Beta-2 microglobulin-deficient cells

Abstract

The invention provides isolated primate cells preferably human cells that comprise a genetically engineered disruption in a beta-2 microglobulin (B2M) gene, which results in deficiency in MHC class I expression and function. Also provided are the method of using the cells for transplantation and treating a disease condition.

We claim: 
     
         1 . An isolated cell comprising a genetically engineered disruption in a beta-2 microglobulin (B2M) gene, wherein the cell is a primate cell. 
     
     
         2 . The cell of  claim 1 , wherein the cell comprises genetically engineered disruptions of all copies of the B2M gene. 
     
     
         3 . The cell of  claim 1 , wherein the cell is a human cell. 
     
     
         4 . The cell of any  claim 1 , wherein the cell further comprises one or more recombinant immunomodulatory genes, each capable of expressing an immunomodulatory polypeptide in the cell. 
     
     
         5 . The cell of  claim 4 , wherein the one or more immunomodulatory genes comprise a polynucleotide capable of encoding a single chain fusion human leukocyte antigen (HLA) class I protein. 
     
     
         6 . The cell of  claim 5 , wherein the single chain fusion HLA class I protein comprises at least a portion of B2M covalently linked to at least a portion of an HLA class I α chain selected from the group consisting of HLA-A, HLA-B, HLA-C, HLA-E, HLA-F and HLA-G. 
     
     
         7 . The cell of  claim 5 , wherein the single chain fusion HLA class I protein comprises at least a portion of B2M and at least a portion of HLA class I α chain is selected from the group consisting of HLA-E, HLA-G and HLA-A. 
     
     
         8 .- 14 . (canceled) 
     
     
         15 . The cell of  claim 5 , wherein the cell further expresses a target peptide antigen that is presented by the single chain fusion HLA class I protein on the cell surface. 
     
     
         16 . The cell of  claim 15  wherein the target peptide antigen is covalently linked to the single chain fusion HLA class I protein. 
     
     
         17 . The cell of  claim 1 , wherein the cell further comprises one or more recombinant genes capable of encoding a suicide gene product. 
     
     
         18 . The cell of  claim 17  wherein the suicide gene product comprises a protein selected from the group consisting of thymidine kinase and an apoptotic signaling protein. 
     
     
         19 . The cell of any  claim 1 , wherein the cell has a normal karyotype. 
     
     
         20 . The cell of  claim 1 , wherein the cell is a non-transformed cell. 
     
     
         21 . The cell of  claim 1 , wherein the cell is a stem cell. 
     
     
         22 .- 25 . (canceled) 
     
     
         26 . A vaccine comprising the cell of  claim 15 , wherein the vaccine is capable of eliciting in a primate an immune response specific for the target peptide antigen. 
     
     
         27 . The vaccine of  claim 26 , wherein the primate is a human and the cell is a human cell. 
     
     
         28 . A method of transplantation in a patient in need thereof comprising the step of administering to the patient an effective amount of the cell of  claim 1 . 
     
     
         29 - 31 . (canceled) 
     
     
         32 . The method of  claim 28 , wherein the cell further expresses a single chain fusion HLA class I protein. 
     
     
         33 . A method of treating a disease condition in a patient in need thereof comprising the step of administering to the patient an effective amount of the cell of  claim 1 , wherein the disease condition is diabetes, an autoimmune disease, cancer, infection, anemia, cytopenia, myocardial infarction, heart failure, skeletal or joint condition, osteogenesis imperfecta or burns. 
     
     
         34 .- 38 . (canceled) 
     
     
         39 . A kit comprising the cell of  claim 1 . 
     
     
         40 .- 43 . (canceled)