Materials and methods relating to improved vaccinations strategies

Abstract

The invention provides novel vaccination strategies based on a prime-boost vaccination regimen. The inventors have determined improved ways of boosting an immune response in a patient previously primed or exposed to a plurality of epitopes. The improved method requires the epitopes in the boosting phase to be administered individually, i.e. held on separate peptide constructs.

1 . A method of boosting an immune response to a plurality of epitopes in an individual, said individual having been previously exposed to at least one of said plurality of epitopes; said method comprising the steps of administering to the individual a composition comprising a plurality of constructs each construct comprising one of said plurality of eptiopes.  
     
     
         2 . A method according to  claim 1  wherein said constructs are nucleic acid sequences each encoding one of said plurality of epitopes.  
     
     
         3 . A method according to  claim 1  wherein said constructs are selected from the group consisting of peptide, polypeptide and protein.  
     
     
         4 . A method according to  claim 1  wherein said composition comprises a mixture of one or more nucleic acid constructs each encoding one of said plurality of epitopes and one or more peptide constructs each comprising one of said plurality of epitopes.  
     
     
         5 . A method according to  claim 2  further comprising a vehicle for carrying said construct.  
     
     
         6 . A method according to  claim 5  wherein said vehicle is a nucleic acid expression vector.  
     
     
         7 . A method according to  claim 6  wherein said nucleic acid expression vector is a viral vector.  
     
     
         8 . A method according to  claim 7  wherein said viral vector is selected from the group consisting of adenovirus vector, Herpes simplex virus vector, vaccinia virus vector, avipox virus vector and alphavirus vector.  
     
     
         9 . A method according to  claim 8  wherein said alphavirus vector is semliki forest virus vector.  
     
     
         10 . A method according to  claim 3  further comprising a vehicle for carrying said construct.  
     
     
         11 . A method according to  claim 10  wherein said vehicle is a cell.  
     
     
         12 . A method according to  claim 11  wherein said cell is an antigen presenting cell.  
     
     
         13 . A method according to  claim 12  wherein said antigen presenting cell is a dendritic cell or a tumour cell.  
     
     
         14 . A method according to  claim 12  wherein said antigen presenting cell is a lymphocyte.  
     
     
         15 . A method according to  claim 1  wherein the immune response is a CD8 +  T cell immune response.  
     
     
         16 . A method according to  claim 1  wherein said plurality of epitopes are from a self antigen.  
     
     
         17 . A method according to  claim 1  wherein said plurality of epitopes are from a tumour antigen.  
     
     
         18 . A method according to  claim 17  wherein the tumour is melanoma.  
     
     
         19 . A method according to  claim 1  wherein said plurality of epitopes are from a pathogen.  
     
     
         20 . A method according to  claim 19  wherein said pathogen is influenza virus.  
     
     
         21 . A method according to  claim 1  wherein said individual is a test animal and the epitopes are test epitopes.  
     
     
         22 . A method according to  claim 21  further comprising the step of-determining the CTL response to each of the epitopes under test.  
     
     
         23 . A method according to  claim 21  wherein the test animal is a transgenic animal that provides an immune enviroment close to the human immune environment.  
     
     
         24 . A method according to  claim 23  wherein the test animal is an A2 transgenic mouse.  
     
     
         25 . A method of inducing an immune response to a plurality of epitopes in an individual, said method comprising the steps of 
 a) administering to the individual a priming composition comprising one or more priming constructs comprising one or more of said plurality of epitopes; and    b) administering a boosting composition comprising a plurality of boosting constructs each comprising one of said plurality of epitopes; wherein said boosting composition is administered after the priming composition.    
     
     
         26 . A method according to  claim 25  wherein the one or more priming construct is a nucleic acid sequence encoding one or more of said plurality of epitopes.  
     
     
         27 . A method according to  claim 25  wherein said boosting constructs are nucleic acid sequences each encoding one of said plurality of epitopes.  
     
     
         28 . A method according to  claim 25  wherein the one or more priming construct is a peptide, polypeptide or protein comprising one or more of said plurality of epitopes.  
     
     
         29 . A method according to  claim 25  wherein said boosting constructs selected from the group consisting of peptide, polypeptide or protein.  
     
     
         30 . A method according to  claim 25  wherein said boosting composition comprises a mixture of one or more nucleic acid sequence constructs each encoding one of said plurality of epitopes and one or more peptide, polypeptide or protein constructs each comprising one of said plurality of epitopes.  
     
     
         31 . A method according to  claim 25  wherein said priming composition further comprises one or more vehicles for carrying said one or more constructs.  
     
     
         32 . A method according to  claim 25  wherein said boosting composition further comprises a plurality of vehicles for carrying said plurality of boosting constructs.  
     
     
         33 . A method according to  claim 31  wherein said one or more vehicles is selected from the group consisting of a nucleic acid expression vector and a cell.  
     
     
         34 . A method according to  claim 32  wherein said plurality of vehicles are selected from the group consisting of nucleic acid expression vector and a cell.  
     
     
         35 . A method according to  claim 33  or  claim 34  wherein said nucleic acid expression vector is a viral vector.  
     
     
         36 . A method according to  claim 35  wherein said viral vector is selected from the group consisting of adenovirus vector, Herpes simplex virus vector, vaccinia virus vector, avipox vector and alphavirus vector.  
     
     
         37 . A method according to  claim 36  wherein said alphavirus vector is semliki forest virus vector.  
     
     
         38 . A method according to  claim 33  or  claim 34  wherein said cell is an APC cell.  
     
     
         39 . A method according to  claim 38  wherein said APC cell is selected from the group consisting of a dendritic cell, a lymphocyte and a tumour cell.  
     
     
         40 . A method according to  claim 25  wherein the immune response is a CD8 +  T cell immune response.  
     
     
         41 . A method according to  claim 25  wherein said plurality of epitopes are from a self antigen.  
     
     
         42 . A method according to  claim 25  wherein said plurality of epitopes are from a tumour antigen.  
     
     
         43 . A method according to  claim 42  wherein the tumour is melanoma.  
     
     
         44 . A method according to  claim 25  wherein said plurality of epitopes are from a pathogen.  
     
     
         45 . A method according to  claim 44  wherein said pathogen is influenza virus.  
     
     
         46 . A method according to  claim 25  wherein said individual is a test animal and the epitopes are test epitopes.  
     
     
         47 . A method according to  claim 46  further comprising the step of determining the CTL response to each of the epitopes under test.  
     
     
         48 . A method according to  claim 46  wherein the test animal is a transgenic animal that provides an immune enviroment close to the human immune environment.  
     
     
         49 . A method according to  claim 48  wherein the test animal is an A2 transgenic mouse.  
     
     
         50 . A method of boosting an immune response to a plurality of epitopes in an individual, said individual having been previously exposed to at least one of said plurality of epitopes; said method comprising the steps of administering to the individual a composition comprising a plurality of peptides each peptide comprising one of said plurality of epitopes.  
     
     
         51 . A method according to  claim 50  wherein the individual has been previously exposed to at least one of said plurality of epitopes either naturally or by administering one or more peptides comprising one or more of said epitopes.  
     
     
         52 . A method according to  claim 51  wherein the immune response is a CD8 +  T cell immune response.  
     
     
         53 . A method according to  claim 51  wherein said plurality of epitopes are from a self antigen.  
     
     
         54 . A method according to  claim 51  wherein said plurality of epitopes are from a tumour antigen.  
     
     
         55 . A method according to  claim 54  wherein the tumour is melanoma.  
     
     
         56 . A method according to  claim 51  wherein said plurality of epitopes are from a pathogen.  
     
     
         57 . A method according to  claim 56  wherein said pathogen is influenza virus.  
     
     
         58 . A method according to  claim 50  or  claim 51  wherein the individual is a test animal and the epitopes are test epitopes.  
     
     
         59 . A method according to  claim 58  further comprising the step of determining the CTL response to each of the epitopes under test.  
     
     
         60 . A method according to  claim 58  or  claim 59  wherein the test animal is a transgenic animal that provides an immune environment close to the human immune environment.  
     
     
         61 . A method according to  claim 60  wherein the test animal is an A2 transgenic mouse.  
     
     
         62 . A method according to  claim 50  wherein each peptide is associated with an antigen presenting cell.  
     
     
         63 . A method according to  claim 62  wherein the antigen presenting cell is a dendritic cell.  
     
     
         64 . A method according to  claim 62  wherein the antigen presenting cell is a lymphocyte.  
     
     
         65 . A method of inducing an immune response to a plurality of epitopes in an individual, said method comprising the steps of administering to the individual a priming composition comprising one or more peptides comprising one or more of said plurality of epitopes and then administering a boosting composition which comprises a plurality peptides each encoding one of said plurality of epitopes.  
     
     
         66 . A method according to  claim 65  wherein the immune response is a CD8 +  T cell immune response.  
     
     
         67 . A method according to  claim 65  wherein said plurality of epitopes are from a self antigen.  
     
     
         68 . A method according to  claim 65  wherein said plurality of epitopes are from a tumour antigen.  
     
     
         69 . A method according to  claim 68  wherein the tumour is melanoma.  
     
     
         70 . A method according to  claim 69  wherein said plurality of epitopes are from a pathogen.  
     
     
         71 . A method according to  claim 70  wherein said pathogen is influenza virus.  
     
     
         72 . A method according to  claim 65  wherein the individual is a test animal and the epitopes are test epitopes.  
     
     
         73 . A method according to  claim 72  further comprising the step of determining the CTL response to each of the epitopes under test.  
     
     
         74 . A method according to  claim 72  or  claim 73  wherein the test animal is a transgenic animal that provides an immune environment close to the human immune environment.  
     
     
         75 . A method according to  claim 74  wherein the test animal is an A2 transgenic mouse.  
     
     
         76 . A method according to  claim 65  wherein each peptide is associated with an antigen presenting cell.  
     
     
         77 . A method according to  claim 76  wherein the antigen presenting cell is a dendritic cell.  
     
     
         78 . A method according to  claim 76  wherein the antigen presenting cell is a lymphocyte.  
     
     
         79 . A method of boosting an immune response to a plurality of epitopes in an individual, said individual having been previously exposed to at least one of said plurality of epitopes; said method comprising the steps of administering to the individual a composition comprising a plurality of individual vehicles each carrying a nucleic acid construct encoding one of said plurality of epitopes.  
     
     
         80 . A method according to  claim 79  wherein the individual has been previously exposed to at least one of said plurality of epitopes either naturally or by administering one or more vehicles carrying nucleic acid constructs encoding said epitopes, said vehicles being different to and non-reactive with any one of the plurality of individual vehicles used to boost the immune response.  
     
     
         81 . A method according to  claim 79  or  claim 80  wherein the individual is a test animal and the epitopes are test epitopes.  
     
     
         82 . A method according to  claim 81  further comprising the step of determining the CTL response to each of the epitopes under test.  
     
     
         83 . A method according to  claim 81  or  claim 82  wherein the test animal is a transgenic animal that provides an immune environment close to the human immune environment.  
     
     
         84 . A method according to  claim 79  wherein the immune response is a CD8 +  T cell immune response.  
     
     
         85 . A method according to  claim 79  wherein said plurality of epitopes are from a self antigen.  
     
     
         86 . A method according to  claim 79  wherein said plurality of epitopes are from a tumour antigen.  
     
     
         87 . A method according to  claim 86  wherein the tumour is melanoma.  
     
     
         88 . A method according to  claim 79  wherein said plurality of epitopes are from a pathogen.  
     
     
         89 . A method according to  claim 88  wherein said pathogen is influenza virus.  
     
     
         90 . A method according to  claim 83  wherein the test animal is an A2 transgenic mouse.  
     
     
         91 . A method according to  claim 79  wherein the vehicle is a nucleic acid expression vector.  
     
     
         92 . A method according to  claim 91  wherein the nucleic acid expression vector is a viral vector.  
     
     
         93 . A method according to  claim 92  wherein said viral vector is a semliki forest virus vector.  
     
     
         94 . A method of inducing an immune response to a plurality of epitopes in an individual, said method comprising the steps of administering to the individual a priming composition comprising a nucleic acid construct encoding at least one of said plurality of epitopes and then administering a boosting composition which comprises a plurality of nucleic acid constructs each encoding one of said plurality of epitopes.  
     
     
         95 . A method according to  claim 94  wherein the immune response is a CD8 +  T cell immune response.  
     
     
         96 . A method according to  claim 94  wherein said plurality of epitopes are from a self antigen.  
     
     
         97 . A method according to  claim 94  wherein said plurality of epitopes are from a tumour antigen.  
     
     
         98 . A method according to  claim 97  wherein the tumour is melanoma.  
     
     
         99 . A method according to  claim 94  wherein said plurality of epitopes are from a pathogen.  
     
     
         100 . A method according to  claim 99  wherein said pathogen is influenza virus.  
     
     
         101 . A method according to  claim 94  wherein the individual is a test animal and the epitopes are test epitopes.  
     
     
         102 . A method according to  claim 101  further comprising the step of determining the CTL response to each of the epitopes under test.  
     
     
         103 . A method according to  claim 101  or  claim 102  wherein the test animal is a transgenic animal that provides an immune environment close to the human immune environment.  
     
     
         104 . A method according to  claim 103  wherein the test animal is an A2 transgenic mouse.  
     
     
         105 . A method according to  claim 94  wherein the nucleic acid constructs are carried by a vehicle, said vehicle being a nucleic acid expression vector.  
     
     
         106 . A method according to  claim 105  wherein the nucleic acid expression vector is a viral vector and wherein the boosting composition comprises multiple viral vectors, said multiple viral vectors being different to and non-cross reactive with, the viral vector(s) used in the priming composition.  
     
     
         107 . A composition for boosting an immune response to one or more epitopes in an individual, said individual having been previously exposed to said one or more epitopes, said composition comprising a recombinant replication incompetent alphavirus vector comprising a heterologous nucleic acid sequence encoding one or more epitopes, and a pharmaceutically acceptable carrier.  
     
     
         108 . A composition according to  claim 107  wherein the recombinant replication incompetent alphavirus vector is a semiliki forest virus vector.  
     
     
         109 . A composition according to  claim 107  further comprising an adjuvant.  
     
     
         110 . A composition according to  claim 107  wherein said heterologous nucleic acid sequence further encodes a peptide adjuvant.  
     
     
         111 . A method of boosting an immune response to at least one epitope in an individual, said individual having been previously exposed to said at least one epitope; said method comprising administering to the individual a recombinant replication incompetent alphavirus vector comprising a heterologous nucleic acid sequence encoding said at least one epitope.  
     
     
         112 . A method according to  claim 111  wherein said alphavirus vector is a SFV vector.  
     
     
         113 . A method for inducing an immune response to at least one epitope in an individual; said method comprising the steps of 
 (a) administering to said individual a priming composition comprising said at least one epitope wherein said at least one epitope is optionally carried by a priming vehicle;    (b) administering to said individual a boosting composition comprising a recombinant replication incompetent alphavirus vector comprising a heterologous nucleic acid sequence encoding said at least one epitope; and wherein said alphavirus vector is non-cross reactive with the optional priming vehicle.    
     
     
         114 . A method according to  claim 113  wherein said alphavirus vector is a SFV vector.  
     
     
         115 . A method according to  claim 113  wherein the priming composition comprises a nucleic acid sequence, said nucleic acid sequence encoding said at least one epitope and wherein said nucleic acid sequence is optionally carried by a priming vehicle being a nucleic acid expression vector or a viral vector.  
     
     
         116 . A method according to  claim 113  wherein the priming composition comprises a peptide, polypeptide or protein comprising said at least one epitope.  
     
     
         117 . A method according to  claim 113  wherein the immune response is a CD8 +  T cell immune response.  
     
     
         118 . A method according to  claim 113  wherein said at least one epitope is from a self antigen.  
     
     
         119 . A method according to  claim 113  wherein said at least one epitope is from a tumour antigen.  
     
     
         120 . A method according to  claim 119  wherein the tumour is melanoma.  
     
     
         121 . A method according to  claim 113  wherein said plurality of epitopes are from a pathogen.  
     
     
         122 . A method according to  claim 121  wherein said pathogen is influenza virus.  
     
     
         123 . A prime-boost immunisation regimen for immunising an individual against at least one epitope, comprising 
 (a) a priming composition comprising a priming vehicle carrying said at least one epitope;    (b) a boosting composition comprising a boosting vehicle carrying said at least one epitope; wherein said boosting vehicle is an alphavirus vector and wherein said priming vehicle is different and non-reactive with said boosting vehicle.    
     
     
         124 . A prime-boost immunisation regimen according to  claim 123  wherein said priming composition comprises one or more peptides comprising said at least one epitope.  
     
     
         125 . A prime-boost immunisation regimen according to  claim 123  wherein said boosting composition comprises one or more peptides comprising said at least one epitope.  
     
     
         126 . A prime-boost immunisation regimen according to  claim 123  wherein said priming composition comprises one or more nucleic acid sequences encoding said at least one epitope.  
     
     
         127 . A prime-boost immunisation regiment according to  claim 123  wherein said boosting composition comprises one or more nucleic acid sequences encoding said at least one epitope.  
     
     
         128 . A prime-boost regimen according to  claim 123  wherein the priming vehicle is selected from the group consisting of a nucleic acid expression vector and an APC.  
     
     
         129 . A prime-boost regimen according to  claim 128  wherein the nucleic acid expression vector is a viral vector.  
     
     
         130 . A prime-boost regimen according to  claim 123  wherein the boosting vehicle is SFV.  
     
     
         131 . A prime-boost regimen according to  claim 123  wherein said priming composition and said boosting composition further comprise an adjuvant.  
     
     
         132 . A prime-boost regimen according to  claim 123  wherein said at least one epitope is from a self-antigen.  
     
     
         133 . A prime-boost regimen according to  claim 123  wherein said at least one epitope is from a tumour antigen.  
     
     
         134 . A prime-boost regimen according to  claim 123  wherein said tumour is melanoma.  
     
     
         135 . A prime-boost regimen according to  claim 123  wherein said at least one epitope is from a pathogen.  
     
     
         136 . A prime-boost regimen according to  claim 135  wherein said pathogen is influenza virus.  
     
     
         137 . A chimeric multimeric MHC structure that is capable of detecting specific CTLs expanded following vaccination of an individual or test animal with one or more epitopes, wherein said multimeric MHC structure comprises two or more human MHC molecules held together in a single structure by a binding member, said MHC molecules containing an altered α3 domain representing a murine α3 domain.  
     
     
         138 . A chimeric multimeric MHC structure according to  claim 137  wherein said altered α3 domain is a murine α3 domain.  
     
     
         139 . A chimeric multimeric MHC structure according to  claim 137  or  claim 138  further complexed with peptides displaying the epitopes used in the vaccination.  
     
     
         140 . A chimeric multimeric MHC structure according to any one of  claims 137  to  139  having four human MHC molecules.  
     
     
         141 . A chimeric multimeric MHC structure according to any one of  claims 137  to  140  wherein the chimeric MHC molecules are fusion proteins comprising human α1 and α2 domains and a murine α3 domain.