Cd4+ cd25+ t-cells activated to a specific antigen

Abstract

The invention relates to a method of assessing whether a subject includes CD4+,CD25+ T cells that have been activated to a specific antigen. The method includes the steps of obtaining from the subject a sample of lymphocytes including CD4+,CD25+ T cells, incubating at least one portion of the sample of lymphocytes so as to promote, distinction of CD4+,CD25+ T cells that have been activated to the specific antigen from those CD4+,CD25+ T cells that have not been activated to the specific antigen, and thereafter determining whether CD4+,CD25+ T cells activated to the specific antigen are present in the sample. The invention further relates to methods of growing CD4+, CD25+ T cells that have been activated to a specific antigen in vitro and to methods of increasing tolerance in a subject using the CD4+, CD25+ T cells that have been grown in vitro.

1 - 65 . (canceled) 
     
     
         66 . A method of increasing tolerance to a specific antigen in a subject in need thereof, comprising administering to the subject an effective amount of CD4 + ,CD25 +  T cells activated to the specific antigen grown in vitro by culturing CD4 + ,CD25 +  T cells activated to the specific antigen in the presence of at least one cytokine selected from the group consisting of IL-5, IL-12, IL-23 and IFN-γ or a biologically active fragment thereof, or a functionally equivalent molecule thereof. 
     
     
         67 . The method of  claim 66 , wherein the CD4 + ,CD25 +  T cells activated to the specific antigen are contacted with the specific antigen in vitro prior to, or simultaneous with, culturing the activated T cells in the presence of the at least one cytokine. 
     
     
         68 . The method of  claim 66 , further comprising administering to the subject one or more antibodies which specifically bind to CD4 + ,CD25 −  T cells. 
     
     
         69 . The method of  claim 66 , wherein the CD4 + ,CD25 +  T cell activated to the specific antigen are activated to the specific antigen in vitro by contacting naïve CD4 + ,CD25 +  T cells with the specific antigen in vitro, and culturing the CD4 + ,CD25 +  T cells in the presence of one or more cytokines capable of supporting activation of CD4 + ,CD25 +  T cells. 
     
     
         70 . The method of  claim 69 , wherein the one or more cytokines capable of supporting activation of CD4 + ,CD25 +  T cells are selected from the group consisting of IL-2, a biologically active fragment thereof, or a functionally equivalent molecule thereof, and IL-4, a biologically active fragment thereof, or a functionally equivalent molecule thereof. 
     
     
         71 . The method of  claim 66 , wherein the CD4 + ,CD25 +  T cells activated to the specific antigen are activated to the specific antigen in vivo prior to culturing in vitro. 
     
     
         72 . The method of  claim 66 , wherein a mixture of naïve and activated CD4 + ,CD25 +  T cells are grown by contacting naïve and activated CD4 + ,CD25 +  T cells with the specific antigen in vitro, and culturing the naïve and activated CD4 + ,CD25 +  T cells in the presence of IL-2, a biologically active fragment thereof, or a functionally equivalent molecule thereof, and/or IL-4, a biologically active fragment thereof, or a functionally equivalent molecule thereof, and at least one cytokine selected from the group consisting of IL-5, IL-12, IL-23 and IFN-γ, or a biologically active fragment thereof, or a functionally equivalent molecule thereof. 
     
     
         73 . The method of  claim 66 , wherein the at least one cytokine has been added exogenously to medium in which the CD4 + ,CD25 +  T cells are cultured. 
     
     
         74 . The method of  claim 66 , further comprising administering to the subject an effective amount of at least one cytokine selected from the group consisting of IL-5, IL-12, IL-23 and IFN-γ or a biologically active fragment thereof, or a functionally equivalent molecule thereof, simultaneously with, or subsequent to, administering the activated CD4 + ,CD25 +  T cells. 
     
     
         75 . The method of  claim 66 , wherein the specific antigen is autoantigen, an alloantigen, a xenoantigen, a tumour antigen, an allergen, or an antigen from an infectious agent. 
     
     
         76 . A method for treating or preventing in a subject in need thereof a disease resulting from an immune response to a specific antigen, the method comprising administering to the subject a therapeutically effective amount of CD4 + ,CD25 +  T cells activated to the specific antigen grown in vitro by culturing CD4 + ,CD25 +  T cells activated to the specific antigen in vitro in the presence of at least one cytokine selected from the group consisting of IL-5, IL-12, IL-23 and IFN-γ, or a biologically active fragment thereof, or a functionally equivalent molecule thereof. 
     
     
         77 . The method of  claim 76 , wherein the CD4 + ,CD25 +  T cells activated to the specific antigen are contacted with the specific antigen in vitro prior to, or simultaneous with, culturing the activated CD4 + ,CD25 +  T cells in the presence of the at least one cytokine. 
     
     
         78 . The method of  claim 76 , wherein the disease is: (a) a disease associated with an immune response to an autoantigen; (b) a disease that is the result of an immune response to a non-self antigen in contact with the subject; or (c) a disease that is the result of an immune response to an allergen in contact with the subject. 
     
     
         79 . A method of inducing tolerance to a specific antigen in a subject in need thereof, wherein the subject's immune system is exposed to the specific antigen, the method comprising administering to the subject an effective amount of at least one cytokine selected from the group consisting of IL-5, IL-12, IL-23 and IFN-γ, or a biologically active fragment thereof, or a functionally equivalent molecule thereof. 
     
     
         80 . A method of treating or preventing in a subject in need thereof a disease resulting from an immune response to an antigen, the method comprising administering to the subject an effective amount of at least one cytokine selected from the group consisting of IL-5, IL-12, IL-23 and IFN-γ, or a biologically active fragment thereof, or a functionally equivalent molecule thereof. 
     
     
         81 . The method of  claim 80 , wherein the disease is: (a) a disease associated with an immune response to an autoantigen; (b) a disease that is the result of an immune response to a non-self antigen in contact with the subject; or (c) a disease that is the result of an immune response to an allergen in contact with the subject. 
     
     
         82 . A method of inducing tolerance to a specific antigen in a subject in need thereof, wherein the subject's immune system is exposed to the specific antigen, the method comprising:
 (a) determining whether a sample of lymphocytes comprising CD4 + ,CD25 +  T cells obtained from a subject comprises CD4+,CD25+T cells that are responsive to one or more cytokines selected from the group consisting of IL-5, IL-12, IL-23 and IFN-γ, or a biologically active fragment thereof, or a functionally equivalent molecule thereof; and   (b) administering an effective amount of one or more cytokines selected from the group consisting of IL-5, IL-12, IL-23 and IFN-γ, or a biologically active fragment thereof, or a functionally equivalent molecule thereof, to which the CD4 + ,CD25 +  T cells are responsive.   
     
     
         83 . The method of  claim 82  comprising, prior to or simultaneously with, step (b), the step of administering to the subject an effective amount of one or more cytokines selected from the group consisting of IL-2, a biologically active fragment thereof, or a functionally equivalent molecule thereof, and IL-4, a biologically active fragment thereof, or a functionally equivalent molecule thereof. 
     
     
         84 . A method of treating or preventing in a subject in need thereof a disease resulting from an immune response to an antigen, the method comprising:
 (a) determining whether a sample of lymphocytes comprising CD4+,CD25+T cells obtained from a subject comprises CD4 + ,CD25 +  T cells that are responsive to one or more cytokines selected from the group consisting of IL-5, IL-12, IL-23 and IFN-γ, or a biologically active fragment thereof, or a functionally equivalent molecule thereof; and   (b) administering an effective amount of one or more cytokines selected from the group consisting of IL-5, IL-12, IL-23 and IFN-γ, or a biologically active fragment thereof, or a functionally equivalent molecule thereof, to which the CD4 + ,CD25 +  T cells are responsive.   
     
     
         85 . The method of  claim 84  comprising, prior to or simultaneously with, step (b), the step of administering to the subject an effective amount of one or more cytokines selected from the group consisting of IL-2, a biologically active fragment thereof, or a functionally equivalent molecule thereof, and IL-4, a biologically active fragment thereof, or a functionally equivalent molecule thereof. 
     
     
         86 . The method of  claim 84 , wherein the disease is: (a) a disease associated with an immune response to an autoantigen; (b) a disease that is the result of an immune response to a non-self antigen in contact with the subject; or (c) a disease that is the result of an immune response to an allergen in contact with the subject.