Plasmids with immunological action

Abstract

Recombinant plasmids usable for the transfection of eukaryotic and prokaryotic cells are described; such plasmids have a length comprised between 7 and 12 kbases and comprise a sequence encoding the heavy chain of an immunoglobulin; in particular, they may be used: in a process of transfection of prokaryotic or eukaryotic cells (ex vivo) which can be inoculated into higher organisms in order to induce a prophylactic or therapeutic immune response; in a protocol of direct inoculation (in vivo) in higher organisms in genic immunization methodologies with the aim of evoking prophylactic or therapeutic immune responses.

1 .- 31 . (canceled) 
     
     
         32 . A recombinant plasmid having a length of between 7 and 12 kilobases and comprising a sequence encoding the heavy chain of immunoglobulin, wherein the plasmid contains at least one sequence encoding an antigenic epitope and the plasmid does not express resistance to a betalactamic antibiotic. 
     
     
         33 . The plasmid according to  claim 32 , wherein said betalactamic antibiotic is ampicillin. 
     
     
         34 . The plasmid according to  claim 32 , wherein it has a length of between 8 and 12 kilobases. 
     
     
         35 . The plasmid according to  claim 32 , wherein it has a length of between 9 and 11 kilobases. 
     
     
         36 . The plasmid according to  claim 32 , wherein it has a length of between 9 and 10 kilobases. 
     
     
         37 . The plasmid according to  claim 32 , wherein said heavy chain of the immunoglobulin is chimeric. 
     
     
         38 . The plasmid according to  claim 37 , wherein said heavy chain of the immunoglobulin comprises a murine variable region and a human constant region. 
     
     
         39 . The plasmid according to  claim 38 , wherein said murine variable region is V H 62. 
     
     
         40 . The plasmid according to  claim 39 , wherein said murine variable region V H 62 is cloned from a mouse hybridoma secreting a monoclonal antibody with anti-tyroglobulin activity. 
     
     
         41 . The plasmid according to  claim 38 , wherein said human constant region is Igγ 1 . 
     
     
         42 . The plasmid according to  claim 41 , wherein said human constant region Igγ 1  is cloned from the vector pNγ 1 . 
     
     
         43 . The plasmid according to  claim 32 , wherein it comprises a promoter for lymphocytic cells. 
     
     
         44 . The plasmid according to  claim 43 , wherein said promoter for lymphocytic cells is around 50 bp. 
     
     
         45 . The plasmid according to  claim 43 , wherein said promoter for lymphocytic cells is a viral promoter. 
     
     
         46 . The plasmid according to  claim 45 , wherein said viral promoter is around 742 bp. 
     
     
         47 . The plasmid according to  claim 32 , wherein it comprises the polyadenylation sequence AATAAA. 
     
     
         48 . The plasmid according to  claim 32 , wherein it comprises an  Escherichia coli  bacterial origin of replication. 
     
     
         49 . The plasmid according to  claim 32 , wherein it expresses resistance to neomycin and it comprises the origin of replication pBR322. 
     
     
         50 . The plasmid according to  claim 32 , wherein it does not comprise the replication origin SV40. 
     
     
         51 . The plasmid according to  claim 32 , wherein said antigenic epitope is selected from the group consisting of p540 (ILAKFLHWL; SEQ ID NO: 3), p572 (RLFFYRKSV; SEQ ID NO: 4), pY572 (YLFFYRKSV; SEQ ID NO: 5), p865 (RLVDDFLLV; SEQ ID NO: 6) and pNP (ASNENMETM; SEQ ID NO: 7). 
     
     
         52 . A recombinant plasmid having a sequence at least 90% homologous to SEQ ID NO: 1. 
     
     
         53 . The recombinant plasmid according to  claim 52 , wherein it has a sequence at least 95% homologous to SEQ ID NO: 1. 
     
     
         54 . The recombinant plasmid according to  claim 52 , wherein it has the sequence set forth in SEQ ID NO: 1. 
     
     
         55 . A recombinant plasmid having a sequence at least 90% homologous to SEQ ID NO: 2. 
     
     
         56 . The recombinant plasmid according to  claim 55 , wherein it has a sequence at least 95% homologous to SEQ ID NO: 2. 
     
     
         57 . The recombinant plasmid according to  claim 50 , wherein it has the sequence set forth in SEQ ID NO: 2. 
     
     
         58 . A formulation containing at least one plasmid according to  claim 32  together with pharmaceutically acceptable excipients and coadjuvants. 
     
     
         59 . A method for inducing an immunoresponse in a human or animal organism comprising the transfection of prokaryotic or eukaryotic cells ex vivo by means of a plasmid according to  claim 32  and the subsequent inoculation of said prokaryotic or eukaryotic cells into said human or animal organism. 
     
     
         60 . The method according to  claim 59 , wherein the cells to be transfected belong to the family of lymphocytes. 
     
     
         61 . The method according to  claim 60 , wherein said cells are taken from the peripheral vessels of said human or animal organism. 
     
     
         62 . The method according to  claim 59 , wherein said plasmid has the sequence SEQ ID NO: 1. 
     
     
         63 . The method according to  claim 59 , wherein said plasmid has a sequence at least 95% homologous with respect to SEQ ID NO: 1. 
     
     
         64 . A method for inducing an immunoresponse in a human or animal organism comprising the inoculation in vivo into said human or animal organism of a plasmid according to  claim 32 . 
     
     
         65 . The method according to  claim 64 , wherein said plasmid has the sequence SEQ ID NO: 2. 
     
     
         66 . The method according to  claim 64 , wherein said plasmid has a sequence at least 95% homologous to SEQ ID NO: 2. 
     
     
         67 . The method according to  claim 59 , wherein the inoculation of the plasmid or of the prokaryotic or eukaryotic cells into said human or animal organism is carried out by means of injective or transmucosal administration.