Transgenic mice having a human major histocompatibility complex (MHC) phenotype, experimental uses and applications

Abstract

The present invention-relates to transgenic mice and isolated transgenic mouse cells, the mice and mouse cells comprising a disrupted H2 class I gene, a disrupted H2 class II gene, a functional HLA class I transgene, and a functional HLA class II transgene. In embodiments, the transgenic mouse or mouse cells are deficient for both H2 class I and class II molecules, wherein the transgenic mouse comprises a functional HLA class I transgene and a functional HLA class II transgene. In embodiments, the transgenic mouse or mouse cell has the genotype HLA-A2 + HLA-DR1 + β2m°IAβ°. The invention also relates to methods of using a transgenic mouse of the invention.

1 . A transgenic mouse comprising:
 a) a disrupted H2 class I gene;   b) a disrupted H2 class II gene; and   c) a functional HLA class I or class II transgene.   
     
     
         2 . A transgenic mouse comprising:
 a) a disrupted H2 class I gene;   b) a disrupted H2 class II gene;   c) a functional HLA class I transgene; and   d) a functional HLA class II transgene.   
     
     
         3 . The transgenic mouse according to  claim 2 , wherein the HLA class I transgene is an HLA-A2 transgene and the HLA class II transgene is an HLA-DR1 transgene. 
     
     
         4 . The transgenic mouse according to  claim 3 , wherein the HLA-A2 transgene comprises the HLA-A2 sequence provided in the sequence listing and the HLA-DR1 transgene comprises the HLA-DR1 sequence provided in the sequence listing. 
     
     
         5 . A transgenic mouse deficient for both H2 class I and class II molecules, wherein the transgenic mouse comprises a functional HLA class I transgene and a functional HLA class II transgene. 
     
     
         6 . The transgenic mouse according to  claim 5 , having the genotype HLA-A2 + HLA-DR1 + β2m°IAβ°. 
     
     
         7 . The transgenic mouse according to  claim 6 , wherein the HLA-A2 transgene comprises the HLA-A2 sequence provided in the sequence listing and the HLA-DR1 transgene comprises the HLA-DR1 sequence provided in the sequence listing. 
     
     
         8 . A method of simultaneously identifying the presence of one or more epitopes in a candidate antigen or group of antigens, wherein the epitope elicits a specific humoral response, a TH HLA-DR1 restricted response, and/or a CTRL HLA-A2 restricted response, the method comprising:
 a) administering the candidate antigen or group of candidate antigens to the mouse of  claim 3  or  claim 6 ;   b) assaying for a specific humoral response in the mouse to the antigen;   c) assaying for a TH HLA-DR1 restricted response in the mouse to the antigen; and   d) assaying for a CTRL HLA-A2 restricted response in the mouse to the antigen; wherein,   observation of a specific humoral response in the mouse to the antigen identifies an epitope which elicits a humoral response in the antigen;   observation of a TH HLA-DR1 restricted response in the mouse to the antigen identifies an epitope which elicits a TH HLA-DR1 restricted response in the antigen; and   observation of a CTRL HLA-A2 restricted response in the mouse to the antigen identifies an epitope which elicits a CTRL HLA-A2 restricted response in the antigen.   
     
     
         9 . The method of  claim 8 , further comprising assaying for a Th1-specific response in the mouse to the antigen and assaying for a Th2-specific response in the mouse to the antigen; wherein
 observation of a Th1-specific response in the mouse to the antigen identifies an epitope which elicits a Th1-specific response in the mouse to the antigen; and   observation of a Th2-specific response in the mouse to the antigen identifies an epitope which elicits a Th2-specific response in the mouse to the antigen.   
     
     
         10 . A method of identifying the presence of an HLA DR1-restricted T helper epitope in a candidate antigen or group of candidate antigens, the method comprising:
 a) administering the candidate antigen or group of candidate antigens to the mouse of  claim 3  or  claim 6 ; and   b) assaying for a TH HLA-DR1 restricted T helper epitope response in the mouse to the antigen; wherein,   observation of a TH HLA-DR1 restricted T helper epitope response in the mouse to the antigen identifies an epitope which elicits a TH HLA-DR1 restricted T helper epitope response in the antigen.   
     
     
         11 . An isolated antigen comprising an HLA DR1-restricted T helper epitope identified by the method of  claim 10 . 
     
     
         12 . The isolated antigen of  claim 11 , wherein the antigen further comprises an epitope which elicits a humoral response and/or an epitope which elicits a CTRL HLA-A2 restricted response. 
     
     
         13 . The isolated antigen of  claim 11 , wherein the antigen comprising an HLA DR1-restricted T helper epitope comprises a polypeptide. 
     
     
         14 . The isolated antigen of  claim 11 , wherein the antigen comprising an HLA DR1-restricted T helper epitope comprises a polynucleotide. 
     
     
         15 . The isolated antigen of  claim 14 , wherein the antigen comprising an HLA DR1-restricted T helper epitope comprises, DNA, RNA, or DNA and RNA. 
     
     
         16 . A method of identifying the presence of an HLA-A2-restricted T cytotoxic (CTL) epitope in a candidate antigen or group of candidate antigens, the method comprising:
 a) administering the candidate antigen or group of candidate antigens to the mouse of  claim 3  or  claim 6 ; and   b) assaying for an HLA-A2-restricted T cytotoxic (CTL) response in the mouse to the antigen or group of antigens; wherein,   observation of an HLA-A2-restricted T cytotoxic (CTL) response in the mouse to the antigen or group of antigens identifies an epitope which elicits a an HLA-A2-restricted T cytotoxic (CTL) response in the antigen group of antigens.   
     
     
         17 . An isolated antigen comprising an HLA-A2-restricted T cytotoxic (CTL) epitope identified by the method of  claim 16 . 
     
     
         18 . The isolated antigen of  claim 17 , wherein the antigen further comprises an epitope which elicits a humoral response and/or an epitope which elicits a TH HLA-DR1 restricted T helper epitope response. 
     
     
         19 . The isolated antigen of  claim 17 , wherein the antigen comprising an HLA-A2-restricted T cytotoxic (CTL) epitope comprises a polypeptide. 
     
     
         20 . The isolated antigen of  claim 17 , wherein the antigen comprising an HLA-A2-restricted T cytotoxic (CTL) epitope comprises a polynucleotide. 
     
     
         21 . The isolated antigen of  claim 20 , wherein the antigen comprising an HLA-A2-restricted T cytotoxic (CTL) epitope comprises, DNA, RNA, or DNA and RNA. 
     
     
         22 . A method of comparing the efficiency of T-helper cell response induced by two or more vaccines, the method comprising:
 a) administering a first candidate vaccine to a mouse of  claim 3  or  claim 6  and measuring the T-helper cell response induced in the mouse by the first candidate vaccine;   b) administering a second candidate vaccine to a mouse of  claim 3  or  claim 6  and measuring the T-helper cell response induced in the mouse by the second candidate vaccine;   c) administering each additional candidate vaccine to be compared to a mouse of  claim 3  or  claim 6  and measuring the T-helper cell response induced in the mouse by each additional candidate vaccine to be compared; and   d) determining the efficiency of each candidate vaccine to induce a T-helper cell response by comparing the T-helper cell responses to each of the vaccines to be compared with each other.   
     
     
         23 . The method of  claim 22 , wherein the T-helper cell response is an HLA-DR1 restricted response. 
     
     
         24 . A method of comparing the efficiency of T cytotoxic cell response induced by two or more vaccines, the method comprising:
 a) administering a first candidate vaccine to a mouse of  claim 3  or  claim 6  and measuring the T cytotoxic cell response induced in the mouse by the first candidate vaccine;   b) administering a second candidate vaccine to a mouse of  claim 3  or  claim 6  and measuring the T cytotoxic cell response induced in the mouse by the second candidate vaccine;   c) administering each additional candidate vaccine to be compared to a mouse of  claim 3  or  claim 6  and measuring the T cytotoxic cell response induced in the mouse by each additional candidate vaccine to be compared; and   d) determining the efficiency of each candidate vaccine to induce a T cytotoxic cell response by comparing the T cytotoxic cell responses to each of the vaccines to be compared with each other.   
     
     
         25 . The method of  claim 24 , wherein the T cytotoxic cell response is an HLA-A2 restricted response. 
     
     
         26 . A method of simultaneously comparing the efficiency of T-helper cell response and T cytotoxic cell response induced by two or more vaccines, the method comprising:
 a) administering a first candidate vaccine to a mouse of  claim 3  or  claim 6  and measuring the T-helper cell response and T cytotoxic cell response induced in the mouse by the first candidate vaccine;   b) administering a second candidate vaccine to a mouse of  claim 3  or  claim 6  and measuring the T-helper cell response and T cytotoxic cell response induced in the mouse by the second candidate vaccine;   c) administering each additional candidate vaccine to be compared to a mouse of  claim 3  or  claim 6  and measuring the T-helper cell response and T cytotoxic cell response induced in the mouse by each additional candidate vaccine to be compared; and   d) determining the efficiency of each candidate vaccine to induce a T-helper cell response and T cytotoxic cell response by comparing the T-helper cell response and T cytotoxic cell response to each of the vaccines to be compared with each other.   
     
     
         27 . The method of  claim 26 , wherein the T-helper cell response is an HLA-DR1 restricted response, and wherein the T cytotoxic cell response is an HLA-A2 restricted response. 
     
     
         28 . A method of simultaneously determining the humoral response, the T-helper cell response, and the T cytotoxic cell response of a mouse following its immunization with an antigen or a vaccine comprising one or more antigens, the method comprising:
 a) administering the antigen or the vaccine comprising one or more antigens to a mouse of  claim 3  or  claim 6 ;   b) assaying for a specific humoral response in the mouse to the antigen or vaccine comprising one or more antigens;   c) assaying for a T-helper cell response in the mouse to the antigen or vaccine comprising one or more antigens; and   d) assaying for a T cytotoxic cell response in the mouse to the antigen or vaccine comprising one or more antigens.   
     
     
         29 . The method of  claim 28 , wherein the T-helper cell response is a TH HLA-DR1 restricted response. 
     
     
         30 . The method of  claim 28 , wherein the T cytotoxic cell response is a CTRL HLA-A2 restricted response. 
     
     
         31 . A method of optimizing two or more candidate vaccine compositions for administration to a human, based on preselected criteria, the method comprising:
 simultaneously determining the humoral response, the T-helper cell response, and the T cytotoxic cell response of a mouse following its immunization with the two or more candidate vaccine compositions, according to  claim 28 ; and   selecting an optimized vaccine by applying preselected criteria to the results.   
     
     
         32 . The method according to  claim 31 , wherein the two or more candidate vaccines differ only in the ratio of antigen to adjuvant present in the vaccine. 
     
     
         33 . The method according to  claim 31 , wherein the two or more candidate vaccines differ only in the type of adjuvant present in the vaccine. 
     
     
         34 . A method of determining whether a vaccine poses a risk of induction of an autoimmune disease when administered to a human, the method comprising:
 a) administering the vaccine to a mouse of  claim 3  or  claim 6 ; and   b) assaying for an autoimmune response in the mouse; wherein, observation of an autoimmune response in the mouse indicates that the vaccine poses a risk of induction of an autoimmune disease when administered to a human.   
     
     
         35 . An isolated transgenic mouse cell comprising:
 a) a disrupted H2 class I gene;   b) a disrupted H2 class II gene; and   c) a functional HLA class I or class II transgene.   
     
     
         36 . An isolated transgenic mouse cell comprising:
 a) a disrupted H2 class I gene;   b) a disrupted H2 class II gene;   c) a functional HLA class I transgene; and   d) a functional HLA class II transgene.   
     
     
         37 . The transgenic mouse cell according to  claim 36 , wherein the HLA class I transgene is an HLA-A2 transgene and the HLA class II transgene is an HLA-DR1 transgene. 
     
     
         38 . The transgenic mouse cell according to  claim 37 , wherein the HLA-A2 transgene comprises the HLA-A2 sequence provided in the sequence listing and the HLA-DR1 transgene comprises the HLA-DR1 sequence provided in the sequence listing. 
     
     
         39 . An isolated transgenic mouse cell deficient for both H2 class I and class II molecules, wherein the transgenic mouse cell comprises a functional HLA class I transgene and a functional HLA class II transgene. 
     
     
         40 . The transgenic mouse cell according to  claim 39 , having the genotype HLA-A2 + HLA-DR1 + β2m°IAβ°. 
     
     
         41 . The transgenic mouse cell according to  claim 40 , wherein the HLA-A2 transgene comprises the HLA-A2 sequence provided in the sequence listing and the HLA-DR1 transgene comprises the HLA-DR1 sequence provided in the sequence listing.