Identification, Optimization And Use Of Cryptic HLA-B7 Epitopes For Immunotherapy
a technology of hla-b7 and cryptic hlab7, which is applied in the field of peptide immunotherapy, can solve the problems of insufficient mhc i affinity, presence of primary anchor residues, and often necessary, and achieve the effect of enhancing affinity and immunogenicity of hla-b
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example 1
Affinity of Peptides
[0075]Eight peptides with the HLA-B*0702 specific anchor motifs, i.e. P2 and preferentially L / V at C-terminal position (Sidney et al., 1996) belonging to Hsp70 (Hsp70115, Hsp70137, Hsp70397), TERT (TERT4 and TERT444), and MAGE-A (MAGE-A121.1, MAGE-A121.2 and MAGE-A121.4) antigens were tested for binding to the HLA-B*0702 molecule. Only TERT4 bound to HLA-B*0702 with a high affinity, the remaining seven peptides were very weak or non binders (Table II). This demonstrates that the presence of anchor motifs is not sufficient to ensure a high binding affinity to HLA-B*0702. Given their low affinity, peptides Hsp70115, Hsp70137, Hsp70397, TERT444, MAGE-A121.1, MAGE-A121.2, MAGE-A121.4, are considered cryptic peptides.
TABLE IIHLA-B*0702 affinity of peptidesSEQ IDPeptideSequenceRANO:1Hsp70 115YPEEISSMVL>1011Hsp70 115A1APEEISSMVL>10122Hsp70 137 (10)YPVTNAVITV>10133Hsp70 397APLSLGLET>10144TERT4APRCRAVRSL 0.74155TERT444DPRRLVQLL>101TERT 444A1APRRLVQLL 1.456MAGE-A121.1EPV...
example 2
Immunogenicity of Selected Peptides
[0076]The low affinity Hsp137, Hsp115, Hsp397, TERT444 and the high affinity TERT4 peptides have been tested for their capacity to induce a specific CTL immune response in HLA-B*0702 transgenic mice. Only the high affinity TERT4 was immunogenic confirming that immunogenicity of peptides is strongly related to their affinity for HLA (FIG. 1).
example 3
Enhancement of Affinity of Low Affinity Peptides
[0077]Since all these cryptic peptides had favourable primary anchor motifs, enhancement of their affinity is a prerequisite for them to be immunogenic. It required the identification of unfavourable secondary anchor motifs and their substitution with favourable motifs. These substitutions should however preserve the conformation of the peptide segment that interacts with the TCR (position 4 to position 8). The interest was, therefore, focused on secondary anchor positions 1 and 3: aliphatic amino acids are favourable motifs at position 1 (Sidney, Southwood et al., 1996). However, peptides Hsp70115 and Hsp70137 that have a Y (tyrosine) at position 1 are non binders. Moreover, the substitution of the amino acid at position 1 by an A (alanine) that is also favourable at this position. (Parker et al, 1994) enhances the affinity of the TERT444 but not of the Hsp70115 and the MAGE-A121.1 peptides (Table II). This indicates that the presence...
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