Ex-vivo priming for generating cytotoxic T lymphocytes specific for non-tumor antigens to treat autoimmune and allergic disease

Abstract

Cytotoxic T lymphocytes (CTLs) specific for antigenic peptides derived from IgE molecule can be generated in vitro by stimulating resting naive CD8 T cells with IgE peptides presented by artificial antigen presenting cells. The IgE specific CTLs lyse the target cells loaded with IgE peptides in vitro and inhibit antigen specific IgE response in vivo. In addition, adoptive transfer of the IgE specific CTL to an asthmatic mouse model can inhibit the development of lung inflammation and airway hypersensitivity. IgE specific CTL provides a treatment for allergic asthma and other IgE-mediated allergic diseases. Antigenic peptides identified from non-tumor self-antigens induce specific cytotoxic T lymphocyte (CTL) in vitro. The CTL induced by peptides identified from CD40L can kill activated CD4 T cells. In vitro generated CTL specific for CD40L inhibit CD4-dependent antibody responses of all isotypes in vivo. In contrast, CTL induced by antigenic peptides derived from IgE specifically inhibit IgE responses, and adoptive transfer of CD40L-specific CTL to NOD mice at early age delay the development of diabetes in NOD mice. In vitro generated CTL specific for non-tumor self-antigens expressed on activated CD4 T cells regulate immune responses in vivo.

1 . A method for producing CTL specific for one or more non-tumor self antigen T cell epitopes, comprising: 
 a) isolating CD8+ T cells from a subject;    b) loading antigen presenting cells (APCs) having Class I MHC molecules with the non-tumor self antigen T cell epitopes,    c) culturing the CD8+ T cells with the APCs for a period of time sufficient for activation of precursor CD8+ T cells specific for the T cell epitopes;    d) expanding in culture the activated CD8+ T cells in the presence of components required for proliferation of the activated CD8+ T cells; and    e) collecting CD8+ T cells from the culture.    
     
     
         2 . The method of  claim 1  wherein the T cell epitopes are present on IgE protein.  
     
     
         3 . The method of  claim 1  wherein the T cell epitopes are present on CD40 ligand protein.  
     
     
         4 - 6 . (canceled)  
     
     
         7 . A method for treating a disease mediated by a disease causing target cell, wherein the target cell has on its surface one or more non-tumor self antigen T cell epitopes associated with Class I MHC molecules, comprising administering to a patient in need of such treatment, activated CD8+ T cells wherein the CD8+ T cells have been selectively activated by culturing said CD8+ T cells in the presence of antigen presenting cells (APC) having Class I MHC molecules and wherein said APC are loaded with one of said non-tumor self antigen T cell epitopes.  
     
     
         8 . The method of  claim 7  wherein the T cell epitopes are present on IgE protein.  
     
     
         9 . The method of  claim 7  wherein the T cell epitopes are present on CD40 ligand protein.  
     
     
         10 . The method of  claim 2  wherein said non tumor self-antigen T cell epitopes are epitopes present on IgE protein and are selected from the group consisting of TQSPSVFPL (SEQ ID NO:64), SLNGTTMTL (SEQ ID NO:50), TMTLPATTL (SEQ ID NO:69), TLPATTLTL (SEQ ID NO:67), TLSGHYATI (SEQ ID NO:68), WVDNKTFSV (SEQ ID NO:51), WLSDRTYTC (SEQ ID NO:52), ALMRSTTKT (SEQ ID NO:65), NFMPEDISV (SEQ ID NO:79), YATISLLTV (SEQ ID NO:58), TLTVTSTLPV (SEQ ID NO:56), SVQWLHNEV (SEQ ID NO:76), QWLHNEVQL (SEQ ID NO:80), TLACLIQNFM (SEQ ID NO:59), or QVMDVDLSTA (SEQ ID NO:60).  
     
     
         11 . The method of  claim 8  wherein said non tumor self-antigen T cell epitopes are epitopes present on IgE protein and are selected from the group consisting of TQSPSVFPL (SEQ ID NO:64), SLNGTTMTL (SEQ ID NO:50), TMTLPATTL (SEQ ID NO:69), TLPATTLTL (SEQ ID NO:67), TLSGHYATI (SEQ ID NO:68), WVDNKTFSV (SEQ ID NO:51), WLSDRTYTC (SEQ ID NO:52), ALMRSTTKT (SEQ ID NO:65), NFMPEDISV (SEQ ID NO:79), YATISLLTV (SEQ ID NO:58), TLTVTSTLPV (SEQ ID NO:56), SVQWLHNEV (SEQ ID NO:76), QWLHNEVQL (SEQ ID NO:80), TLACLIQNFM (SEQ ID NO:59), or QVMDVDLSTA (SEQ ID NO:60).  
     
     
         12 . The method of  claim 8  wherein the disease is an allergic disease, allergic systemic anaphylaxis, allergic rhinitis, hay fever, food allergy, or allergic asthma.  
     
     
         13 . The method of  claim 9  wherein the disease is autoimmune disease.  
     
     
         14 . The method of  claim 13  wherein the disease is insulin dependent diabetes, rheumatoid arthritis, SLE, multiple sclerosis, psoriasis, autoimmune nephritis, multiple sclerosis, autoimmune thyroiditis, Crohn's disease, inflammatory bowel disease, graft versus host disease, or transplant rejection.  
     
     
         15 . The method of  claim 9  wherein the disease is an inflammatory disease.