administering to said subject in need of such treatment a prophylactically or therapeutically effective amount of a cell population according to  claim 1 . 
     
     
         86 . (canceled) 
     
     
         87 . Method according to  claim 85 , wherein said inflammatory disorder is selected from the group consisting of Inflammatory Bowel Disease (IBD) and Rheumatoid Arthritis (RA). 
     
     
         88 . An in vitro method of obtaining T-reg cells specific for a chosen antigen or group of antigens, which comprises:
 (a) contacting a cell population according to  claim 1  with said chosen antigen or group of antigens;   (b) bringing said cell population into contact with peripheral blood leukocytes; and   (c) selecting a T-reg cell population specific for said chosen antigen or group of antigens.   
     
     
         89 . A method of treating a disease or disorder related to a chosen antigen or group of antigens in a subject, said method comprising obtaining a T-reg cell population produced by the method of  claim 88  and administering said T-reg cells to the subject from which said peripheral blood leukocytes were obtained.