Means for detection and purification of CD8+ T lymphocyte populations specific to peptides presented in the context of HLA

Abstract

The invention concerns multimers developed from recombinant proteins analogues of MHC class I.

1 - 13 . (Canceled).  
     
     
         14 . Multimers built up from recombinant proteins analogues of class I MHC, characterized in that the proteins comprise at least one modification in the zone of interaction of a heavy chain with the CD8 co-receptor of T lymphocytes leading to a reduction, or even suppression of the affinity of the interaction between the heavy chain and CD8.  
     
     
         15 . Multimers according to  claim 14 , characterized in that the modification relates to the α3 domain of the heavy chain.  
     
     
         16 . Multimers according to  claim 14 , characterized in that the modification corresponds to a mutation in the α3 domain of at least one amino acid, with respect to the corresponding domain of a native heavy chain capable of binding to the said CD8co-receptor.  
     
     
         17 . Multimers according to  claim 14 , characterized in that the modification corresponds to chemical modification of at least one amino acid of the α3 domain of a heavy chain, with respect to the corresponding domain of a native heavy chain capable of binding to the said CD8 co-receptor.  
     
     
         18 . Multimers according to  claim 14 , characterized in that the modification corresponds to the deletion of at least one amino acid of the α3 domain of a heavy chain, with respect to the corresponding domain of a native heavy chain capable of binding to the said CD8 co-receptor.  
     
     
         19 . Multimers according to  claim 14 , characterized in that they are in the form of complexes with antigenic peptides.  
     
     
         20 . Multimers according to  claim 19 , characterized in that they are in the form of tetramers.  
     
     
         21 . Use of multimers according to  claim 19  for the purpose of detection and/or isolation of peptide-specific CD8+ T lymphocyte populations.  
     
     
         22 . Use according to  claim 21  in a process for cell screening, such as immunomagnetic screening.  
     
     
         23 . Method for the detection of peptide-specific CD8+ T lymphocyte populations from a polyclonal population, characterized in that it comprises: 
 bringing the polyclonal population into contact with multimers complexed with antigenic peptides according to  claim 19  under conditions which allow interaction between the modified class I MHC/peptide complexes and T lymphocyte receptors which have an affinity for the said complexes,    visualization of the lymphocyte populations which are bound to the said complexes.    
     
     
         24 . Method for isolation of peptide-specific CD8+ T lymphocyte populations from a polyclonal population, characterized in that it comprises: 
 bringing the polyclonal population into contact with magnetic beads on which are bound the peptide/class I CMH analogue complexes according to  claim 19  under conditions which allow interaction between the said complexes and T lymphocyte receptors which have an affinity for the said complexes, 
 recovery of the bound populations, the screening operation being repeated, if desired, and/or followed, where appropriate, by a stage  
 of in vitro amplification of the populations selected.  
   
     
     
         25 . Lymphocyte populations which have been selected and, where appropriate, amplified, characterized in that they are made up exclusively of T lymphocytes which are reactive towards the peptide of a complex with multimers according to  claim 19 .  
     
     
         26 . Pharmaceutical compositions which can be used, in particular, in immunotherapy, characterized in that they are built up from a lymphocyte population according to  claim 25  in combination with a pharmaceutically inert vehicle.